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Sequencing and Phylogenetic Analysis of the RPB1 Gene of Nosema sp. PA YAN Wei1, SHEN Zhong-yuan, TANG Xu-dong, XU Li, LI Qian-long, XIAO Sheng-yan, YUE Ya-jie, FU Xu-liang Scientia Agricultura Sinica    2014, 47 (23): 4736-4744.   DOI: 10.3864/j.issn.0578-1752.2014.23.018 Abstract （472）   HTML （2）    PDF （5779KB）（575）       Knowledge map    Save 【Objective】 The objective of this study is to clarify the taxonomic status of Nosema sp. PA and provide a new foundation for the further study of its biological function by cloning the RPB1 (largest subunit of RNA polymeraseII) gene of Nosema sp. PA and analyzing the gene sequence by bioinformatics methods. 【Method】 Six pairs of homologous primers were designed by Primer Premier 5.0 software for the RPB1 gene of Nosema sp. PA based on the RPB1 gene of Nosema bombycis. Partial sequence of the RPB1 gene of Nosema sp. PA was cloned by PCR amplification. Then, bioinformatics analysis on the RPB1 gene of Nosema sp. PA and its encoding protein were conducted by bio-softs as GSDS, SMART, DNAstar and MEGA4.1. 【Result】 Partial sequence of the RPB1 gene of Nosema sp. PA was cloned by PCR amplification (GenBank accession number KJ728831). The partial sequence of the RPB1 gene of Nosema sp. PA had 2 933 nucleotides which contained an ORF with 2 922 bp encoding a polypeptide of 974 amino acids with a molecular weight of 109.38 kD and an isoelectric point of 7.087. The structure of the partial sequence of the RPB1 gene was a single exon. The encoded protein contained four domains: RPOLA_N, RNA_pol_Rpb1_4, RNA_pol_Rpb1_5 and RNA_pol_Rpb1_6. RPOLA_N domain is a very important domain both in prokaryotes and eukaryotes among the four domains. The encoded protein contained four main secondary structures: α-helix, random coil, β-turn and extended strand. The proportion of α-helix and random coil was quite high. Extended strand was mainly located between the α-helix and random coil. Sequence comparison and phylogenetic analysis showed that the encoded protein of Nosema sp. PA was 81.9%-99.6% identity and 0.004-0.049 divergence with those of N. bombycis, N. trichoplusiae, Nosema sp. CPP, N. fumiferanae, N. disstriae, N. tyriae and N. granulosis. Nosema sp. PA and other seven kinds of “true’’ Nosema species above-mentioned clustered in the same clade. The encoded protein was 99.6% identical to N. bombycis and had a close relationship with N. bombycis. 【Conclusion】The partial sequence of the RPB1 gene of Nosema sp. PA is successfully cloned. The results confirmed that Nosema sp. PA is a member of Nosema species in the aspect of molecular biology. Reference | Related Articles | Metrics

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Effect of Different Amounts of Compost Fertilizer on Yields and Nutritive Values of Whole Corn Plant Silage LI Meng-Meng, ZHANG Gui-Guo, YANG Zai-Bin, YANG Wei-Ren Scientia Agricultura Sinica    2013, 46 (11): 2337-2344.   DOI: 10.3864/j.issn.0578-1752.2013.11.018 Abstract （626）      PDF （585KB）（697）       Knowledge map    Save 【Objective】The objective of the present study was to investigate the effects of fertilizing compost and chemical fertilizer on feed production potential of the whole corn plant silage, and thus providing a theoretical support for appropriate application of compost in corn silage cultivation. 【Method】 A field trial was conducted with a single-factor randomized block design. Three fertilizer treatments were different amounts of compost at 20, 40, and 60 t•hm-2, respectively. Chemical fertilizer with urea 400 kg•hm-2, superphosphate 450 kg•hm-2 and potassium sulfate (K2SO4) 300 kg•hm-2 was used as control. Biomass, dry matter (DM) yield, nutrients content and ruminal degradability were measured to elucidate the effect of compost amount on yields of DM, nutrients and degradable nutrients per unit land of the silage corn. 【Result】 Compared with control, the contents of crude protein (CP), ether extract (EE) and gross energy (GE) of whole plant corn silage were significantly increased (P＜0.05) with the increased amount of compost. The content of crude fiber (CF), however, was notably decreased (P＜0.05), and there was no significant difference (P＞0.05) in carbohydrate content. Meanwhile, ruminal degradabilities of CP, EE, carbohydrate, and GE of corn silage were enhanced (P＜0.05) with increased fertilizer amount of compost. Regardless of fertilizer amount of compost, yields of biomass, nutrients, ruminally degradable nutrients, and relative feed value (RFV) of corn silage were higher than that of control (P＜0.05). 【Conclusion】 The results showed that compost administration can improve corn silage biomass and feeding values compared with administration of chemical fertilizer. Under the present experimental conditions, the optimal fertilizing amount of compost was 40 t•hm-2. Reference | Related Articles | Metrics

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Optimization of Reaction Conditions for Cello-Oligosaccharides Production from Wheat Straw by β-Glucanase Hydrolysis LIU Sha-Sha, LI Jing-Mei, SHI Bo, LIANG Ping, LI Chao Scientia Agricultura Sinica    2013, 46 (11): 2345-2352.   DOI: 10.3864/j.issn.0578-1752.2013.11.019 Abstract （623）      PDF （598KB）（710）       Knowledge map    Save 【Objective】 The aim of this study was to explore the optimum conditions of wheat straw degradation by β-glucanase.【Method】The mixed-acid hydrolysis and chromatography were selected for the separation process, in which the cello- oligosaccharides with degrees of polymerization from 2 to 5 were obtained. The single factor experiments for optimal conditions were temperature, pH, E/S concentration of wheat straw and the hydrolysis time. The optimal conditions for hydrolysis of wheat straw were obtained through analyzing the yield of cellobiose and the conversion rate of wheat straw.【Result】The experimental results showed that by activated carbon column chromatography, the standard cello-oligosaccharides were obtained and the optimum conditions for wheat straw hydrolysis were as follows: the reaction temperature was 50℃, pH was 5.5, E/S was 0.4, concentration of wheat straw was 2%, and hydrolysis time was 10 h. Using the optimum conditions, the cellobiose yield was 148.15 mg•g-1 wheat straw and the conversion rate of wheat straw was 55.57%. 【Conclusion】 The yield of cellobiose and the conversion rate of wheat straw were both increased under the optimum conditions. It was found that the chromatographic columns composed of charcoal could separate well cello-oligosaccharides with degrees of polymerization from 2 to 5. Reference | Related Articles | Metrics

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Identification of Silkworm Pupa-Specific Gene BmCP283 and Its Promoter CHENG Dao-Jun, TANG Lin, MENG Meng, KANG Li-Xia, WANG Yong-Hu, PENG Jian, MA San-Yuan, XIA Qing-You Scientia Agricultura Sinica    2013, 46 (11): 2353-2362.   DOI: 10.3864/j.issn.0578-1752.2013.11.020 Abstract （681）      PDF （771KB）（991）       Knowledge map    Save 【Objective】The objective of this study is to identify genes that are specifically expressed during silkworm pupa-adult transition, clone their promoters and provide supports for controlling artificially pupa-adult transition and develope pupal bioreactor in silkworm (Bombyx mori).【Method】Silkworm pupa-specific genes were identified through analysis of microarray data for gene expression during silkworm metamorphosis and confirmed by RT-PCR experiments. The promoters of silkworm pupa-specific genes were cloned by PCR experiments and their driving activities and pupa-specificity were detected by transgenic technology.【Result】Cuticular protein gene BmCP283 was found to be specifically expressed at pupal stage in silkworm and was regarded as a pupa-specific gene. The potential promoter with a length of 2 004 bp on the upstream of the translational initial site of BmCP283 was cloned. Transgenic experiments confirmed that the promoter of BmCP283 could drive the red fluorescent gene dsRed to be specifically expressed at late pupal stages, which was similar to the expression profile of endogenous BmCP283. 【Conclusion】Both the expression pattern of BmCP283 and the activities of its promoter is pupa-specific in silkworm. Reference | Related Articles | Metrics

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Molecular Cloning and Preliminary Functional Analysis of Domains of Duck Retinoic Acid Inducible Gene I CHEN Yang, HUANG Zheng-Yang, ZHANG Yang, LI Xin-Yu, ZHEN Ting, WU Ning-Zhao, XU Qi, CHEN Guo-Hong Scientia Agricultura Sinica    2013, 46 (10): 2094-2102.   DOI: 10.3864/j.issn.0578-1752.2013.10.015 Abstract （555）      PDF （771KB）（797）       Knowledge map    Save 【Objective】Duck RIG-I (duRIG-I) gene was cloned and the functions of its different domains were analyzed preliminarily. 【Method】 The CDS of duRIG-I gene was cloned on the basis of the sequence submitted to GenBank with RT-PCR and was analyzed by bioinformatics. The eukaryotic expression vectors of N-terminal, C-terminal and whole-length of duRIG-I gene with 6*his tags were constructed to transfect DF1, and then the transcription and expression of the three recombinant plasmids in cells were detected via RT-PCR and indirect immunofluorescent assay, respectively. Meanwhile, the expressions of chicken IFN-β, Mx1 and PKR mRNA were detected by real-time PCR.【Result】The whole-length of duRIG-I CDS was 2 802 bp encoding 933 amino acids. All the recombinant protein of duRIG-I could express normally in DF1. The results of RT-qPCR indicated that CARDs significantly up-regulated the mRNA level of IFN-β, Mx1 and PKR genes.【Conclusion】The various domain fragments of duRIG-I express normally in DF1. The N-terminal of duRIG-I plays a vital role in regulating the expression of downstream genes of RLR antiviral signal pathway. Reference | Related Articles | Metrics Cited: Baidu(1)

