云南农业大学学报(自然科学)
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云南农业大学学报(自然科学) 2017, Vol. 32 Issue (4) :612-619    DOI: 10.16211/j.issn.1004-390X(n).2017.04.008
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槟榔江水牛Dmrt7基因克隆及分子特征、组织表达谱分析
伍姗1, 吉丽2, 张永云3, 杜平4, 康定富4, 哈福1, 苗永旺1
1. 云南农业大学 动物科学技术学院, 云南 昆明 650201;
2. 云南农业职业技术学院 畜牧兽医学院, 云南 昆明 650212;
3. 云南农业大学 农科专业基础实验教学示范中心, 云南 昆明 650201;
4. 腾冲市巴福乐槟榔江水牛良种繁育有限公司, 云南 腾冲 679118
Cloning,Molecular Characterization and Tissue Expression Profile of Binglangjiang Buffalo Gene Dmrt7
WU Shan1, JI Li2, ZHANG Yongyun3, DU Ping4, KANG Dingfu4, HA Fu1, MIAO Yongwang1
1. Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;
2. Department of Animal Husbandry & Veterinary Medicine, Yunnan Agricultural College of Vocational Education, Kunming 650212, China;
3. Teaching Demonstration Center of the Basic Experiments of Agricultural Majors, Yunnan Agricultural University, Kunming 650201, China;
4. Tengchong Co., Ltd. of Fine Variety Breeding of Binglangjiang Buffalo, Tengchong 679118, China

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摘要 

[目的]Dmrt7基因是Dmrt基因家族成员之一,影响公畜精子形成。迄今,有关水牛精子发生的遗传基础的研究很少,本研究旨在对槟榔江水牛Dmrt7基因完整CDS区进行克隆和组织差异表达分析。[方法]采用RT-PCR和功能生物信息学方法。[结果]槟榔江水牛Dmrt7基因完整编码区序列长1 113 bp,编码370个氨基酸;Dmrt7包含DM和DMRT-like两个保守结构域,无信号肽,无跨膜结构,为核内蛋白。同源性分析显示,水牛与其他牛科物种Dmrt7氨基酸序列相似度达95%以上。在检测的10个组织中,Dmrt7基因仅在水牛睾丸组织中表达且高表达。在第6外显子发现1个SNP位点(c.665A>G),引起了p.Q222R的替换,此位点的替换对Dmrt7蛋白功能无影响。[结论]有助于了解水牛Dmrt7基因的基本功能。

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伍姗
吉丽
张永云
杜平
康定富
哈福
苗永旺
关键词槟榔江水牛   Dmrt7基因   DM结构域   精子生成   遗传变异   生物信息学分析     
Abstract

[Purposes] As one of the members of Dmrt gene family, Dmrt7 gene affects the spermiogenesis of male animals. So far, few studies have been done related to the genetic basis of buffalo spermiogenesis. In this study, by employed Binglangjiang water buffalo as research object, the complete CDS sequence of water buffalo Dmrt7 gene was cloned, and its sequence characterization, tissue differential expression were assayed and analysed.[Methods] RT-PCR and functional bioinformatics methods were used.[Results] The results showed that the CDS of Binglangjiang buffalo Dmrt7 gene is 1113 bp in length, which encodes a polypeptide composed of 370 amino acids. Buffalo Dmrt7 contains two conserved domains, viz. a DM and a DMRT-like domain. This protein was predicted a nuclear protein without signal peptide and transmembrane domain. Homology analysis showed that the amino acid sequence similarity of Dmrt7 between buffalo and other bovine species was more than 95%. Among 10 different tissues of adult buffalo, Dmrt7 gene was detected to only express in the testis of water buffalo, and its expression level was dominantly high. The polymorphism c.665A>G was found in exon 6 of CDS of buffalo Dmrt7 gene which leads to a p.Q222R change. But this substitution was presumed to have no functional effect on buffalo Dmrt7 protein.[Conclusion] The results of this study will provide fundamental insights into the function of buffalo Dmrt7 gene.

KeywordsBinglangjiang buffalo   Dmrt7 gene   DM domain   spermatogenesis   expression profile   bioinformatics analysis     
Received 2017-01-31;
Fund:

国家自然科学基金项目(31460582,30660024);云南省应用基础研究重点项目(2014FA032,2007C0003Z);云南省应用基础研究计划面上项目(2006C0034M,2010ZC243)。

Corresponding Authors: 哈福(1974-),男,宁夏银川人,博士,副教授,主要从事动物科学研究。E-mail:119272386@qq.com;苗永旺(1964-),男,内蒙古通辽人,博士,教授,博士生导师,主要从事动物遗传学研究。E-mail:yongwangmiao1@126.com     Email: 119272386@qq.com;yongwangmiao1@126.com
About author: 伍姗(1992-),女,湖南常德人,在读硕士研究生,主要从事动物分子遗传学研究。E-mail:610319340@qq.com;吉丽(1971-),女,云南石屏人,硕士,讲师,主要从事动物科学领域的教学、科研工作。E-mail:hjkm71524@126.com
引用本文:   
伍姗, 吉丽, 张永云, 杜平, 康定富, 哈福, 苗永旺.槟榔江水牛Dmrt7基因克隆及分子特征、组织表达谱分析[J]  云南农业大学学报(自然科学), 2017,V32(4): 612-619
WU Shan, JI Li, ZHANG Yongyun, DU Ping, KANG Dingfu, HA Fu, MIAO Yongwang.Cloning,Molecular Characterization and Tissue Expression Profile of Binglangjiang Buffalo Gene Dmrt7[J]  Journal of Yunnan Agricultural University, 2017,V32(4): 612-619
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