Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (5): 1036-1043.doi: 10.3864/j.issn.0578-1752.2013.05.019

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Sequence Analysis and Study on the Expression Level of Dmrt7 Gene in Yak and Cattle-Yak Testis

 JIN  Shuai, GUO  Xian, BAO  Peng-Jia, LIANG  Chun-Nian, WU  Xiao-Yun, LIU  Jian, YAN  Ping   

  1. 1.Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences/Key Laboratory of Yak Breeding Engineering, Lanzhou 730050
    2.School of Basic Medicine, Lanzhou University, Lanzhou 730030
  • Received:2012-08-27 Online:2013-03-01 Published:2012-10-19

PDF

579

Abstract: 【Objective】The aim of the study is to investigate the coding region sequences, their structures and the expression levels of Dmrt7 gene in yak and cattle-yak testis, for offering references for the revealing of the molecular mechanism of infertility of cattle-yak.【Method】A coding region sequence of yak and cattle-yak Dmrt7 was cloned by molecular cloning technique in this study. Some characters of the Dmrt7 gene and encoded protein sequences were predicted and analyzed by the methods of bioinformatics in the following aspects as the functional sites and secondary structure. The expression levels of Dmrt7 mRNA in some organs of yak were detected by semi-quantitative PCR. Real-time PCR was employed to examine the expression levels of Dmrt7 mRNA in yak and cattle-yak testis. Western blotting was applied to detect the expression levels of Dmrt7 protein in yak and cattle-yak testis. 【Result】 The coding region sequences of Dmrt7 gene in yak and cattle-yak were consistent，contains a complete ORF (1 113 bp) which encoding 370 amino acids. The protein encoded by yak and cattle-yak Dmrt7 contains a DM domain. The secondary structures were mainly composed of Helix, Coli and Extended strand. Dmrt7 mRNA expression in the testis was detected, but not in any other tissue tested. And the difference of expression level of Dmrt7 mRNA and protein in testis between yak and cattle-yak was extremely remarkable (P＜0.01).【Conclusion】The expression levels of Dmrt7 mRNA and protein in yak testis were significantly higher than cattle-yak testis，and the expression levels of Dmrt7 mRNA and protein were consistent. The results indicate that the expression level of Dmrt7 protein in cattle-yak may be too low to block the meiosis process.

Key words: yak , cattle-yak , testis , Dmrt7 , cloning , bioinformaticanalysis , expression level

[1]屈旭光, 李齐发, 刘振山, 董丽艳, 李新福, 郝称莉, 谢庄. 牦牛、犏牛睾丸组织中SYCP3基因mRNA 表达水平研究. 畜牧兽医学报, 2008, 39(8): 1132-1136.

Qu X G, Li Q F, Liu Z S, Dong L Y, Li X F, Hao C L, Xie Z. Study on the expression level of SYCP3 mRNA in yak, and cattle-yak testis. Acta Veterinaria et Zootechnica Sinica, 2008, 39(8): 1132-1136. (in Chinese)

[2]Zhang Q, Li J, Li Q, Li X, Liu Z, Song D, Xie Z. Cloning and characterization of the gene encoding the bovine BOULE protein.  Molecular Genetics and Genomics, 2009, 281(1): 67-75.

[3]Erdman S E, Burtis K C. The Drosophila doublesex proteins share a novel zinc finger related DNA binding domain. The Embo Journal, 1993, 12(2): 527-535.

[4]Raymond C S, Shamu C E, Shen M M, Seifert K J, Hirsch B, Hodgkin J, Zarkower D. Evidence for evolutionary conservation of sex-determining genes. Nature, 1998, 391(6668): 691-695.

[5]Zarkower D. Establishing sexual dimorphism: conservation amidst diversity? Nature Reviews Genetics, 2001, 2(3): 175-185.

[6]Lints R, Emmons S W. Regulation of sex-specific differentiation and mating behavior in C. elegans by a new member of the DM domain transcription factor family. Genes Development, 2002, 16(18): 2390-2402.

[7]Kondo M, Froschauer A, Kitano A, Nanda I, Hornung U, Volff J N, Asakawa S, Mitani H, Naruse K, Tanaka M, Schmid M, Shimizu N, Schartl M, Shima A. Molecular cloning and characterization of DMRT genes from the medaka oryzias latipes and the platyfish xiphophorus maculatus. Gene, 2002, 295(2): 213-222.

