牛Dmrt7基因的cDNA克隆及遗传变异  

Molecular Cloning and Genetic Variant of the Dmrt 7 gene in Cattle

作　　者：淮亚红[1] 许尚忠[2] 

机构地区：[1]中山火炬职业技术学院,广东中山528436 [2]中国农业科学院北京畜牧兽医研究所,北京100094

出　　处：《安徽农业科学》2009年第30期14607-14610,14615,共5页Journal of Anhui Agricultural Sciences

基　　金："十一五"国家科技支撑计划重大项目(2006BAD01A10);"十一五"国家高技术研究发展计划("863"计划)(2006AA10Z197)资助

摘　　要：[目的]探讨牛Dmrt7基因的生物学功能及群体遗传变异情况,为进一步研究该基因对牛精液品质的影响奠定基础。[方法]以牛睾丸组织为材料,根据GenBank上发表的小鼠的Dmrt7基因序列设计并合成2对引物,通过RT-PCR分别进行了扩增并运用DNA-MAN软件及在线工具对所得到的序列进行了生物信息学分析;同时,以牛肾脏、肝脏、睾丸、肺、脾脏、瘤胃、子宫、小肠、心脏、卵巢和肌肉共11个组织为材料,另外设计2对引物用于组织表达谱分析。利用PCR-SSCP技术和DNA测序对Dmrt7基因的多态性进行检测。[结果]获得了一个长为1616bp的cDNA片段(GenBank登录号为EF534775),该cDNA包含由1113个碱基组成的开放读码框(ORF),该ORF编码370个氨基酸;发现在第四内含子上有一个C/G突变,随后在277头本地牛品种和国外牛品种中进行群体多态性检测,发现G等位基因频率分布范围从0～0.4138;基因杂合度,有效等位基因数和多态信息含量分别为0～0.4851、1.0000～1.9423和0～0.3675。[结论]克隆了牛Dmrt7基因的cDNA序列并进行组织表达谱分析,该G等位基因在本地牛品种和外国牛品种中没有差异。[ Objective ] The paper aimed to study the biological function and group genetic variation situation of cattle Dmrt7 gene, which laid a foundation for the future study on the effects of this gene to the quality of cattle sperm . [ Method] Cattle testis tissues as materials, 2 pairs primers were designed and combined according to Dmrt7 gene sequence of mice published by GenBank, amplified through RT-PCR and sequence obtained from DNAMAN software and online tool were used for bioinformatics analysis; Meanwhile, 11 tissues of cattle kidney, liver, testis,lung, spleen, rumen, uterus, small intestine, heart, ovary, muscle as materials, besides, 2 pairs primers were designed for tissue expression profile analysis. Draft7 gene polymorphism were determined by PCR-SSCP technology and DNA sequence. [ Result] eDNA fragment of 1 616 bp length （ accession no is EF 534775）, including ORF composed of 1 113 bases , this ORF coded 370 amino acids; and a C/G mutation was found on the 4th intron, and then group polymorphism were tested in the 227 local cattle varieties and abroad cattle varieties, the distribution range of G allele frequency was 0-0. 413 8;the contents of gene heterozygosity, effective number of alleles and polymorphism information were 0 - 0.485 1,1. 000 0 - 1. 942 3, and 0 - 0. 367 5. [ Conclusion ] The eDNA sequence of cattle Dmrt7 gene was cloned and tissue expression profile were analyzed, there were no difference between local cattle variety and abroad cattle variety in G allele.

关 键 词：Dmrt7基因 遗传变异 牛 SNP 

分 类 号：S823.8[农业科学—畜牧学]