猪CBR1基因不同拷贝形式克隆及其多态性分析

doi:10.3864/j.issn.0578-1752.2018.08.018

Abstract

Abstract: 【Objective】 In order to obtain and analysis the different copy forms of CBR1 gene coding region, and analysis the correlation between polymorphism and reproductive performance of different copy forms of CBR1 gene in pigs.【Method】 Based on the cDNA of the Wuzhishan pig’s testis , the different copy sequences of CBR1 gene were obtained by cloning sequencing technology. The amino acid sequences encoded by different copy forms of CBR1 gene were analyzed by DNASTAR software. Homology between different copy forms of porcine CBR1 gene, and that compared with other mammals were analyzed using DNAMAN software. The physical and chemical properties of different proteins of porcine CBR1 gene were analyzed by Protparam online software on ExPaSy. 136 Yorkshire and 47 Landrace pig ears samples were selected for DNA extraction and PCR-sequencing to explore the relationship between polymorphism of CBR1 gene and porcine reproductive performance. 【Result】 All the three different copies of CBR1 gene encoded 3 exons, and the sequence length of the coding region were 870bp, 870bp and 846bp respectively. Their 12-18th amino acid residues were GlyXXXGlyXGly, which were in accordance with the CBRs family structural characteristics. The results of homology comparison showed that the CDS sequence homology among different copies of CBR1 gene was more than 87%, and more than 82% among different mammals, indicated that the CBR1 gene of mammal was conserved in evolution process. The physical and chemical properties of 3 different copies of CBR1 were similar, all of them were unstable and belong to hydrophilic proteins, which indicated that they may play a similar role in vivo. Polymorphism analysis showed that 5 SNPs were found in copy V1, 4 SNPs were found in copy V2, and 2 SNPs were found in copy V3. In the Yorkshire pigs, the A/T mutation at the promoter upstream region 153 bp and the C/T mutation at exon3 379 bp in V1; AC insertion at the promoter upstream region 10 bp, C/T mutation at exon 1 63 bp and G/T mutation at intron1 76 bp in V2, were significantly correlated with the multiparous alive litter size, while the SNPs in Landrace pigs were not associated with reproductive performance. 【Conclusion】 Coding region sequence of 3 different copies of CBR1 gene were obtained; 5 SNPs were found significantly associated with multiparous alive litter size , suggested that CBR1 gene may be used as a valuable candidate gene for improving breeding performance of Yorkshire pigs.

Key words: swine, CBR1 gene, re-sequencing, de-novo assembly

QI ChuanXiang, XU KUI, MU YuLian, YANG ShuLin, LI Kui, WU TianWen. The Cloning of Porcine CBR1 Gene’s Different Copy Forms and Analysis of Its Polymorphism[J].Scientia Agricultura Sinica, 2018, 51(8): 1607-1616.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2018.08.018

https://www.chinaagrisci.com/EN/Y2018/V51/I8/1607

References

[1] 吴井生, 王金玉. FSHR基因第10外显子多态性及其与小梅山猪产仔数的相关性. 中国农业科学, 2012, 45(13): 2728-2736.

WU J S, WANG J Y. Polymorphism of exon10 of FSHR gene and its relationship with litter size in Xiaomeishan pigs. Scientia Agricultura Sinica, 2012, 45(13): 2728-2736. (in Chinese)

[2] 马喜山, 王爱国, 刘桂芬, 傅金恋. 猪HB-EGF 基因多态性与产仔数的关联性分析. 中国农业科学, 2009, 42(1): 274-282.

MA X S, WANG A G, LIU G F, FU J L. Analysis of the association between polymorphisms of HB-EGF gene and litter size in pigs. Scientia Agricultura Sinica, 2009, 42(1): 274-282. (in Chinese)

[3] ROTHSCHILD M, JACOBSON C, VASKE D, TUGGLE C, WANG L, SHORT T, ECKARDT G, SASAKI S, VINCENT A, MCLAREN D, SOUTHWOOD O, STEEN H V D, MILEHAM A, PLASTOW G. The estrogen receptor locus is associated with a major gene influencing litter size in pigs. Proceedings of the National Academy of Sciences of the United States of America, 1996, 93(1): 201-205.

[4] 黄贺, 狄生伟, 田亚光, 王希彪, 李和平, 张贵学. 民猪PRLR基因PCR-SSCP多态性与产仔数关联分析. 中国农业科学, 2011, 44(11): 2341-2346.

HUANG H, DI S W, TIAN Y G, WANG X B, LI H P, ZHANG G X. The association analysis between PRLR gene PCR-SSCP polymorphism and litter size in Min pigs. Scientia Agricultura Sinica, 2011, 44(11): 2341-2346. (in Chinese)

[5] 方宇瑜, 马翔, 吴艳, 李强, 王文祥, 王礼伟, 任二都, 李平华, 黄瑞华. 苏淮猪群体ESR1和FSHβ基因的多态性及其与繁殖性状的关联性研究. 畜牧与兽医, 2017, 49(2): 1-5.

FANG Y Y, MA X, WU Y, LI Q, WANG W X, WANG L W, REN E D, LI P H, HUANG R H. Polymorphism analysis of genes ESR1 and FSHβ in Suhuai pigs and correlation analysis with reproductive traits. Animal Husbandry ＆ Veterinary Medicine, 2017, 49(2): 1-5. (in Chinese)

[6] DAI L S, ZHAO Z H, ZHAO R F, XIAO S Q, JIANG H, YUE X P, LI X C, GAO Y, LIU J F, ZHANG J B. Effects of novel single nucleotide polymorphisms of the FSH beta-subunit gene on semen quality and fertility in bulls. Animal Reproduction Science, 2009, 144(3): 14-22.

[7] HAO Y, MURPHY C N, SPATE L, WAX D, ZHONG Z, SAMUEL M, MATHIALAGAN N, SCHATTEN H, PRATHER R S. Osteopontin improves in vitro development of porcine embryos and decreases apoptosis. Molecular Reproduction & Development, 2008, 75 (2): 291-298.

[8] LALIOTIS G P, MARANTIDIS A, AVDI M. Association of BF, RBP4, and ESR2 genotypes with litter size in an autochthonous pig population. Animal Biotechnology, 2017, 28(2): 138-143.

[9] KALABUS J L, SANBORN C C, JAMIL R G, CHENG Q, BLANCO J G. Expression of the anthracycline-metabolizing enzyme carbonyl reductase 1 in hearts from donors with Down syndrome. Drug Metabolism and Disposition, 2010, 38(12): 2096-2099.

[10] RASHID M A, LEE S, TAK E, LEE J, CHOI T G, LEE J W, KIM J B, YOUN J H, KANG I, HA J, KIM S S. Carbonyl reductase 1 protects pancreatic β-cells against oxidative stress-induced apoptosis in glucotoxicity and glucolipotoxicity. Free Radical Biology and Medicine, 2010, 49(10): 1522-1533.

[11] BOTELLA J A, ULSCHMID J K, GRUENEWALD C, MOEHLE C, KRETZSCHMAR D, BECKER K, SCHNEUWLY S. The drosophila carbonyl reductase sniffer prevents oxidative stress-induced neurodegeneration. Current Biology, 2004, 14(9): 782-786.

[12] KALABUS J L, CHENG Q, JAMIL R G, SCHUETZ E G, BLANCO J G. Induction of carbonyl reductase 1 (CBR1) expression in human lung tissues and lung cancer cells by the cigarette smoke constituent benzo [a]pyrene. Toxicology Letters, 2012, 211(3): 266-273.

[13] SUTO K, KAJIHARA-KANO H, YOKOYAMA Y, HAYAKARI M, KIMURA J, KUMANO T, TAKAHATA T, KUDO H, TSUCHIDA S. Decreased expression of the peroxisomal bifunctional enzyme and carbonyl reductase in human hepatocellular carcinomas. Journal of Cancer Research and Clinical Oncology, 1999, 125(2): 83-88.

[14] UMEMOTO M, YOKOYAMA Y, SATO S, TSUCHIDA S, AL-MULLA F, SAITO Y. Carbonyl reductase as a significant predictor of survival and lymph node metastasis in epithelial ovarian cancer. British Journal of Cancer, 2001, 85(7): 1032-1036.

[15] TAKENAKA K, OGAWA E, OYANAGI H, WADA H, TANAKA F. Carbonyl reductase expression and its clinical significance in non-small-cell lung cancer. Cancer Epidemiology Biomarkers Prevention, 2005, 14(8): 1972-1975.

[16] ESPEY L L, YOSHIOKA S, RUSSELL D, UJIOKA T, VLADU B, SKELSEY M, FUJII S, OKAMURA H, RICHARDS J S. Characterization of ovarian carbonyl reductase gene expression during ovulation in the gonadotropin-primed immature rat. Biology of Reproduction, 2000, 62(2): 390-397.

[17] IWATA N, INAZU N, SATOH T. Changes and localization of ovarian carbonyl reductase during pseudopregnancy and pregnancy in rats. Biology of Reproduction, 1990, 43(3): 397-403.

[18] KACZMAREK M M, KRAWCZYNSKI K, BLITEK A, KIEWISZ J, SCHAMS D, ZIECIK A J. Seminal plasma affects prostaglandin synthesis in the porcine oviduct. Theriogenology, 2010, 74 (7): 1207-1220.

[19] WACLAWIK A, JABBOUR H N, BLITEK A, ZIECIK A J. Estradiol-17beta, prostaglandin E2 (PGE2), and the PGE2 receptor are involved in PGE2 positive feedback loop in the porcine endometrium. Endocrinology, 2009, 150(8): 3823-3832.

[20] WACLAWIK A, ZIECIK A J. Differential expression of prostaglandin (PG) synthesis enzymes in conceptus during peri-implantation period and endometrial expression of carbonyl reductase/PG 9-ketoreductase in the pig. Journal of Endocrinology, 2007, 194(3): 499-510.

[21] ZIECIK A J, WACLAWIK A, BOGACKI M. Conceptus signals for establishment and maintenance of pregnancy in pigs-lipid signaling system. Experimental and Clinical Endocrinology & Diabetes, 2008, 116(7): 443-449.

[22] WACLAWIK A, JABBOUR H N, BLITEK A, ZIECIK A J. Estradiol-17β, Prostaglandin E2 (PGE2), and the PGE2 receptor are involved in PGE2 positive feedback loop in the porcine endometrium. Endocrinology, 2009, 150(8): 3823-3832.

[23] FANG X, MOU Y, HUANG Z, LI Y, HAN L, ZHANG Y, FENG Y, CHEN Y, JIANG X, ZHAO W, SUN X, XIONG Z, YANG L, LIU H, FAN D, MAO L, REN L, LIU C, WANG J, LI K, WANG G, YANG S, LAI L, ZHANG G, LI Y, WANG J, BOLUND L, YANG H, WANG J, FENG S, LI S, DU Y. The sequence and analysis of a Chinese pig genome. Gigascience, 2012, 1(1): 16.

[24] 吴延博，陈从英，张志燕，郭源梅，高军. 猪精子黏合分子1(SPAM1)基因在白色杜洛克×二花脸F₂资源群体中的遗传变异及其与母猪产仔数的关联性. 中国农业科学, 2009, 42(6): 2111-2117.

WU Y B, CHEN C Y, ZHANG Z Y, GUO Y M, GAO J. Genetic variation of porcine sperm adhesion molecule 1 (SPAM1) gene and its association with litter size in a white duroc×erhualian resource population. Scientia Agricultura Sinica, 2009, 42(6): 2111-2117. (in Chinese)

[25] ONTERU S K, FAN B, DU Z Q, GARRICK D J, STALDER K J, ROTHSCHILD M F. A whole-genome association study for pig reproductive traits. Animal Genetics, 2012, 43(1):18-26.

[26] UIMARI P, SIRONEN A, SEVÓNAIMONEN M L. Whole-genome SNP association analysis of reproduction traits in the Finnish Landrace pig breed. Genetics Selection Evolution, 2011 43(1):1-8.

[27] TANAKA N, NONAKA T, NAKANISHI M, DEYASHIKI Y, HARA A, MITSUI Y. Crystal structure of the ternary complex of mouse lung carbonyl reductase at 1.8 A resolution: the structural origin of coenzyme specificity in the short-chain dehydrogenase/reductase family. Structure, 1996, 4(1):33-45.

[28] LINDEN M V D, BUCKINGHAM K, FARQUHAR C, KREMER J A M, METWALLY M. Luteal phase support for assisted reproduction cycles. Cochrane Database of Systematic Reviews, 2011, 10(10): CD009154.

[29] DEVOTO L, HENRÍQUEZ S, KOHEN P, STRAUSS J F. The significance of estradiol metabolites in human corpus luteum physiology. Steroids, 2017. 123:50-54.

[30] SANTOS V G, BEG M A, BETTENCOURT E M, GINTHER O J. Role of PGF2α in luteolysis based on inhibition of PGF2α synthesis in the mare. Theriogenology, 2013, 80(7): 812-820.

[31] ZHANG A L, SUN X Y, YIN Q, ZENG J H, ZHANG Z, LI J Q, ZHANG H. Functional characterization of the promoter of the carbonyl reductase1 (CBR1) gene in porcine endometrial cells. Journal of Zhejiang University Science B, 2017, 18(7): 626-634.