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Expression and Localization of Claudin-11 in Adult Alpaca Ovary GENG Jian-Jun, GUO Qing-Yun, PANG Ya-Miao, BAI Jun-Ming, HE Jun-Ping Scientia Agricultura Sinica    2013, 46 (10): 2103-2109.   DOI: 10.3864/j.issn.0578-1752.2013.10.016 Abstract （533）      PDF （763KB）（639）       Knowledge map    Save 【Objective】To explore the mechanism of Claudin-11 in promoting the follicular development and regulating the ovulation, an experiment was carried out to investigate the expression and localization of Claudin-11 in adult alpaca ovary. 【Method】 Using the adult female alpaca as the object of study, the expression of Claudin-11 in the ovary was analyzed by RT-PCR and real-time PCR. Immunohistochemistry and western blotting were used to explore the localization of Claudin-11 in alpaca ovary.【Result】The expression of Claudin-11 in ovarian primary follicle was weaker than in the secondary follicle. Polyclonal antibody happened specific immune response with the crude extract from the ovarian. The expression of Claudin-11 in ovarian primary follicle and secondary follicle was weaker than that in flat or cube follicle cell cytoplasm. Claudin-11 was mainly located in the granulosa cell of the secondary follicle endothecium.【Conclusion】The results showed that Claudin-11 expressed in adult alpaca ovary, and there was a specific cellular localization. Reference | Related Articles | Metrics

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Optimization of Incubation of Sperm and Lentivirus and Transgenic Pigs Production CAI Wei-Guang, XI Qian-Yun, XIAO Min, GUAN Li-Zeng, MENG Fan-Ming, SHU Gang, ZHU Xiao-Tong, JIANG Qing-Yan, WU Zhen-Fang, ZHANG Yong-Liang Scientia Agricultura Sinica    2013, 46 (9): 1903-1914.   DOI: 10.3864/j.issn.0578-1752.2013.09.018 Abstract （792）      PDF （692KB）（527）       Knowledge map    Save 【Objective】A new method was developed for producing transgenic pig using incubation of lentivirus and sperm. 【Method】Effects of sperm number, amount of lentivirus, incubation time and incubation temperature on sperm activity were assessed by the orthogonality design. PCR and southern blotting were used to analyze the transgene.【Result】The optimized incubation was incubation of 100 µL lentivirus (5×105cfu) with 1.0×107 sperms per mL for 30 min at 17℃. Using this procedure sperms were artificially inseminated into sows. PCR was used to detect the offspring. Fourteen of 49 (14/49) piglets were PCR-positive. Four of 8 PCR-positive pitlets showed foreign gene intergration revealed by using southern blotting.【Conclusion】Incubation indexes of lentiviruse and sperm optimized in this study are that 100 µL lentivirus (5×105cfu) with 1.0×107 sperms per mL for 30 min at 17℃, and transgenic pigs were produced by using this procedure. Foreign gene intergration were revealed. Reference | Related Articles | Metrics Cited: Baidu(1)

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Study on the Relationship Between Polymorphism of RERG Gene and Growth Traits of Three Goats Breeds in Guizhou CHEN Zhi, LUO Wei-Xing, LIU Ruo-Yu, CAI Hui-Fen, CHENG Jie, SONG Tao-Wei, ZHANG Yi-Yu Scientia Agricultura Sinica    2013, 46 (9): 1915-1922.   DOI: 10.3864/j.issn.0578-1752.2013.09.019 Abstract （655）      PDF （637KB）（595）       Knowledge map    Save 【Objective】This study aimed at researching polymorphism of the second, third and fourth exons of ras-related and estrogen-regulated growth inhibitor (RERG) gene in three indigenous Guizhou goats (Qianbei Ma goat, Guizhou White goat and Guizhou Black goat) and its association with growth traits was analyzed. 【Method】PCR-SSCP and PCR-direct sequencing were applied to detect the polymorphism sites and the general linear model was used to analyze its association with growth traits.【Result】Four consistent SNPs sites were found in the fourth exon of RERG gene in such three breeds. They are 56 bp C→G, 826 bp A→G, 1 434 bp T→C and 1 798 bp A→T. The following fixed effects model was employed in analyzing the growth traits in goat. The body height was significantly higher in individuals with genotype AB than AA and BB and genotype EF than EE and FF (P＜0.01). A significant difference was found in body weight between genotype CD and CC and DD (P＜0.05). The 1 798 bp (A/T) site did not appear significant difference. 【Conclusion】The 56 bp (C/G), 826 bp (A/G) and 1 434 bp (T/C) sites will provide a study basis between RERG genes and growth traits as a well as certain theoretical foundation for better feeding and genetic resources of indigenous goats. Reference | Related Articles | Metrics Cited: Baidu(3)

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Study on Fetal Skin Hair Follicle Development and Morphology of China Super-Fine Merino (Gansu Type) WU Yu-Yu, YUE Yao-Jing, GUO Ting-Ting, WANG Tian-Xiang, GUO Jian, LI Gui-Ying, HAN Ji-Long, YANG Min, LIU Jian-Bin, SUN Xiao-Ping, LI Fan-Wen, HE Yu-Qin, YANG Bo-Hui Scientia Agricultura Sinica    2013, 46 (9): 1923-1931.   DOI: 10.3864/j.issn.0578-1752.2013.09.020 Abstract （795）      PDF （732KB）（789）       Knowledge map    Save 【Objective】This study aims to research the organizational structure of China Super-Fine Merino hair follicles and hair follicle morphogenesis process, which will lay a histological basis for molecular regulation mechanism of Super-fine Merino hair follicle development.【Method】Tissue frozen section was conducted by embryonic skin of China Super-Fine Merino and the results were taken photography under a microscope.【Result】Results showed that the follicle structure of Super-fine Merino sheep consists of connective tissue sheath, outer root sheath, inner root sheath, hair shaft and hair bulb sections, as other mammals. Primary hair follicle occurred on 87th day of fetal period, vesicle structure of the secondary follicle could be found at the bottom of the primary hair follicle. At gestational 138 d, the primary basic hair follicle was mature. From gestational 87 d to 147 d, the density of primary follicle was gradually decreased and the density of secondary follicle was increased as the gestational age growing, meanwhile the density of primary follicle reached its maximum value (232.8±12.44) (per mm2) at gestational 117 d. The gestational Secondary Follicles to Primary Follicles ratio (S/P ratio) reached the maximum (9.96) at gestational age 126 d.【Conclusion】Primary follicles began to occur at 84 d of gestational age, and secondary hair follicle began to occur at 87 d of gestational age, gestational age 102 d in the original the secondary follicles neck bulge began regeneration to secondary-derived follicles, most of primary follicles and some of secondary hair follicle had matured. The differentiation of secondary hair follicle is a key factor which affected the density of hair follicle; it can effectively increase the density of hair follicle and improve the quality of the wool. Reference | Related Articles | Metrics Cited: Baidu(7)

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Screening and Analysis of Dipeptidyl Peptidase Ⅳ from Microbial Metagenomic Library in the Rumen of Dairy Cows ZHAO Jing-Wen, WANG Jia-Qi, ZHAO Sheng-Guo, SUN Peng, BU Deng-Pan, HU Xiao-Li, LU Yu-Fei, WANG Dan-Dan, JIN Di Scientia Agricultura Sinica    2013, 46 (8): 1687-1693.   DOI: 10.3864/j.issn.0578-1752.2013.08.019 Abstract （688）      PDF （580KB）（670）       Knowledge map    Save 【Objective】The purpose of this study was to reveal the characteristics of dipeptidyl peptidase Ⅳ (DPP-Ⅳ) sequences and enzymatic activity during the process of protein degradation in the rumen of dairy cows.【Method】The dpp-Ⅳ degenerate primers were used to screen rumen microbial Fosmid library including 17664 clones. The plasmids were extracted from the positive clones and digested by Hind Ⅲ. The dpp-Ⅳ sequences were obtained by PCR, cloning and sequencing. The Fosmid end sequences and DPP-Ⅳ sequences of the positive clones were analyzed by BLASTX and BLASTP, respectively. The peptidase activity from the positive clones was measured using Gly-Pro-pNA as a substrate.【Result】Ten positive clones named DP1-DP10 containing dpp-Ⅳ fragment were obtained. 78% of the Fosmid end sequences could match with the known genes (similarity 44%-94%). DPP-Ⅳ sequences contained N-conservative region (DWVYEEE) and C-catalytic domain (GWSYGG). DPP-Ⅳ sequences matched to Cyclobacterium marinum (43%), Capnocytophaga sp. (63%), Prevotella ruminicola 23 (66%) and Solitalea canadensis (50%). The activity of DP7 peptidase is the highest (6.88 U•mg-1).【Conclusion】Ten positive clones obtained from rumen microbial Fosmid library had different sequence characteristics and peptidase activity. This study implies that the substitution of some DPP-Ⅳ AA may affect the enzyme activity. Reference | Related Articles | Metrics Cited: Baidu(1)

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Advances in Study on Bio-Energy Utilization of Stem Cell Wall Components in Alfalfa (Medicago sativa L.) WANG Xiao-Juan, ZHANG Shu-Zhen, LIN Shuang-Shuang, DENG Zhi-Gang, JIN Liang Scientia Agricultura Sinica    2013, 46 (8): 1694-1705.   DOI: 10.3864/j.issn.0578-1752.2013.08.020 Abstract （633）      PDF （728KB）（970）       Knowledge map    Save Alfalfa (Medicago sativa L.) has been recognized as one of the potential biofuel crops with high biomass and wild adaptability. Most importantly, the stems of alfalfa would be processed to produce ethanol and the leaves could be served separately as a livestock feed. In this paper, the recent advances of the studies and utilization of alfalfa as energy plant were reviewed, which including germplasm evaluations of biofuel traits, development of stem cell wall, biosynthetic pathways and genetic control of cell wall components (cellulose, hemi-cellulose and lignin), and the effects of cultivation management system to ethanol production. The development of stem cell wall and genetic regulation of lignin in alfalfa were discussed. Furthermore, the perspective utilization of alfalfa as biofuel crop was also proposed. Reference | Related Articles | Metrics Cited: Baidu(8)

