Molecular cloning and genetic variant of the Dmrt 7 gene in cattle | 牛Dmrt7基因的cDNA克隆及遗传变异

Chinese. ［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DNAMAN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1 616 bp的cDNA 片段（GenBank 登录号为EF 534775 ），该 cDNA 包含由1 113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0~0.413 8；基因杂合度，有效等位基因数和多态信息含量分别为0~0.485 1、1.000 0~1.942 3和0~0_367 5。 ［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。

English. ［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. Dmrt7 gene polymorphism were determined by PCR-SSCP technology and DNA sequence. ［Result］cDNA 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 cDNA 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. _