版纳微型猪近交系TSARG7基因克隆及功能生物信息学分析

Cloning and Functional Bioinformatics Analysis of TSARG7 Gene in Banna Mini-pig Inbred Line

为获得版纳微型猪近交系(Banna Mini-pig inbred line,BMI)TSATG7基因序列并分析组织表达情况,本研究以GenBank中猪及近缘物种的TSARG7基因mRNA序列为参考序列,设计特异引物扩增BMI TSARG7基因,半定量分析10个重要组织的表达谱,并对蛋白质序列进行功能生物信息学分析。结果显示,获得了BMI TSARG7基因1 371bp的编码区序列(GenBank登录号:KU950831,对应的氨基酸登录号:AMY60405),编码456个氨基酸,蛋白质分子质量为52.05ku,等电点(pI)为9.28。多组织表达分析表明,TSARG7基因在睾丸中高表达,在其他组织中呈中、低表达。功能生物信息学分析表明,TSARG7蛋白质存在1个保守结构域,有3个跨膜螺旋结构,无信号肽序列;N末端疏水,C末端亲水;有5类功能活性位点,位于细胞质的概率是94.1%。本试验结果为进一步研究TSARG7基因在猪精子发生和性成熟方面的作用及功能奠定基础。

In order to obtain TSATG7 gene sequence and analyze mRNA expression in tissue of Banna Mini-pig inbred line （BMI）, we used TSARG7 gene mRNA sequence of pig and related species from GenBank as reference sequences to design specific primers and amplify BMI TSARG7 gene. The semi-quantitative was used to analyze the expression of TSARG7 gene in 10 important tissues, protein sequence was used to carry out functional bioinformatics analysis. A coding se- quence of 1 371 bp （GenBank accession No. ： KU950831, the corresponding amino acid sequence accession No. ： AMY60405） of BMI TSARG7 was obtained, which encoded a protein of 456 ami- no acids, the protein molecular weight was 52.05 ku, and the isoelectric point was 9.28. Multiple tissue expression indicated that TSARG7 gene expressed highly in the testis, middle and low ex- pression in the other tissues. Functional bioinformatics analysis indicated that TSARG7 proteincontained one conserved domain, three transmembrane regions, and no signal peptide sequences; Its N-terminal was hydrophobic and C-terminal was hydrophilic;It had five kinds functional active sites and a 94.1 % to located in cytoplasmic. The results of the study would lay a foundation for further study of the gene about its functions in pig spermatogenesis and sexual maturation.