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Molecular Cloning and Expression Profiles Analysis of Chemosensory Protein Genes Family in the Chinese Honeybee (Apis cerana cerana) NI Cui-Xia, ZHANG Lin-Ya, LI Hong-Liang, SHANG Han-Wu Scientia Agricultura Sinica    2013, 46 (8): 1706-1715.   DOI: 10.3864/j.issn.0578-1752.2013.08.021 Abstract （657）      PDF （739KB）（912）       Knowledge map    Save 【Objective】 The objective of this study is to clone the whole genes family of chemosensory protein in the Chinese honeybee (Apis cerana cerana) (Ac-CSPs) and analyze their expression of mRNA in different tissues and different development time-points. This may provide a fundamental evidence for the future study of the physiological function of Ac-CSPs. 【Method】 Six full-length of CSP genes of A. cerana cerana was cloned by using reverse transcription-polymerase chain reaction (RT-PCR). The expression patterns at development phases and in different sensory organs of the six Ac-CSPs were analyzed by real-time PCR. 【Result】 Four full-length of Ac-CSP genes (Ac-CSP2, Ac-CSP4, Ac-CSP5 and Ac-CSP6) were cloned in which Ac-CSP2 was 354 bp in full length that encoded a polypeptide containing 117 amino acids with prediction of MW (molecular weight) of 13.01 kD and pI of 8.86, respectively. Ac-CSP4 was 387 bp in full length that encoded a polypeptide containing 128 amino acids with prediction of MW of 14.61 kD and pI of 4.53, respectively. Ac-CSP5 was 315 bp in full length that encoded a polypeptide containing 104 amino acids with prediction of MW of 12.46 kD and pI of 9.60, respectively. Ac-CSP6 was 378 bp in full length that encoded a polypeptide containing 125 amino acids with prediction of MW of 14.63 kD and pI of 8.71, respectively, and all of the genes with four conserved cysteine residues. The six deduced Ac-CSPs sequences showed high sequence similarity (95%-99%) as compared with CSPs (Am-CSPs) sequences of A. mellifera. The genes of Ac-CSP1, 2, 3, 4 were not expressed in eggs, larvae and pupae, with the Ac-CSP1, 2, 4 expressed at highest level in antennae and Ac-CSP3 was strongly expressed in wings. The expression of Ac-CSP5 was not significantly different at all developmental stages, while Ac-CSP6 only highly expressed in the pupae. 【Conclusion】 The alignment is very similar of Ac-CSPs and Am-CSPs and the expression patterns are similar but there is a part of difference in this research, which provides useful information for the study of function of CSPs in the future. Reference | Related Articles | Metrics Cited: Baidu(4)

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Construction of the Recombinant Adenovirus of FABP4 Gene of Qinchuan Cattle WEI Sheng-Juan, ZAN Lin-Sen, WANG Hong-Bao, CHENG Gong, JI Shu-Han, WANG Hong, FU Chang-Zhen, JIANG Bi-Jie Scientia Agricultura Sinica    2013, 46 (7): 1434-1440.   DOI: 10.3864/j.issn.0578-1752.2013.07.014 Abstract （797）      PDF （791KB）（448）       Knowledge map    Save 【Objective】The aim of this study was to construct the recombinant adenovirus vector with FABP4 gene of Chinese Qinchuan cattle, so as to provide a basis for studying FABP4 gene functions and mechanisms at cell level.【Method】The primers were designed according to the FABP4 mRNA sequence in GenBank, and the gene was cloned by RT-PCR and then sequenced. The bovine gene fragments containing both FABP4 gene and restriction enzyme were inserted into a shuttle vector pAdTrack-CMV to construct the recombinant shuttle vector pAdTrack-CMV-FABP4. After identifying by digestion and sequencing, pAdTrack-CMV-FABP4 plasmid was linearized by PmeⅠ, and then it was transformed into E. coli BJ5183 competent cells containing backbone vector pAdeasy-1 to obtain recombinant vector by homologous recombination. Then the positive plasmid was linearized by PacⅠ, and transfected into HEK 293A cells for virus packing, amplification and titer testing by TCID50.【Result】The results of enzyme digestion, sequencing and regular PCR detection showed that the recombinant overexpression vector containing FABP4 CDS region was successfully constructed. The infectious titer of the virus AD-FABP4 was 1.58×109 PFU•mL-1.【Conclusion】In this experiment, the recombinant adenovirus vector carrying FABP4 gene was constructed successfully and high titer adenovirus was acquired. Reference | Related Articles | Metrics Cited: Baidu(2)

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miR-133a Targets BIRC5 to Regulate Its Gene Expression in Chicken WANG Xing-Guo, SHAO Fang, GONG Dao-Qing, LU Xiang-Yun, GU Zhi-Liang Scientia Agricultura Sinica    2013, 46 (7): 1441-1447.   DOI: 10.3864/j.issn.0578-1752.2013.07.015 Abstract （540）      PDF （557KB）（605）       Knowledge map    Save 【Objective】This study was conducted to investigate the expression pattern of miR-133a in various chicken tissues, and find whether baculoviral IAP repeat containing 5 (BIRC5) is the target gene of miR-133a.【Method】The bioinformatics methods were used to predict the target genes of miR-133a. Real-time PCR was used to detect the expression patterns of miR-133a of various tissues in chicken. Dual-luciferase reporter assays and site mutation assays were used to verify whether BIRC5 is a target gene of miR-133a. 【Result】Two hundred and eighty-seven genes of 11891 genes from 3′UTR database were predicted as target genes of miR-133a. The expression patterns of miR-133a in various chicken tissues showed that miR-133a was highly expressed in muscle especially in skeletal muscle. Reporter assays showed that BIRC5 was the target gene of miR-133a and the site mutation assays validated the target site of miR-133a in BIRC5. 【Conclusion】 miR-133a is a miRNA related with skeletal muscle development of chicken, and BIRC5 is a bona fide target gene in chicken. Reference | Related Articles | Metrics Cited: Baidu(1)

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Proteome Comparison of Honeybee (Apis mellifera ligustica) Worker Venom Between Collected from Venom Glands and Electrical Stimulated LI Rong-Li, ZHANG Lan, HAN Bin, FANG Yu, FENG Mao, ZHOU Tian-E, LI Jian-Ke Scientia Agricultura Sinica    2013, 46 (7): 1448-1462.   DOI: 10.3864/j.issn.0578-1752.2013.07.016 Abstract （603）      PDF （940KB）（1554）       Knowledge map    Save 【Objective】The objective of this study is to investigate bee venom composition and difference by comparison of bee venom collected from venom glands and electrical stimulated of Italian bees (Apis mellifera ligustica), and to provide a theoretical basis for pharmaceutical application.【Method】The proteome between bee venom collected directly from the venom glands and artificially electrical stimulation were compared using gel-based (one-dimensional gel electrophoresis, 1-DE, two-dimensional gel electrophoresis, 2-DE) and gel-free proteomics approaches, mass spectrometry, and bioinformatics.【Result】In the gland venom, 19, 11 and 14 proteins were identified by 1-DE, 2-DE and shotgun analysis, respectively, which correspond to 30 nonredundant proteins. They were mainly classified as bee venom toxins (50%), antioxidant systems, protein folding and molecular transporters (50%). In construct, in electrical stimulated venom, 12, 3 and 7 proteins were identified, respectively, by 1-DE, 2-DE and shotgun analysis. They were classified into 14 nonredundant proteins, which the major components were venom toxins (93%). Notably, phospholipase A2-like protein was identified for the first time in two forms of bee venom and peptidyl-prolyl cis-trans isomerase was identified only in the gland venom for the first time. The abundance of apamin preproprotein and secapin in the gland venom were significantly higher than those in the electrical stimulated venom. However, phospholipase A-2, venom dipeptidyl peptidase IV precursor, venom allergen acid phosphatase and mast cell degranulating peptide had higher abundance in the electrical stimulated venom than those in the gland venom.【Conclusion】 The venom collected from venom glands contains more protein species, but the abundance of the toxin proteins in electrical stimulated venom are no less than the venom collected from the glands. As the pharmacological components are mainly contained in the toxin proteins, the electrical stimulated venom is a convenient and effective way for utilization. The identified new proteins significantly extend the knowledge of bee venom composition. The result may provide a theoretical and practical basis for future rational use of the honeybee venom. Reference | Related Articles | Metrics Cited: Baidu(6)

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Effect of FSH on the mRNA Expression of Cyclin D1 and Cyclin E1 in the Cultured Immature Boar Sertoli Cell ZHANG Jiao-Jiao, WANG Yi, ZHANG Hui-Qiong, SUN Si, SUN Yan, WANG Xian-Zhong, ZHANG Jia-Hua Scientia Agricultura Sinica    2013, 46 (6): 1237-1246.   DOI: 10.3864/j.issn.0578-1752.2013.06.018 Abstract （641）      PDF （556KB）（654）       Knowledge map    Save 【Objective】 The objective of the study is to identify whether or how FSH regulates the mRNA expression of cyclin D1 and cyclin E1 in the cultured immature boar Sertoli cell. 【Method】 In the study, the cultured immature boar Sertoli cells were used as materials, and real-time PCR was used to detect the mRNA expression of cyclin D1 and cyclin E1. 【Result】 Different doses of FSH (0-100 ng•mL-1) increased the mRNA expression of cyclin D1 and cyclin E1 (P＜0.05), which were peaking at 50 ng•mL-1 (P＜0.05). In addition, FSH (50 ng•mL-1) increased the mRNA expression of cyclin D1 and cyclin E1 in a time-dependent manner (P＜0.05) and peaked at 30 min (P＜0.05). Different doses of Forskolin (0-20 μmol•L-1) increased the mRNA expression of cyclin D1 and cyclin E1 (P＜0.05), but lower compared with FSH alone. Rp-cAMP (0-40 μmol•L-1), H-89 (0-30 μmol•L-1) and Verapamil (0-100 μg•mL-1) inhibited FSH induced the mRNA expression of cyclin D1 and cyclin E1 in a dose-dependent manner (P＜0.05), and the combination of Rp-cAMP and Verapamil had more inhibition on FSH activity than individual effect (P＜0.05), but lower than addition of Rp-cAMP and Verapamil. Furthermore, different doses of U0126 (0-10 μmol•L-1) reduced FSH induced the mRNA expression of cyclin D1 and cyclin E1 (P＜0.05), while Rp-cAMP, H-89, Verapamil or U0126 alone had no effect on the mRNA expression of cyclin D1 and cyclin E1 as compared to the control (P＞0.05). 【Conclusion】 FSH regulates the mRNA expression of cyclin D1 and cyclin E1 in a dose- and time-dependent manner, while cAMP-PKA, Ca2+ and extracellular signal-regulated kinase (ERK1/2) is involved in the regulation of FSH on the mRNA expression of cyclin D1 and cyclin E1. Reference | Related Articles | Metrics Cited: Baidu(2)

