鸡SGOL2基因的克隆测序和表达分析

本研究通过对前期研究得到的候选鸟类卵巢特异表达基因LOC100857982进行克隆测序和表达分析,检验该基因是否在成年母鸡卵巢高度特异表达并对其功能进行初步分析,为进一步揭示卵巢的生长发育和成熟机制奠定理论基础。本研究采用RACE方法克隆出LOC100857982的cDNA全长,并进一步分析了该基因的序列,利用qRT-PCR方法对该基因进行表达分析。结果表明:LOC100857982的cDNA全长为1 306 bp;比对序列分析发现该基因是鸡SGOL2基因;系统进化树分析表明该基因与非洲爪蛙的遗传距离最近,符合基本的进化规律。qPCR结果显示该基因在25周龄白来航母鸡的卵巢中过表达。本研究表明鸡SGOL2基因是卵巢组织的特异表达基因。结合序列分析和表达分析,本研究推测该基因对母鸡的开产具有重要的作用。本研究还表明了实际克隆测序全长cDNA对基因功能研究的重要性,即使在高质量的鸡基因组中,Ensembl预测基因仍然有很多缺陷,从而导致以此为基础的一些生物学推断发生错误。

Molecular Cloning and Expression of Chicken SGOL2 Gene

Using molecular cloning and expression analysis,we focused on the investigation of LOC100857982 (SGOL2) gene which was forecast to be a bird specific gene expressed specifically in ovary of adult chicken.In order to study this gene,we obtained the full length cDNA(1 306 bp) of this gene by RACE technology.The results of phylogenetic tree of evolutionary relationship showed that chicken SGOL2 shared the closest evolutionary relationship with Xenopus SGOL2.The qRT-PCR showed that the expression in ovary of 25 weeks chicken was much higher than other stages of chicken development.Consequently,chicken SGOL2 gene may play an important role in the start of female reproduction.In chicken genome,there are some defects when using Ensembl predicted gene to infer species-specific gene.Some errors occurred in the evolutionary inference based on the predicted gene sequence.This study also indicated that cloning full length of cDNA is very important for gene function study.