4个不同地理种群中华鳖POMC基因全长cDNA克隆及其序列分析

为探讨中华鳖不同地理种群的POMC基因的多态性,建立区分中华鳖不同地理种群的分子遗传标记,采用RT-PCR的方法扩增得到4个不同地理种群(太湖鳖种群、沙鳖种群、台湾鳖种群和黄河鳖种群)中华鳖的POMC基因,对该基因进行克隆及序列测定,并对所测序列进行比较,分析其SNP位点,并对所得的SNP位点进行验证。结果显示:4个不同地理种群中华鳖所扩增的POMC基因的核苷酸序列均为786bp;以megalign软件比较分析4个不同地理种群中华鳖的POMC基因,发现该序列中共有15个多态性核苷酸位点,经过验证,第129位核苷酸的变异为台湾鳖种群不同于其他种群的变异,第441位核苷酸变异为太湖鳖种群不同于其他种群的变异,第168和396位核苷酸为黄河鳖种群不同于其他种群的变异,第531和618位核苷酸为沙鳖种群不同于其他种群的变异。结果表明,中华鳖的POMC基因存在多态性,不同地理种群中华鳖POMC基因均有各自特异的核苷酸序列,可以作为区分中华鳖的不同地理种群的分子遗传标记。

Clone and polymorphism of POMC gene in four populations of pelodoiscus sinensis

To study the polymorphism of the proopiomelanocortin(POMC) gene and establish the molecular genetic marker for identifying different population of Pelodoiscus sinensis,populations from Taihu Lake,Huanghe River,Taiwan-introduced and Pelodiscus axenaria,were collected to get the blood.The POMC gene was amplified by RT-PCR,sequenced and analyzed thereafter.Fifteen SNPs were detected in the PCR-amplified 786 bp fragments.One SNP at nucleotide 129 in Taiwan-introduced population,one SNP at nucleotide 441 in Taihu Lake population,two SNPs at nucleotide 168 and 396 in Huanghe River population,and two SNPs at nucleotide 531 and 618 in P.axenaria were identified.The results showed that the POMC gene of P.sinensis showed polymorphic,the POMC gene of different population has specific mutation site,which can be used as a molecular genetic marker for identifying different population of P.sinensis.