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Tissue Expression Profile and Bioinformatics Analysis of Abundance miR-101 from the Hypothalamus of Chicken LI Guo-Xi, WANG Le-Le, SUN Gui-Rong, KANG Xiang-Tao Scientia Agricultura Sinica    2013, 46 (6): 1247-1255.   DOI: 10.3864/j.issn.0578-1752.2013.06.019 Abstract （678）      PDF （578KB）（770）       Knowledge map    Save 【Objective】 The objective of this study is to comprehensively understand the chicken hypothalamus abundance miR-101 expression and potential biological function which will lay a foundation for revealing the molecular mechanisms of hypothalamic regulation of the chicken energy balance.【Method】 Stem-loop quantitative RT-PCR was used to detect the miR-101 expression in 13 different tissues of Gu-shi chicken at four developmental stages. The algorithms PicTar was used to predict miR-101 putative target genes. Gene Ontology, pathway and stratified enrichment were analyzed respectively to target genes. 【Result】 MiR-101 expression exhibitted striking temporal and tissue specificities. The miR-101 expression levels in different tissues of embryonic stages were significantly higher than the expression levels in homologous tissues of hatched chicks, the expression levels in the brain tissues were also significantly higher than the other tissues examined. MiR-101 had abundance enrichment in hypothalamus at 14 d embryonic age and in small intestine and crura muscle at 17 d embryonic age. Predicted target genes of miR-101 were significantly enriched in biological regulation, metabolic processes, developmental processes and cellular processes. Bioinformatics analysis of the nervous system development displayed that the regulatory function of miR-101 was mainly involved in brain development, neuron differentiation, regulation of neurogenesis, glial cell differentiation, astrocyte differentiation, and so on. 【Conclusion】 MiR-101 was the high expression miRNA in brain tissue, and may play an important role in regulating biological processes related to brain development. Reference | Related Articles | Metrics

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Preparation and Characterization of Ractopamine-Imprinted Material Using Surface-Molecular Imprinting Method and Its Adsorption Characteristics YANG Ting, WU Yin-Liang, LI Cun, ZHAO Jian, CHEN Guo, LU Yan Scientia Agricultura Sinica    2013, 46 (6): 1256-1262.   DOI: 10.3864/j.issn.0578-1752.2013.06.020 Abstract （704）      PDF （704KB）（985）       Knowledge map    Save 【Objective】 The objective of this experiment is to prepare a molecule-imprinted material on surface of silica gel for ractopamine. 【Method】 Polymethylacrylic acid (PMAA) was grafted onto the surface of silica gel particles via the coupling effect of γ-methacryloxypropyl trimethoxy silane, and the grafting particle PMAA/SiO2 was prepared. Then, the molecular imprinting towards the grafted PMAA was performed with ractopamine as template molecule and ethylene glycol diglycidylether (EGDE) as crosslinking agent by using the surface-molecular imprinting technique established, and the ractopamine molecule- imprinted material MIP-PMAA/SiO2 was prepared. The morphology of MIP-PMAA/SiO2 was observed by SEM，and the chemical structure of MIP-PMAA/SiO2 was characterized by FT-IR. Static method was adopted to study the binding properties and molecule recognition characters of MIP-PMAA/SiO2 for ractopamine.【Result】The MIP-PMAA/SiO2 on the adsorption ability of ractopamine was significantly better than clenbuterol and salbutamol. The adsorption capacity of the MIP-PMAA/SiO2 was 11.08 mg•g-1, the specificity binding capacity was 9.93 mg•g-1, and the imprinted factor of the polymer was 9.63. The adsorption reaction reached equilibrium after 10 min, and the desorption ration of ractopamine reached 92.3% in 5 min.【Conclusion】The experimental results show that MIP-PMAA/SiO2 have specific recognition selectivity, excellent binding affinity and elution property for ractopamine. Reference | Related Articles | Metrics Cited: Baidu(2)

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Upregulation and Expression of Bombyx mori bmo-miR-14 and Prediction of Its Target Genes LIU Yue, YANG Lan-Cui, NIE Zuo-Ming, LU Xuan, LU Zheng-Bing , CHEN Jian, YU Wei, WU Xiang-Fu, ZHANG Yao-Zhou Scientia Agricultura Sinica    2013, 46 (6): 1263-1271.   DOI: 10.3864/j.issn.0578-1752.2013.06.021 Abstract （594）      PDF （645KB）（843）       Knowledge map    Save 【Objective】The objective of this study is to offer a method for upregulating miRNA and predicting its targets in BmN cells, and to provide a methodological reference for studying the function of miRNAs in BmN cells. 【Method】An eukaryotic expression vector pSK-hr3-ie1-EGFP-pri-mir-14 for expressing Bombyx mori bmo-miR-14 was constructed and the miRNA gene was expressed in BmN cells. Additionally, the level of bmo-miR-14 in BmN cells was also up-regulated by transfecting chemically synthesized miRNA mimics. Firstly, the fragment containing pri-mir-14 was amplified by PCR and inserted into the downstream of EGFP of eukaryotic expression vector pSK-hr3-ie1-EGFP. The recombinant plasmid pSK-hr3-ie1-EGFP-pri-mir-14, control plasmid pSK-hr3-ie1-EGFP, bmo-miR-14 mimics and negative control mimics were transfected into BmN cells, respectively. The fluorescent light was observed and used to identify the efficiency of transfection. The miRNA levels in transfected BmN cells were identified by qRT-PCR. The RNAhybrid and miRanda were used to predict the targets of bmo-miR-14, respectively. 【Result】The miR-14 level was improved in BmN cells by transfecting with pSK-hr3-ie1-EGFP-pri-mir-14 and bmo-miR-14 mimics, and the level was improved as 2.1 and 984 times as the control, respectively. Further, the targeted genes of bmo-miR-14 were identified by bioinformatics tools and a total of 153 and 171 potential targeted genes were identified by RNAhybrid and miRanda software, respectively. In the predicted target genes, a total of 49 genes were predicted by RNAhybrid and miRanda together. The 49 targeted genes were assigned to GO terms of molecular function ontology, and binding and catalytic were overrepresented. In the biological process ontology, cellular process and metabolic process overrepresented among the 49 genes.【Conclusion】The approach to upregulate the level of bmo-miR-14 by transfecting with recombinant expression vector or miRNA mimics has laid a good foundation for the further studies on bmo-miR-14 in BmN cells. Reference | Related Articles | Metrics Cited: Baidu(1)

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Cloning and Testicular Expression Level of b-Sycp2 in Cattle, Yak and Cattle-Yak LUO Hua, JIA Chao, QU Xu-Guang, ZHAO Xing-Bo, ZHONG Jin-Cheng, XIE Zhuang, LI Qi-Fa Scientia Agricultura Sinica    2013, 46 (2): 367-375.   DOI: 10.3864/j.issn.0578-1752.2013.02.016 Abstract （817）      PDF （825KB）（676）       Knowledge map    Save 【Objective】The study was aimed to clone the Sycp2 gene sequence of cattle, yak and cattle-yak, investigate the gene structures and its tissue-expression patterns and testicular expression levels in cattle, yak and cattle-yak.【Method】 In silico cloning and clone sequencing were applied to acquire the coding region sequences of Sycp2 gene of cattle, yak and yak-cattle, and molecular characterization were analyzed by bioinformatics software. RT-PCR was applied to analyze the tissue-expression patterns, and real-time PCR was employed to examine the expression levels in cattle, yak and cattle-yak testis.【Result】The full length of the coding region sequences of the Sycp2 gene was 4 365 bp, named b-Sycp2. The b-Sycp2 gene in cattle, yak and cattle-yak encoded 1 454 amino acid residues which included some typical domains such as coiled-coil domain. RT-PCR analysis showed that b-Sycp2 gene was expressed only in the testis. Real-time PCR analysis revealed that the mRNA expression level of b-Sycp2 gene in the testis of cattle and yak was remarkably higher than that of in cattle-yak (P＜0.05). 【Conclusion】The b-Sycp2 gene was cloned successfully, and it expressed only in testis, b-Sycp2 mRNA expression level of cattle and yak represented a dramatic higher than that of cattle-yak. Reference | Related Articles | Metrics Cited: Baidu(1)

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Screening Differentially Expressed Genes of Skin Tissue of Different Flowers Patterns of Hu Sheep SUN Wei, NI Rong, YIN Jin-Feng, DING Jia-Tong, ZHANG You-Fa, CHEN Ling, WU Wen-Zhong, ZHOU Hong Scientia Agricultura Sinica    2013, 46 (2): 376-384.   DOI: 10.3864/j.issn.0578-1752.2013.02.017 Abstract （878）      PDF （571KB）（1001）       Knowledge map    Save 【Objective】The molecular genetic mechanism of China’s unique white lambskin breed different flowers patterns of Hu sheep and hair follicle formation were studied.【Method】The article focused on screening differentially expressed genes of new born Hu-sheep’s skin tissue by Agilent gene chip.【Result】 Differential expression analysis revealed that 1067, 2071 and 3879 differentially expressed genes could be screened from 3 groups of large flowers and small flowers, and compared the 3 groups it could find that there were 137 genes in common. Differentially expressed genes classified by GO (Gene Ontology) analysis mainly involved in cell differentiation, proliferation, apoptosis, growth, immune response, ion transport etc. biological progresses. In addition, the RT-PCR results of 3 differentially expressed genes were consistent with that of gene chip.【Conclusion】Combined with the related literatures, BMP7, MMP2, SNAI1, SFXN1, CDKNIC, MT3, POU1F1 etc. might have important effects on the formation of large flowers and small flowers and could be regarded as candidate genes, and offered the basic data for the further study on the molecule genetic mechanism of hair follicles’ formation in Hu sheep. Reference | Related Articles | Metrics Cited: Baidu(3)