Related Articles 15

[1]	ZHAI XiaoHu,LI LingXu,CHEN XiaoZhu,JIANG HuaiDe,HE WeiHua,YAO DaWei. Quantitative Detection Technology of Porcine-Derived Materials in Meat by Real-time PCR [J]. Scientia Agricultura Sinica, 2023, 56(1): 156-164.
[2]	WANG YiDan,YANG FaLong,CHEN DiShi,XIANG Hua,REN YuPeng. One-Step Multiple TaqMan Real-time RT-PCR for Simultaneous Detection of Swine Diarrhea Viruses [J]. Scientia Agricultura Sinica, 2023, 56(1): 179-192.
[3]	YANG ShiMan, XU ChengZhi, XU BangFeng, WU YunPu, JIA YunHui, QIAO ChuanLing, CHEN HuaLan. Amino Acid of 225 in the HA Protein Affects the Pathogenicities of H1N1 Subtype Swine Influenza Viruses [J]. Scientia Agricultura Sinica, 2022, 55(4): 816-824.
[4]	ZHANG FengXi,XIAO Qi,ZHU JiaPing,YIN LiHong,ZHAO XiaLing,YAN MingShuai,XU JinHua,WEN LiBin,NIU JiaQiang,HE KongWang. Preparation and Identification of Monoclonal Antibodies to P30 Protein and Establishment of Blocking ELISA to Detecting Antibodies Against African Swine Fever Virus [J]. Scientia Agricultura Sinica, 2022, 55(16): 3256-3266.
[5]	WEI Tian,WANG ChengYu,WANG FengJie,LI ZhongPeng,ZHANG FangYu,ZHANG ShouFeng,HU RongLiang,LÜ LiLiang,WANG YongZhi. Preparation of Monoclonal Antibodies Against the p30 Protein of African Swine Fever Virus and Its Mapping of Linear Epitopes [J]. Scientia Agricultura Sinica, 2022, 55(15): 3062-3070.
[6]	ZHANG JingYuan,MIAO FaMing,CHEN Teng,LI Min,HU RongLiang. Development and Application of a Real-Time Fluorescent RPA Diagnostic Assay for African Swine Fever [J]. Scientia Agricultura Sinica, 2022, 55(1): 197-207.
[7]	Tao WANG,Yu HAN,Li PAN,Bing WANG,MaoWen SUN,Yi WANG,YuZi LUO,HuaJi QIU,Yuan SUN. Development of a TaqMan Real-Time PCR Targeting the MGF360-13L Gene of African Swine Fever Virus [J]. Scientia Agricultura Sinica, 2021, 54(5): 1073-1080.
[8]	JIA YunHui,XU ChengZhi,SUI JinYu,WU YunPu,XU BangFeng,CHEN Yan,YANG HuanLiang,QIAO ChuanLing,CHEN HuaLan. Immunogenicity Evaluation of Eukaryotic Expressing Plasmids Encoding HA Protein of Eurasian Avian-Like H1N1 Swine Influenza Virus [J]. Scientia Agricultura Sinica, 2019, 52(5): 930-938.
[9]	WANG Jia, ZENG ZhaoQiong, LIANG JianQiu, YU XiaoBo, WU HaiYing, ZHANG MingRong. Development New Molecular Markers for Quantitative Trait Locus (QTL) Analysis of the Seed Protein Content Based on Whole Genome Re-Sequencing in Soybean [J]. Scientia Agricultura Sinica, 2019, 52(16): 2743-2757.
[10]	Yuan SUN,HuaJi QIU. Eradication of Classical Swine Fever in China: Is It Far Away? [J]. Scientia Agricultura Sinica, 2018, 51(21): 4169-4176.
[11]	YuZi LUO,Yuan SUN,Tao WANG,HuaJi QIU. African Swine Fever: A Major Threat to the Chinese Swine Industry [J]. Scientia Agricultura Sinica, 2018, 51(21): 4177-4187.
[12]	CaiXia YIN, ShaoXiong YU, Kun SONG, Su LI, YuYing YANG, HuaJi QIU. The E2 Protein Mediates the Attachment and Internalization of Classical Swine Fever Virus in Cells [J]. Scientia Agricultura Sinica, 2018, 51(10): 1995-2003.
[13]	ZHANG Yu-jie, ZHAO Yan, XU Lu, ZHANG Qian-yi, CHEN Kai, SUN Yong-fang, ZOU Xing-qi, ZHU Yuan-yuan, ZHAO Qi-zu, NING Yi-bao, WANG Qin. Study of Location and Distribution of Classical Swine Fever Virus RNA in PK15 Cells by Visualization in Situ Hybridization Technology [J]. Scientia Agricultura Sinica, 2016, 49(12): 2397-2407.
[14]	JIN Xiu-zhe, HE Lei, CHENG Xiang-chao, ZHANG Chun-jie, YU Zu-hua, HAN Hai-feng, YANG Dan-fang, MEI Jing-jing, LI Yin-ju. Effect of Six Chinese Traditional Medicine Components on Inhibiting Swine Transmissible Gastroenteritis Virus in vitro [J]. Scientia Agricultura Sinica, 2016, 49(11): 2234-2244.
[15]	XU Hui-yang, XU Bang-feng, CHEN Yan, SUI Jin-yu, YANG Huan-liang, YIN Hang, YANG Da-wei, QIAO Chuan-ling, CHEN Hua-lan. Phylogenetic Analysis and Molecular Characteristics of an H1N1 Subtype Swine Influenza Virus [J]. Scientia Agricultura Sinica, 2015, 48(15): 3071-3078.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

The Cloning of Porcine CBR1 Gene’s Different Copy Forms and Analysis of Its Polymorphism

RichHTML

PDF