伏牛白山羊TACR1基因扩增及生物信息学分析

Amplification and Bioinformatics Analysis of TACR1 Gene in Funiu White Goat

本研究旨在克隆伏牛白山羊TACR1基因,并对其结构和编码蛋白进行生物信息学分析,为进一步研究TACR1基因的功能及其在生殖生理和免疫调节中的作用奠定基础。根据GenBank中已发表的山羊TACR1基因(登录号:NC_030818.1)序列设计引物,对伏牛白山羊TACR1基因进行分段PCR扩增并测序,拼接后得到包含TACR1基因完整编码区的序列片段。结果表明,伏牛白山羊TACR1基因开放阅读框(ORF)全长852bp,共编码283个氨基酸,碱基组成为:A(23.47%)、T(25.49%)、C(27.76%)和G(23.27%)。与绵羊相比,伏牛白山羊TACR1基因编码区发生了2个碱基突变,但没有引起氨基酸的改变。5′端非编码序列长为860bp,发生了9个碱基突变;3′端非编码序列长为271bp,发生了2个碱基突变。同源性分析结果显示,伏牛白山羊与卡普拉山羊、藏羚羊、绵羊、瘤牛、白尾鹿、白鳍豚、人、野猪、金钱豹、土拨鼠、野驴TACR1基因同源性分别为99.9%、99.3%、99.2%、97.5%、96.7%、92.9%、89.7%、88.6%、86.3%、86.2%和85.0%。SignalP 4.1在线软件预测结果显示,TACR1N末端存在信号肽且可能在21位氨基酸处存在裂解位点;蛋白质结构预测发现伏牛白山羊TACR1基因编码蛋白没有跨膜结构域。

The purpose of this study was to clone the TACR 1 gene of Funiu White goat and analyze its structure and encoded protein by bioinformatics methods to lay a foundation for further study on the function of TACR 1 gene and its role in reproductive physiology and immune regulation.5 pairs of primers were designed based on the goat TACR 1 gene sequence in GenBank(accession No:NC_030818.1).The Funiu White goat TACR 1 gene was amplified by PCR amplification and sequenced,and the complete coding region(CDS)of TACR 1 gene was obtained after splicing the fragments.The results showed that the open reading frame(CRF)of TACR 1 gene in Funiu White goat was 852 bp in length and encoded a total of 283 amino acids.The content of base composition was A(23.47%),T(25.49%),C(27.76%)and G(23.27%).Compared with sheep TACR 1 gene,the TACR 1 gene CDS region of the Funiu White goat had a two-base mutation which did not cause amino acid changes.The non-coding region at the 5′and 3′ends was 860 and 271 bp in length,and had 9 and 2 base mutations,respectively.The results of homology analysis showed that the identity of TACR 1 gene between Funiu White goat and Capra hircus,Pantholops hodgsonii,Ovis aries,Bos indicus,Odocoileus virginianus,Lipotes vexillifer,Homo sapiens,Sus scrofa,Panthera pardus,Cavia porcellus,Equus asinus were 99.9%,99.3%,99.2%,97.5%,96.7%,92.9%,89.7%,88.6%,86.3%,86.2%and 85.0%,respectively.There was a signal peptide at the N-terminus of TACR1 and there might be a cleavage site at 21th amino acid predicted by SignalP4.1 online software.Through the prediction of protein structure,it was found that the protein encoded by TACR 1 gene of the FuNiu White goat had no transmembrane domain.