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Transfected FST Gene into Ovine Muscle-Drived Myogenic Cells to Regulate the Expression of MSTN Gene YUE Qun-Hua, YUAN Jian-Long, HAO Fei, JIN Mu-Zi, WANG Jing-Yuan, YANG Wen-Liang, LIU Dong-Jun, CANG Ming Scientia Agricultura Sinica    2013, 46 (2): 385-393.   DOI: 10.3864/j.issn.0578-1752.2013.02.018 Abstract （502）      PDF （702KB）（700）       Knowledge map    Save 【Objective】In this research, skeletal muscle specific expressed follistatin（FST）eukaryotic expression vectors, pCFCDs-red and pSPFCDs-red, were constructed, then respectively transfected into different cell lines. FST and downstream gene expression situation in RNA and protein levels was detected, and the simultaneous start efficiency of skeletal muscle-specific promoter of sub-SP and α-actin was assayed, so that to understand the effect of specific expression of FST on the development of muscle.【Method】Using liposomes, the eukaryotic expression vectors of pCFCDs-red and pSPFCDs-red were transfected respectively into sheep fetal fibroblast cells (SFFCs), skeletal muscle satellite cells (SMSCs) and SMSCs induced muscle tube, and the transgenic cells were analyzed on status of cell growth, cell cycle, cell size and expression of target gene and downstream gene respectively by flow cytometry, Real-Time quantitative PCR and western blotting. 【Result】The growth of transgenic cells basically comply with the trend of non-transgenic cells, and transgenic SMSCs cell had a higher proliferation rates than that genetically modified SFFC. Flow cytometry analysis showed that more than 70% of cells of transgenic cell lines were in G0/G1 phase, maintained a strong ability to divide and had a single peak ,a good aneuploidy and volume increased significantly. Real-time quantitative PCR and western blotting results showed that RNA and protein levels of FST in the transgenic cell lines were raised compared to non-transgenic cell lines. Transgenic SMSCs and myotubes compared to transgenic SFFCs cell had higher expression of FST. Transfected pSPFCDs-red transgenic cells had higher expression level of FST than transfected pCFCDs-red cells.【Conclusion】Skeletal muscle specific promoter SP and α-actin can effectively start FST target gene in the SMSCs and myotubes, and FST have a higher expression level in myotubes. Start efficiency of SP promoter is higher than that of α-actin. In myogenic cells, the increased FST can cause the reduction of MSTN proteins level. FST has a certain role in promoting growth and development of skeletal muscle cells. Key words: FST; synthetic promoter SP ;specific promoter α-actin; skeletal muscle satellite cell; myotubes Reference | Related Articles | Metrics Cited: Baidu(5)

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Comparative Analysis of Proteome and Phosphoproteome of Drone Embryos Between Apis cerana cerana and A. mellifera ligustica ZHOU Tian-E, FANG Yu, FENG Mao, HAN Bin, ZHANG Lan, LI Rong-Li, LI Jian-Ke Scientia Agricultura Sinica    2013, 46 (2): 394-408.   DOI: 10.3864/j.issn.0578-1752.2013.02.019 Abstract （636）      PDF （1740KB）（414）       Knowledge map    Save 【Objective】The objective of this study is to compare the proteome and phosphoproteomic during the three days development of Apis cerana cerana and A. mellifera ligustica, investigate the characteristics of embryonic development and gain an in-depth understanding of the developmental biology of the drone embryos, and to lay a foundation for the future genetic improvement of honeybees.【Method】Two-dimensional gel electrophoresis (2-DE) was employed to compare the proteomic and phosphoproteomic differences between the drone embryogenesis of the two bee species at three time-points of 24, 48 and 72 h, respectively. The level of protein expression, pI and molecular weight were determined. Furthermore, some of the differentially expressed proteins were analyzed by mass spectrometry and bioinformatics.【Result】In total proteome, 332, 362 and 340 protein spots were detected in drone embryos of A. cerana cerana from day 1 to day 3. Of these, 111, 117 and 112 proteins were phosphorylated, respectively. In the drone embryos of A. mellifera ligustica, 302, 331 and 311 proteins were detected, in which 107, 110 and 106 proteins were phosphorylated. Meanwhile, 214, 239 and 220 proteins were commonly expressed during the first three days of drone embryonic development. Thirty differentially expressed proteins were involved in carbohydrate metabolism and energy production, protein folding, development, regulatory proteins and cytoskeletal proteins.【Conclusion】A larger number of proteins and phosphoproteins were expressed in A. cerana cerana than those in A. mellifera ligustica during the drone embryogenesis, indicating A. cerana cerana needs more protein to regulate its drone embryonic development. About 1/3 protein phosphorylated in both honeybee embryos might be associated with signal transduction, cell proliferation and differentiation during drone embryogenesis. Embryos of A. mellifera ligustica expressed higher proportion of housekeeping proteins than in A. cerana cerana suggest the embryogenesis of A. mellifera ligustica has shaped more conservative manner of development due to its longer evolution history. Function analysis indicates that in the long-term evolutionary process has shaped the two honeybee species perusing their own embryonic development ways to fit their biological characteristics. Some of the important biological characteristics, such as A. cerana cerana is good at collecting sporadic nectar and pollen and defending wasps, and A. mellifera ligustica has higher production of honey and royal jelly, have been partially unraveled by this proteome dataset. Related Articles | Metrics

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Construction of Eukaryotic Expression Vector of Bovine fabp3 or fabp4 Gene and Expression of the Gene in Mouse Myoblasts WANG Jie, AO Xu-Dong, LI Wen-Bin, BAI Hai-Dong, YUE Yong-Li, YU Hai-Quan Scientia Agricultura Sinica    2013, 46 (1): 136-145.   DOI: 10.3864/j.issn.0578-1752.2013.01.016 Abstract （639）      PDF （752KB）（600）       Knowledge map    Save 【Objective】 The objective of the study is to analyze the intrinsic expression bovine fabp3 and fabp4 genes, and the dynamic changes of fabp3 and fabp4 gene expression after transfected the eukaryotic expression vector into mice myoblasts. 【Method】 Eukaryotic expression vector pDsHF3 for fabp3 gene, pDsHF4 for fabp4 gene were constructed and transfected into mice myoblasts by using liposome technique. The transfected mice myoblasts were induced differentiation into myotubes by 2% of pregnant mare serum after 72 h transfection. The changes of gene expression of myoblasts and myotubes were analyzed by real-time PCR. 【Result】 The results showed that, bovine fabp3 or fabp4 genes were expressed in transfected myoblasts, respectively. The intrinsic expression of mouse fabp3 or fabp4 gene was affected in myoblasts and in differentiated myotubes.【Conclusion】 Eukaryotic expression vectors pDsHF3 and pDsHF4 can highly express in proliferative myoblasts, and the expression of exogenous bovine fabp3 or fabp4 genes may affect intrinsic expression of mouse fabp3 and fabp4 genes in myoblasts and myotubes. Reference | Related Articles | Metrics Cited: Baidu(1)

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MicroRNAs in Ovaries of Goats (Capra hircus) Identified by Solexa Sequencing ZHANG Xiao-Dong, LING Ying-Hui, ZHANG Yun-Hai, LI Yun-Sheng, LIU Ya, CAO Hong-Guo, ZHANG Zi-Jun, YIN Zong-Jun, DING Jian-Ping, ZHANG Xiao-Rong Scientia Agricultura Sinica    2013, 46 (1): 146-153.   DOI: 10.3864/j.issn.0578-1752.2013.01.017 Abstract （686）      PDF （494KB）（1185）       Knowledge map    Save 【Objective】 Characterization of microRNA expression in hircine ovary tissues was studied, with the purposes of providing basic information for further studies of relationships between specific miRNAs and hircine reproduction such as folliculogenesis and sex hormone secretion. 【Method】 The small RNAs isolated from total RNA of ovary tissues were sequenced by Solexa and then bioinformatics analysis were perfomed. Meanwhile, the expression of selected miRNAs was validated by q-PCR. 【Result】Sequence analysis indicated that 508 ovary miRNAs and 19 corresponding miRNA*s were identified which are conservative in evolution of mammals (sheep, bovine, swine, horse, dog). The expression of 8 selected miRNAs in ovary tissues obtained by q-PCR was in agreement with Solexa sequencing results. 【Conclusion】 The expression profiling of miRNAs, which are abundant and differentially expressed in hircine ovary tissues, was constructed successfully. Reference | Related Articles | Metrics Cited: Baidu(5)

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Interpretation of Odorant Binding Function and Mode of General Odorant Binding Protein ASP2 in Chinese Honeybee (Apis cerana cerana) LI Hong-Liang, ZHANG Lin-Ya, ZHUANG Shu-Lin, NI Cui-Xia, HAN Bao-Yu, SHANG Han-Wu Scientia Agricultura Sinica    2013, 46 (1): 154-161.   DOI: 10.3864/j.issn.0578-1752.2013.01.018 Abstract （664）      PDF （827KB）（985）       Knowledge map    Save 【Objective】 The objective of this study is to research the binding function and mode of different odors with general odorant binding protein AcerASP2 of Chinese honeybees (Apis cerana cerana). 【Method】 With the optimization of induction conditions, the purified recombinant AcerASP2 protein was obtained, then the competitive fluorescence assay was used to determine the binding function of AcerASP2 with odors having different structures, finally the homology modeling and molecular docking were applied to elucidate the binding mode and mechanism. 【Result】After the soluble recombinant AcerASP2 protein purified, in the competitive fluorescence assay between AcerASP2 and 1-NPN, the dissociation constants K1-NPN and the number of binding sites n were 7.38 μmol•L-1 and 1.0321, respectively. In selective 7 kinds of odors, the affinity of 4-allylveratrole seemed to be the strongest, and the IC50 and dissociation constant KD were 7.09 and 3.46 μmol•L-1, respectively. The molecular docking results showed that AcerASP2 had one elongated pocket-like hydrophobic cavity, 4-allylveratrole just existed in the cavity, and two hydrogen bonds were found with Lys74 of AcerASP2. 【Conclusion】Because of the particular spatial structure of binding cavity, AcerASP2 could bind diverse odors with different structures, and the binding interaction could be easily to be promoted by the hydrogen bonds between odors with lysines in AcerASP2. Reference | Related Articles | Metrics Cited: Baidu(14)

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Energy Requirement Parameters of 20-35 kg Dorper and Thin-Tailed Han Sheep Crossbred Male Lambs XU Gui-Shan, DIAO Qi-Yu, JI Shou-Kun, DENG Kai-Dong, JIANG Cheng-Gang, TU Yan, LIU Jie, ZHAO Yi-Guang, MA Tao, LOU Can Scientia Agricultura Sinica    2012, 45 (24): 5082-5090.   DOI: 10.3864/j.issn.0578-1752.2012.24.014 Abstract （559）      PDF （570KB）（902）       Knowledge map    Save 【Objective】 The research aimed to define the energy requirement for maintenance and growth of metabolic energy and net energy for 20-35 kg Dorper and Thin-Tailed Han crossbred F1 ram lambs, to provide a theoretical basis of scientific feeding mutton ram lambs. 【Method】 The trial was conducted according to a completely random design and lasted for 66 days. Fifty Dorper and Thin-Tailed Han ram lambs crossbred F1 lambs were selected, thirty five lambs were used for slaughter trail and 15 lambs were used for digestibility trial. Seven lambs were randomly chosen and slaughtered at (20.26±1.29) kg BW for measuring the initial body composition. The remaining 28 ram lambs were offered a pelleted mixture diet for ad libitum intake, or 70 or 40% of the ad libitum intake. Seven of the ram lambs were randomly chosen and slaughtered when the 14 ram lambs fed ad libitum reached (28.54±2.29) kg. The remaining 21 ram lambs were randomly divided into 3 intake levels and fed a pelleted mixture diet, and the lambs were all slaughtered when the lambs in ad libitum treatment group reached at 35 kg of BW. In a digestibility trial, fifteen lambs (32.38±2.23) kg were randomly divided into three groups (five lambs in each group), the feed and the intake level were the same as slaughter experiment. Total collection of feces and urine was conducted. Methane production, carbon dioxide output and oxygen consumption were measured by open-circuit respirometry. 【Result】 The results showed that the maintenance requirements for NE and ME were 250.61 and 374.21 kJ•kg-1 metabolic shrunk BW (SBW0.75), respectively, with a partial efficiency of energy utilization for maintenance being 0.67. Net energy and metabolic energy requirements for growth ranged from 1.12 to 5.31 MJ•d-1 and 2.63 to 12.03 MJ•d-1, respectively, for the lambs gaining 100 to 350 g•d-1 from 20 to 35 kg body weight. The partial efficiency of ME for growth was 0.419. 【Conclusion】 Maintenance requirement for net energy and metabolic energy, net requirements and metabolic energy for gain of 20-35 kg Dorper and Thin-Tailed Han ram lambs crossbred F1 lambs were determined. These parameters were slightly lower than those reported by NRC and AFRC. Reference | Related Articles | Metrics Cited: Baidu(9)

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Cloning, Sequence Analysis and Over-Expression of SCD Gene of Dairy Goat SHI Heng-Bo, LUO Jun, ZHU Yue, WANG Zi-Qian, ZHAO Wang-Sheng Scientia Agricultura Sinica    2012, 45 (24): 5091-5101.   DOI: 10.3864/j.issn.0578-1752.2012.24.015 Abstract （564）      PDF （744KB）（1414）       Knowledge map    Save 【Objective】To study the function of stearoyl-CoA desaturase (SCD) gene in mammary gland fatty acid metabolism emphasizing on how it regulates genes involved in unsaturated fatty acid synthesis as well as its impact on fatty acid composition.【Method】The sequence of SCD in dairy goat was cloned using RT-PCR for bioinformatics and tissue mRNA expression analysis. A recombinant adenovirus vector was constructed to over-express the gene in cultured goat mammary epithelial cells. Changes of gene expression, protein yield and fatty acid contents after over-expression were detected with real-time fluorescence quantitative PCR, western blotting and GC-MS. 【Result】The CDS of goat SCD gene is 1080 bp in length, coding 359 amino acid residues (GenBank accession: GU947654). The SCD gene was expressed higher in mammary gland, lung and subcutaneous fat, lower in heart, rumen and ovary. Recombinant adenovirus vector containing goat SCD open reading frame was constructed, packaged and amplified in HEK293. The gene expressions of FASN, A-FABP, LEPR and LXRα were down regulated after over-expression of SCD, while expressions of PPARγ and LPL were up regulated significantly. Synthesis of palmitic acid and stearic acid in the cells decreased, while unsaturated fatty acids increased.【Conclusion】 SCD gene plays an important role in goat mammary fatty acid metabolism, and a major role in unsaturated fatty acid synthesis by interaction with other genes including FASN, H-FABP, LEPR, LXRα, PPARγ and LPL. Reference | Related Articles | Metrics Cited: Baidu(8)

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Investigation on Contamination Situation of Aflatoxin in Detected Feeds and Feedstuffs in Beijing Area FAN Yu, LI Xiao-Ying, ZHAO Li-Hong, JIA Ya-Xiong, JI Cheng, MA Qiu-Gang, CHEN Yu, WANG Liang Scientia Agricultura Sinica    2012, 45 (24): 5102-5109.   DOI: 10.3864/j.issn.0578-1752.2012.24.016 Abstract （799）      PDF （551KB）（891）       Knowledge map    Save 【Objective】This study was conducted to analyze and evaluate the contamination situation of aflatoxin in feeds and feedstuffs in Beijing area. 【Method】 A total of 187 feed samples from 21 farms in Beijing were sampled to detect and quantify the content of aflatoxin (B1, B2, G1 and G2). An efficient HPLC with post-column photochemical derivatization method was developed, including immunoaffinity step for aflatoxin extraction from feeds and feedstuffs followed by liquid chromatography (LC) for quantification. 【Result】 Results showed that the detection rate of AFB1 in corn, bran, soybean meal, DDGS, swine and poultry complete feeds was 50.0%, 46.2%, 33.3%, 94.1%, 67.1% and 94.3%, the over standard rate was 6.2%, 0.0%, 0.0%, 5.9%, 6.6% and 0.0%, and the average content was 5.98, 0.25, 1.00, 9.83, 2.89 and 1.06 µg•kg-1, respectively. 【Conclusion】 These results indicated that the average content of aflatoxin in complete feeds, corn and DDGS were the highest. Aflatoxin contamination was widely found in both feeds and feedstuffs. Reference | Related Articles | Metrics Cited: Baidu(5)

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Cloning and mRNA Expression of Smad4 Gene in Ovaries of Erhualian Pig ZHAO Yong-Xiang, LIU Ji-Ying, PAN Zeng-Xiang, ZHANG Jiu-Feng, YAO Yong, ZHOU Ji-Long, XIE Zhuang, XU Yin-Xue, LIU Hong-Lin, LI Qi-Fa Scientia Agricultura Sinica    2012, 45 (23): 4883-4890.   DOI: 10.3864/j.issn.0578-1752.2012.23.015 Abstract （684）      PDF （895KB）（971）       Knowledge map    Save 【Objective】To investigate The sequence features and tissue distribution patterns of Smad4 gene in Erhualian pigs were investigated, and the Smad4 mRNA expression differences in ovarian tissue between Erhualian and commercial cross pigs were analyzed. 【Method】The cDNA sequence of Smad4 gene was obtained by cloning and sequencing techniques in Erhualian pigs, the gene sequence characteristics, physical and chemical properties and the 3D structure of Smad4 protein were analyzed by using bioinformatics methods. The tissue distribution patterns of Smad4 gene were detected by RT-PCR, and then the mRNA expression levels were detected in ovarian tissue of Erhualian and commercial cross pigs by real-time PCR.【Result】The full length of Smad4 gene coding sequence was 1 659 bp in Erhualian pigs, encoding for a protein with 552 amino acid residues. The coding amino acid sequence was highly conservative with other mammals (more than 98% identity). Same as other mammals, Smad4 protein in Erhualian pigs also had three typical structural domains (MH1, SAD and MH2). RT-PCR assays revealed that Smad4 gene of Erhualian pigs widely expressed in all tissues. In addition, Smad4 mRNA levels in Erhualian pig ovary was significantly higher than that in commercial pigs (P＜0.01). 【Conclusion】 The Smad4 gene in Erhualian pigs may be own the same function as other mammals and exert probably associated with the pig prolificacy. Reference | Related Articles | Metrics Cited: Baidu(1)

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Study on Variation Characteristics of Raw Milk Composition and Curve Models of Chinese Holstein in the City of Tianjin XIONG Ben-Hai, MA Yi, PANG Zhi-Hong, YANG Lu, YI Miao, YANG Qin Scientia Agricultura Sinica    2012, 45 (23): 4891-4897.   DOI: 10.3864/j.issn.0578-1752.2012.23.016 Abstract （775）      PDF （536KB）（793）       Knowledge map    Save 【Objective】 To fulfill the need of seasonal management modification of lactating cows. 【Method】 the effect of natural month, parity, and their interaction on milk components were analyzed in the present study. Data were collected from a DHI database of lactating Chinese Holstein cows in the north part of china. Original data were screened and group according to parity (1-4). A total of 6114 milk protein content records and 5871 milk fat content records were analyzed by GLM procedure of SAS. 【Result】The Duncan multiple comparison of natural months, regardless of parity (only parity 1 to 4), demonstrated that the milk compositions of different months showed significant difference (P＜0.05), although the data between some different months showed no significant difference. The milk protein percentage in September reached the highest (3.187%), and in July the lowest (3.016%). Milk fat percentage in February was the highest (4.137%), and in July the lowest (3.845%).The same multiple comparison of different parity, also regardless of different months (1-12 months), demonstrated that the milk composition data of different parities showed significant difference (P＜0.05), although the data between some parities showed no significant difference. The milk protein percentage reached the highest in parity 2 (3.114%), and the lowest in parity 4 (3.066%). The milk fat percentage reached highest in parity 2 (3.983%) and parity 3 (3.973%), respectively; and the lowest in parity 4 (3.923%). In addition, the relation equation between the milk protein percentage (MPP, %) or the milk fat percentage (MFP, %) of different parities and the natural months in mixed cow herd was built using Model Wood, i.e. MPP=3.094x-0.0464×e0.0117x, MFP=4.2116x-0.0344×e0.0276x, x as month. 【Conclusion】The natural months, milking parities and their interaction had significant affects on milk composition including milk protein percentage and milk fat percentage, and milk composition had wood pattern changing relationship with natural months respectively. Reference | Related Articles | Metrics Cited: Baidu(4)