[8]Volff J N, Zarkower D, Bardwell V J, Schartl M. Evolutionary dynamics of the DM domain gene family in metazoans. Journal of Molecular Evolution, 2003, 57(Suppl.1): S241-S249.

[9]Brunner B, Hornung U, Shan Z, Nanda I, Kondo M, Zend-Ajusch E, Haaf T, Ropers H H, Shima A, Schmid M, Kalscheuer V M, Schartl M. Genomic organization and expression of the doublesex-related gene cluster in vertebrates and detection of putative regulatory regions for DMRT1. Genomics, 2001, 77(1/2): 8-17.

[10]Ottolenghi C, Fellous M, Barbieri M, McElreavey K. Novel paralogy relations among human chromosomes support a link between the phylogeny of doublesex-related genes and the evolution of sex determination. Genomics, 2002, 79(3): 333-343.

[11]Kim S, Kettlewell J R, Anderson R C, Bardwell V J, Zarkower D. Sexually dimorphic expression of multiple doublesex-related genes in the embryonic mouse gonad. Gene Expression Patterns, 2003, 3(1): 77-82.

[12]Veith A M, Klattig J, Dettai A, Schmidt C, Englert C, Volff J N. Male-biased expression of X-chromosomal DM domain-less Dmrt8 genes in the mouse. Genomics, 2006, 88(2): 185-195.

[13]Heard E, Disteche C M. Dosage compensation in mammals: fine-tuning the expression of the X chromosome. Genes Development, 2006, 20(14): 1848-1867.

[14]Huynh K D, Lee J T. X-chromosome inactivation: a hypothesis linking ontogeny and phylogeny. Nature Reviews Genetics, 2005, 6(5): 410-418.

[15]McKee B D, Handel M A. Sex chromosomes, recombination, and chromatin conformation. Chromosoma, 1993, 102(2): 71-80.

[16]Mahadevaiah S K, Turner J M, Baudat F, Rogakou E P, de Boer P, Blanco-Rodríguez J, Jasin M, Keeney S, Bonner W M, Burgoyne P S. Recombinational DNA double-strand breaks in mice precede synapsis. Nature Genetics, 2001, 27(3): 271-276.

[17]Jablonka E, Lamb M J. Meiotic pairing constraints and the activity of sex chromosomes. Journal of Theoretical Biology, 1988, 133(1): 23-36.

[18]Odorisio T, Rodriguez T A, Evans E P, Clarke A R, Burgoyne P S. The meiotic checkpoint monitoring synapsis eliminates spermatocytes via p53-independent apoptosis. Nature Genetics, 1998, 18(3): 257-261.

[19]Hoyer-Fender S. Molecular aspects of XY body formation. Cytogenetic and Genome Research, 2003, 103(3/4): 245-255.

[20]Fernandez-Capetillo O, Mahadevaiah S K, Celeste A, Romanienko P J, Camerini-Otero R D, Bonner W M, Manova K, Burgoyne P, Nussenzweig A. H2AX is required for chromatin remodeling and inactivation of sex chromosomes in male mouse meiosis. Development Cell, 2003, 4(4): 497-508.

[21]Kim S, Namekawa S H, Niswander L M, Ward J O, Lee J T, Bardwell V J, Zarkower D. A mammal-specific doublesex homolog associates with male sex chromatin and is required for male meiosis. PLoS Genetics, 2007, 3(4): e62.

[22]Kawamata M, Nishimori K. Mice deficient in Dmrt7 show infertility with spermatogenic arrest at pachytene stage. FEBS Letters, 2006, 580(27): 6442-6446.

[23]胡欧明, 钟金城, 陈智华, 马力, 蔡立. 犏牛精母细胞联会复合体与雄性不育关系的研究. 西南民族学院学报: 自然科学版, 2000, 26(1): 61-66.

Hu O M, Zhong J C, Chen Z H, Ma L, Cai L. A study on relationship between synaptonemal complex and male sterility of pian niu. Journal of Southwest Nationalities College: Natural Science Edition, 2000, 26(1): 61-66. (in Chinese)

[24]董丽艳, 李齐发, 屈旭光, 李隐侠, 李新福, 徐洪涛, 谢庄. 黄牛、牦牛和犏牛睾丸组织中Cdc2、Cdc25A 基因mRNA 表达水平. 遗传, 2009, 31(5): 495-499.

Dong L Y, Li Q F, Qu X G, Li Y X, Li X F, Xu H T, Xie Z. Expression levels of Cdc2 and Cdc25A mRNA in cattle, yak, and cattle-yak testis. Hereditas, 2009, 31(5): 495-499. (in Chinese)

[25]李贤, 李齐发, 赵兴波, 徐洪涛, 顾垚, 朱翔, 谢庄, 刘红林. 牦牛和犏牛Dmc1基因序列分析及睾丸组织转录水平研究. 中国农业科学, 2010, 43(15): 3221-3229.

Li X, Li Q F, Zhao X B, Xu H T, Gu Y, Zhu X, Xie Z, Liu H L. Sequence analysis and study on the expression level of Dmc1 mRNA in yak and cattle-yak testis. Scientia Agricultura Sinica, 2010, 43(15): 3221-3229. (in Chinese)

[26]潘增祥, 刘振山, 李隐侠, 于莎莉, 李明桂, 谢庄, 李齐发. 犏牛及其亲本睾丸组织中印记基因SNRPN DMR甲基化与mRNA 表达差异研究. 中国农业科学, 2010, 43(22): 4709-4716.

Pan Z X, Liu Z S, Li Y X, Yu S L, Li M G, Xie Z, Li Q F. Difference of SNRPN methylation status and its mRNA expression in testes between cattle-yaks and their parents. Scientia Agricultura Sinica, 2010, 43(22): 4709-4716. (in Chinese)

[27]李明桂, 徐洪涛, 李隐侠, 于莎莉, 赵兴波, 潘增祥, 朱翔, 谢庄, 李齐发. b-Boule基因5′调控序列的克隆与睾丸组织DMR甲基化分析. 中国农业科学, 2011, 44(18): 3859 -3867.

Li M G, Xu H T, Li Y X, Yu S L, Zhao X B, Pan Z X, Zhu X, Xie Z, Li Q F. Cloning and testicular DMR methylation analysis of b-boule gene 5′-flanking regulation regions. Scientia Agricultura Sinica, 2011, 44(18): 3859-3867. (in Chinese)

[28]Cohen P E, Pollack S E, Pollard J W. Genetic analysis of chromosome pairing, recombination, and cell cycle control during first meiotic prophase in mammals. Endocrine Reviews, 2006, 27(4): 398-426.

[29]Roeder G S, Bailis J M. The pachytene checkpoint. Trends Genet, 2000, 16(9): 395-403.