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Cloning, Expression and Microbial-Induction Analysis of Bombyx mori Serine Protease Inhibitor BmSPI37 LI You-Shan, ZHAO Ping, DONG Zhao-Ming, ZOU Yong, XIA Qing-You, XIANG Zhong-Huai Scientia Agricultura Sinica    2012, 45 (23): 4898-4908.   DOI: 10.3864/j.issn.0578-1752.2012.23.017 Abstract （655）      PDF （976KB）（827）       Knowledge map    Save 【Objective】 The objective of this study is to identify a new protease inhibitor of silkworm (Bombyx mori) and to investigate its function in guarding against invasion of pathogenic microorganisms. 【Method】 T-A cloning, multiple sequence alignment, construction of phylogenetic tree, prokaryotic expression, spatio-temporal expression profile analysis and microbial-induced experiments were performed with BmSPI37. 【Result】 A new serine proteinase inhibitor, named BmSPI37, was cloned and expressed. Spatio-temporal expression profile analysis showed that BmSPI37 expressed highly at the late stage of 5th instar larvae, and expressed highly in the middle silk gland. The expression of BmSPI37 was significantly higher in female than in male during metamorphosis period from pupa to moth. Analysis of the expression of BmSPI37 after infection with Escherichia coli, Bacillus bombysepticus or Beauveria bassiana showed that BmSPI37 was strongly up-regulated.【Conclusion】It is speculated that BmSPI37 is related to defense against pathogenic microorganisms. Reference | Related Articles | Metrics

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Effect of Royal Jelly on the DNA Methylation of dynactin p62 in Female Honeybee Larvae SHI Yuan-Yuan, TIAN Liu-Qing, ZHANG Fei, LIU Jun-Feng, YAN Wei-Yu, ZENG Zhi-Jiang Scientia Agricultura Sinica    2012, 45 (23): 4909-4915.   DOI: 10.3864/j.issn.0578-1752.2012.23.018 Abstract （866）      PDF （535KB）（1020）       Knowledge map    Save 【Objective】 The objective of this study is to analyze the influence of royal jelly on DNA methylation of dynactin p62 in female honeybee larvae. 【Method】 One-day-old larvae of Apis mellifera ligustica and A. cerana cerana were used as materials, they were fed with royal jelly either from A. mellifera (RJM) or from A. cerana (RJC), and genome DNA was extracted from 3-and 6-day-old larvae, then DNA methylation of dynactin p62 was detected by Sequenom MassARRAY. 【Result】 The whole DNA methylation of dynactin p62 in female honeybee larvae decreased significantly after feeding with heterospecific royal jelly. The whole DNA methylation of dynactin p62 in 6-day-old larvae was significantly lower than 3-day-old larvae after feeding with homologous royal jelly. The methylated sites of dynactin p62 in larvae were different after feeding with royal jelly (RJM and RJC). 【Conclusion】 Royal jelly (RJM and RJC) had different effects on the whole DNA methylated and methylated sites of dynactin p62 in female honeybee larvae. These results indicate that different types of royal jelly have different biological effects. Reference | Related Articles | Metrics Cited: Baidu(5)

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Prokaryotic Expression of Bombyx mori Insulin-Related Peptide-Binding Protein 2 (BmIBP2) and Its Tissue-Specificity in Silkworm GAO Kun, DENG Xiang-Yuan, ZHU Feng, WANG Sheng-Peng, QIAN He-Ying, GUO Xi-Jie Scientia Agricultura Sinica    2012, 45 (23): 4916-4923.   DOI: 10.3864/j.issn.0578-1752.2012.23.019 Abstract （612）      PDF （687KB）（558）       Knowledge map    Save 【Objective】 The objective of this study is to express insulin-related peptide-binding protein 2 from silkworm (BmIBP2) which belongs to the immunoglobulin (Ig) superfamily in vitro and prepare polyclonal antibody against recombinant BmIBP2 for the further research about its biological function．【Method】 The gene fragment of the mature BmIBP2 peptide was amplified by PCR method with the cDNA synthesized from silkworm midgut. In order to express the protein of mature BmIBP2 peptide, the DNA segment was inserted into the expression plasmid pET-28a(+) to construct a recombinant expression plasmid. Polyclonal antibody was prepared to study the tissue-specificity expression of BmIBP2 protein in silkworm midgut. 【Result】 The SDS-PAGE result showed that the cloned recombinant BmIBP2 (rBmIBP2) was expressed in the form of inclusion bodies in E. coli BL21 with a molecular weight of 30 kD, and Western blot using the antibody against His-tag as the primary antibody indicated that rBmIBP2 had satisfied immunobiological activity. Pure protein was obtained by Ni2+ NTA resin column to prepare the anti-rBmIBP2 polyclonal antibody by immunizing New Zealand white rabbit. Western blot analysis showed that BmIBP2 protein could be bound with the polyclonal antibody in silkworm midgut with a molecular weight of 28 kD, which is consistent with the predicted molecular of BmIBP2 mature peptide．However, no specific bands were found in other tissues including fat body, hemocytes, silk gland, testicle and ovary. 【Conclusion】 Recombinant BmIBP2 protein was successfully expressed in E. coli BL21, and its polyelonal antibody was prepared in this study. Moreover, Western blot showed that BmIBP2 was correctly expressed in vitro and had a tissue-specificity expression in silkworm midgut. Reference | Related Articles | Metrics Cited: Baidu(1)

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Effects of Degradable Protein and Non-Fiber Carbohydrates on in Vitro Ruminal Fermentation, Microbial Synthesis, and Populations of Ruminal Cellulolytic Bacteria ZHAO Xiang-Hui, LIU Chan-Juan, LIU Ye, LI Chao-Yun, YAO Jun-Hu Scientia Agricultura Sinica    2012, 45 (22): 4668-4677.   DOI: 10.3864/j.issn.0578-1752.2012.22.013 Abstract （932）      PDF （699KB）（886）       Knowledge map    Save 【Objective】 The objective of this study was to investigate the effects of levels of rumen degradable protein (RDP) and sources of non-fibre carbohydrates (NFC) on ruminal fermentation, microbial synthesis, and populations of ruminal cellulolytic bacteria using the rumen simulation technique (Rusitec). 【Method】 The experiment was a 2×4 factorial arrangement with four NFC types (corn starch, sucrose, pectin, inulin) each combined with two levels of RDP obtained by supplementing with 0 g•d-1 (low RDP) or 1.56 g•d-1 (high RDP) sodium caseinate. 【Result】 Apparent disappearance of dry matter and organic matter was greater (P＜0.01) for the main effect means of sucrose and pectin than for other treatments. A NFC×RDP interaction (P＜0.01) was observed for apparent neutral detergent fibre disappearance, which tended to be lower for sucrose (P=0.10) and pectin (P=0.09) than for starch treatment under low RDP conditions, but did not differ among treatments under high RDP conditions. The 16S rDNA copies of Ruminococcus albus were greater for the main effect means of pectin than for starch treatment. There were NFC×RDP interactions for 16S rDNA copy numbers of R. flavefaciens in both liquid and solid fractions and Fibrobacter succinogenes in liquid fraction. Compared with starch treatment, R. flavefaciens in solid fraction tended to be lower (P=0.06) for sucrose treatment under low RDP conditions and F. succinogenes in liquid fraction was lower (P＜0.01) for inulin treatment under high RDP conditions. Increasing dietary RDP increased total volatile fatty acids production (P＜0.01) and total microbial nitrogen (MN) flow (P＜0.01) in all treatments. The molar proportion of acetate and the ratio between acetate and propionate were both the greatest (P＜0.05 and P＜0.01, respectively) for the main effect means of pectin among treatments. Butyrate molar proportion was greater (P＜0.01) for sucrose and inulin treatments than for other treatments regardless of RDP level. Total MN flow did not differ among treatments under low RDP conditions, but sucrose (P＜0.01) and pectin (P=0.10) produced greater MN than starch with increased RDP. The efficiency of available N was lower for the main effect means of starch than for sucrose (P=0.04) and pectin (P=0.05) treatments.【Conclusion】 Dietary RDP level, NFC type, and their interaction affected ruminal fermentation, microbial synthesis, and cellulolytic bacteria populations, and under sufficient ruminal available N sucrose and pectin had greater advantage in microbial N synthesis than starch. Reference | Related Articles | Metrics

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Developmental Changes of Gene Expression of GHR and IGF-Ⅰ Genes and Their Association Analysis with Meat Quality Traits in Hu Sheep SUN Wei, LI Da, MA Yue-Hui, GUAN Wei-Jun, CHU Ming-Xing, DING Jia-Tong, LI Bi-Chun, ZHANG You-Fa, CHEN Ling, WU Wen-Zhong, ZHOU Hong Scientia Agricultura Sinica    2012, 45 (22): 4678-4687.   DOI: 0.3864/j.issn.0578-1752.2012.22.014 Abstract （547）      PDF （625KB）（853）       Knowledge map    Save 【Objective】The objective of this study was to discuss the developmental changes of gene expression of growth hormone receptor (GHR) and insulin-like growth factor-Ⅰ(IGF-Ⅰ) and their association analysis with meat quality traits , and the relationship of the expression level between GHR gene and IGF-Ⅰgene in Hu sheep. 【Method】The developmental changes of gene expression of GHR gene and IGF-Ⅰgene on the early growth of Hu sheep were detected by real-time fluorescent quantitative RT-PCR. 【Result】The results were as followes: With increasing age after birth, the GHR expression trend of ewes was up-down-up-down-up, and 6-month reached the top. The GHR expression trend of rams was that 2-day-old to 2-month-old was increasing, 2-month-old to 4-month-old was decreasing, 4-month-old to 6-month-old was increasing and reached the top. With the increasing age after birth, the IGF-Ⅰgene expression trend of ewes was increasing from 2-day-old to 6-month-old which had the top expression. The IGF-Ⅰgene expression trend of rams was that at 2-day-old firstly increased to a peak at one-month-old, 1-month-old to 3-month-old had a decreasing process and reached the same level which was close to the newborn, and then increased with the increasing age, and 6-month-old reached the top. Most growth phases had a significant difference or extreme significant difference in GHR and IGF-Ⅰand the expressions of GHR and IGF-Ⅰbetween most rams and ewes that at the same growth phase had a significant difference or extreme significant difference. The results showed that the expressions of GHR and IGF-Ⅰhad a significant positive correlation, (0.01＜P＜0.05) the expressions of GHR and IGF-Ⅰgenes had a extreme significant positive correlation (P＜0.01) with diameter of muscle fibers and a extreme significant negative correlation (P＜0.01) with density of muscle fibers, and the expressions of GHR had a extreme significant positive correlation (P＜0.01) with shear force of muscle fibers, the expression of IGF-Ⅰhad a significant positive correlation with shear force of muscle fibers (0.01＜P＜0.05). 【Conclusion】The results showed that different stages and different genders had important influences on the expressions of GHR and IGF-Ⅰgenes in sheep skeletal muscle. After birth, the expressions of the two genes did not always increase or decrease, the turning points of different genes expression were not at the same time. The expression of GHR and IGF-Ⅰhad a significant positive correlation in early muscle traits of Hu sheep. GHR and IGF-Ⅰgene can be regarded as the candidate gene for the trait of meat quality in Hu Sheep. Reference | Related Articles | Metrics Cited: Baidu(7)

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Construction and Identification of Normalized Subtractive cDNA Library of Plumage Color-Related Genes in Mule Duck ZHENG Nen-Zhu, CHEN Xiao-Yan, LU Li-Zhi, ZHU Zhi-Ming, MIAO Zhong-Wei, XIN Qing-Wu, CHEN Hui, XIAO Tian-Fang Scientia Agricultura Sinica    2012, 45 (22): 4688-4696.   DOI: 10.3864/j.issn.0578-1752.2012.22.015 Abstract （573）      PDF （534KB）（550）       Knowledge map    Save 【Objective】 The aim of this study was to isolate plumage color related-genes in mule duck and explore its molecular mechanism. 【Method】 A normalized subtractive library of plumage color related-genes of mule duck was constructed. White plumage skin and black plumage skin were adopted respectively as the tester and driver, and a duplex-specific nuclease (DSN) normalized subtractive hybridization method was used to construct the library, and real-time PCR was employed to identify the quality of the library.【Result】Results showed that 64 genes with the average size of 1 031 bp were obtained by selecting and sequencing 144 positive clones randomly. Nucleotide BLAST homological analysis incidated that 21 genes had similarities to known genes which enjoy 92.8% homology, and 43 genes were not matched which presumably may be new genes related to plumage color. Geneontology displayed that these known genes were involved in many biological processes such as signal transduction, cell structure, material transport, apoptosis, cell and organism defense, transcription and expression regulation，and had different correlations with formation and transshipment of pigment. 【Conclusion】 It was determined that the library had good quality and could enrich the genes of the white plumage via real-time PCR. Reference | Related Articles | Metrics Cited: Baidu(1)

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Pathogenic Mechanism of Bombus patagiatus Infected by Nosema ceranae QIN Hao-Ran, HE Shao-Yu, WU Jie, LI Ji-Lian Scientia Agricultura Sinica    2012, 45 (22): 4697-4704.   DOI: 10.3864/j.issn.0578-1752.2012.22.016 Abstract （781）      PDF （787KB）（996）       Knowledge map    Save 【Objective】The objective of this study is to define the infectivity and pathogenic mechanism of Nosema ceranae to Bombus patagiatus. 【Method】 Traditional biology and ultrastructure under electronic microscope methods were used, and combined with the quantitative real-time PCR (qPCR) to explore the pathogenesis of N. ceranae to B. patagiatus. 【Result】 In the initial infection, the infected bees did not exhibit obvious external disease signs except decreased feeding and motor retardation. In the late period, the infected bees exhibited dispirited, weak and unable to fly. Through observation with light microscope, few N. ceranae but a lot of bacteria were found in the midgut of bees. N. ceranae mainly infected midgut of epithelial cells, the nuclear enlarged and out of shape, mitochondria become smaller even disintegrating and endoplasmic reticulum become disorders. However, the spores only infected the cytoplasm of the host instead of invading the nuclear, which lead to the disintegration of the mitochondria and cytoclasis. Quantitative analysis on RT-PCR showed that N. ceranae reached the highest level in the midgut and fat body 3-4 d after being infected and other organizations were barely detected.【Conclusion】N. ceranae can cross-species infect B. patagiatus and the pathological process starts with the pathological changes of intestinal cells, which lead to the cytoclasis and apoptosis. Reference | Related Articles | Metrics Cited: Baidu(2)

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Study on Optimization of Non-Starch Polysaccharide Enzymes of Broiler Diets in Vitro HE Ke-Lin, SA Ren-Na, GAO Jie, LI Dong-Wei, ZHUANG Xiao-Feng, ZHANG Hong-Fu Scientia Agricultura Sinica    2012, 45 (21): 4457-4464.   DOI: 10.3864/j.issn.0578-1752.2012.21.014 Abstract （849）      PDF （448KB）（577）       Knowledge map    Save 【Objective】The study was conducted to discuss the effects of non-starch polysaccharides (NSP) enzymes on the in vitro digestive energy (IVDE) of broiler diets of different contents of NSP.【Method】The first step was to investigate the dose-response of the NSP enzymes on the IVDE in diets using one-way randomized experimental design. Five NSP enzymes including xylanase, β-glucanase, cellulose, pectinase, and β-mannanase were added into corn-soybean diet and wheat-soybean diet, from 0 to 900 µg•g-1. The second step was to screen the optimum enzyme combinations using quadratic regress-orthogonal rotary design. The method of testing IVDE was a two-stage enzymic hydrolysis with pepsin-pancreatin in vitro.【Result】The IVDE of broiler diet regulating by NSP enzyme appeared S state diagram. The xylanase had more promotion effects than other NSP enzymes. It increased IVDE by 0.24 MJ•kg-1 and 0.40 MJ•kg-1 in corn-soybean diet and wheat-soybean diet, respectively. In corn-soybean diet, the optimum enzyme combinations were xylanase 34 836.4 U•kg-1, β-glucanase 6 762.0 U•kg-1, cellulose 1 159.2 U•kg-1, pectase 872.6 U•kg-1, and β-mannanase 24 535.9 U•kg-1. In wheat-soybean diet, the optimum enzyme combinations were xylanase 65 405.5 U•kg-1, β-glucanase 10 131.5 U•kg-1, cellulose 980.0 U•kg-1, pectase 501.6 U•kg-1, and β-mannanase 5 141.4 U•kg-1. 【Conclusion】The findings showed that using five kinds of NSP enzymes in corn-soybean and wheat-soybean diets of broiler could improve the in vitro digestive energy. Reference | Related Articles | Metrics Cited: Baidu(3)

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Cloning, Expression and Bioinformatics Analysis of Porcine CatSperB and CatSperG Genes SONG Cheng-Yi, ZHOU Jia-Qing, FENG Xiao-Jun, XIE Yu-Xiu, LI Qing-Ping, WU Han, GAO Bo, WANG Xiao-Yan Scientia Agricultura Sinica    2012, 45 (21): 4465-4474.   DOI: 10.3864/j.issn.0578-1752.2012.21.015 Abstract （676）      PDF （624KB）（700）       Knowledge map    Save 【Objective】The aim of the current study is to confirm the existence of porcine CatSperB and CatSperG genes, and investigate the protein structures, evolutionary relationship and the spatial-temporal expression profiles of CatSperB and CatSperG. 【Method】The in silico and molecular cloning was used to identify the full length cDNAs of porcine CatSperB and CatSperG, and the spatial-temporal expression profile was investigated by qualitative and fluorescence quantitative RT-PCR. 【Result】The in silico transcripts of 3 508 bp CatSperB and 3 715 bp CatSperG were identified, and they contain 3 330 bp and 3 483 bp ORFs of CatSperB and CatSperG, respectively, and the sequences were confirmed by TA cloning. The sequence similarity of coding sequences (CDS) of porcine CatSperB and CatSperG with human, cattle, horse, and dog, and other animals was above 80%. CatSperB is a 125.79 kD and stable protein, while CatSperG is a 133.40 kD and unstable protein. Both CatSperB and CatSperG contain seven conservative trans-membrane domains, and a coiled-coil motif was also identified in the C terminal of CatSperG, but this motif was not found in CatSperB. The porcine CatSperB and CatSperG displayed higher degree of homology with the orthologs of cattle, dog and horse, and lower homology with those of human and mouse. The RT-PCR analysis showed that CatSperB and CatSperG were detected mainly in testis, but CatSperB also expressed in other tissues. The mRNA expression of CatSperB and CatSperG was significantly improved at the around stage of spermatogenesis (Day 60), puberty (Day 90) and sex maturity (Day 150) (P＜0.05). 【Conclusion】The cDNA clones of CatSperB and CatSperG, and a series of bioinformatics parameters of these proteins were obtained. Seven conservative trans-membrane domains in CatSperB and CatSperG and the homology among species were revealed. Furthermore, results of this study had proved that CatSperB and CatSperG expressed mainly in tests, and the change of mRNA expression was paralleled with sexual development of boar. Reference | Related Articles | Metrics Cited: Baidu(1)

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