[30]Hou Y, Zhou X, Liu J, Yuan J, Cheng H, Zhou R. Nuclear factor-Y (NF-Y) regulates transcription of mouse Dmrt7 gene by binding to tandem CCAAT boxes in its proximal promoter. International Journal of Biological Sciences, 2010, 6(7): 655-664.
[1] GU LiDan,LIU Yang,LI FangXiang,CHENG WeiNing. Cloning of Small Heat Shock Protein Gene Hsp21.9 in Sitodiplosis mosellana and Its Expression Characteristics During Diapause and Under Temperature Stresses [J]. Scientia Agricultura Sinica, 2023, 56(1): 79-89.
[2] WANG JiaMin,SHI JiaChen,MA FangFang,CAI Yong,QIAO ZiLin. Effects of Soy Isoflavones on the Proliferation and Apoptosis of Yak Ovarian Granulosa Cells [J]. Scientia Agricultura Sinica, 2022, 55(8): 1667-1675.
[3] LI YuZe,ZHU JiaWei,LIN Wei,LAN MoYing,XIA LiMing,ZHANG YiLi,LUO Cong,HUANG Gui Xiang,HE XinHua. Cloning and Interaction Protein Screening of RHF2A Gene from Xiangshui Lemon [J]. Scientia Agricultura Sinica, 2022, 55(24): 4912-4926.
[4] DUAN YaRu,GAO MeiLing,GUO Yu,LIANG XiaoXue,LIU XiuJie,XU HongGuo,LIU JiXiu,GAO Yue,LUAN Feishi. Map-Based Cloning and Molecular Marker Development of Watermelon Fruit Shape Gene [J]. Scientia Agricultura Sinica, 2022, 55(14): 2812-2824.
[5] QU Cheng,WANG Ran,LI FengQi,LUO Chen. Cloning and Expression Profiling of Gustatory Receptor Genes BtabGR1 and BtabGR2 in Bemisia tabaci [J]. Scientia Agricultura Sinica, 2022, 55(13): 2552-2561.
[6] RAN HongBiao,ZHAO LiLing,WANG Hui,CHAI ZhiXin,WANG JiKun,WANG JiaBo,WU ZhiJuan,ZHONG JinCheng. Effects of lncFAM200B on the Lipid Deposition in Intramuscular Preadipocytes of Yak [J]. Scientia Agricultura Sinica, 2022, 55(13): 2654-2666.
[7] ZHANG Li,ZHANG Nan,JIANG HuQiang,WU Fan,LI HongLiang. Molecular Cloning and Expression Pattern Analysis of NPC2 Gene Family of Apis cerana cerana [J]. Scientia Agricultura Sinica, 2022, 55(12): 2461-2471.
[8] ZHANG Lu,ZONG YaQi,XU WeiHua,HAN Lei,SUN ZhenYu,CHEN ZhaoHui,CHEN SongLi,ZHANG Kai,CHENG JieShan,TANG MeiLing,ZHANG HongXia,SONG ZhiZhong. Identification, Cloning, and Expression Characteristics Analysis of Fe-S Cluster Assembly Genes in Grape [J]. Scientia Agricultura Sinica, 2021, 54(23): 5068-5082.
[9] TAN YongAn,JIANG YiPing,ZHAO Jing,XIAO LiuBin. Expression Profile of G Protein-Coupled Receptor Kinase 2 Gene (AlGRK2) and Its Function in the Development of Apolygus lucorum [J]. Scientia Agricultura Sinica, 2021, 54(22): 4813-4825.
[10] LI ZiTeng,CAO YuHan,LI Nan,MENG XiangLong,HU TongLe,WANG ShuTong,WANG YaNan,CAO KeQiang. Molecular Variation and Phylogenetic Relationship of Apple Scar Skin Viroid in Seven Cultivars of Apple [J]. Scientia Agricultura Sinica, 2021, 54(20): 4326-4336.
[11] WANG Na,ZHAO ZiBo,GAO Qiong,HE ShouPu,MA ChenHui,PENG Zhen,DU XiongMing. Cloning and Functional Analysis of Salt Stress Response Gene GhPEAMT1 in Upland Cotton [J]. Scientia Agricultura Sinica, 2021, 54(2): 248-260.
[12] TAN YongAn,ZHAO XuDong,JIANG YiPing,ZHAO Jing,XIAO LiuBin,HAO DeJun. Cloning, Preparation of Antibody and Response Induced by 20-Hydroxyecdysone of Target of Rapamycin in Apolygus lucorum [J]. Scientia Agricultura Sinica, 2021, 54(10): 2118-2131.
[13] LI XueRu,SHI XiXiong,WANG JianZhong,ZHANG PanGao,TIAN Zhu,HAN Ling. Effect of Nitric Oxide Synthetase Inhibitor on Yak Meat Quality During Post-Mortem Aging [J]. Scientia Agricultura Sinica, 2020, 53(8): 1617-1626.
[14] KunNeng ZHOU,JiaFa XIA,Peng YUN,YuanLei WANG,TingChen MA,CaiJuan ZHANG,ZeFu LI. Transcriptome Research of Erect and Short Panicle Mutant esp in Rice [J]. Scientia Agricultura Sinica, 2020, 53(6): 1081-1094.
[15] MIAO JianJun,PENG ZhongLi,GAO YanHua,BAI Xue,XIE XinTing. Effects of Dietary Small Peptides on Production Performance and Expression of PepT1 mRNA in Digestive Tract of Fattening Yaks [J]. Scientia Agricultura Sinica, 2020, 53(23): 4950-4960.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd