半滑舌鳎膜孕激素受体基因克隆与组织表达分析  

Molecular cloning and tissue expression analysis of membrane progestin receptor alpha gene(mPRα)from half smooth tongue sole Cynoglossus semilaevis Günther

作　　者：史宝[1] 李晓晓[1] 柳学周[1] 徐永江[1] 王珊珊[1] 刘芝亮[1] 王妍妍[1] 

机构地区：[1]农业部海洋渔业可持续发展重点实验室青岛市海水鱼类种子工程与生物技术重点实验室中国水产科学研究院黄海水产研究所,266071

出　　处：《渔业科学进展》2013年第3期61-67,共7页Progress in Fishery Sciences

基　　金：国家鲆鲽类产业技术体系(CARS-50);国家自然科学基金(31201982);公益性农业行业专项项目(200903005);留学人员科技活动项目择优资助经费共同资助

摘　　要：采用同源克隆和末端快速扩增(RACE)方法,首次获得全长为1319bp的半滑舌鳎膜孕激素受体(mPRα)的cDNA序列;将推断的氨基酸序列与其他物种mPRα氨基酸序列进行多重比较分析,发现存在7个跨膜区域。使用MEGA4.0临位相联法和ClustalX方法对mPRα的氨基酸序列进行聚类分析和序列相似度分析。结果表明,半滑舌鳎mPRα与漠斑牙鲆、大西洋绒须石首鱼以及青鳉等鱼类的mPRα聚为一支,亲缘关系较近,相似度较高,分别为94%、93%和90%;而与高等哺乳动物人和牛相似性均为53%,亲缘关系较远。应用半定量RT-PCR技术分析了mPRαmRNA在性成熟雌性半滑舌鳎不同组织的表达,结果表明,mPRαmRNA组织表达具有广泛性,但表达量存在差异,在脑、肾、脾和卵巢等组织表达丰富,在肝、胃和肌肉组织表达较弱。Full-length cDNA encoding membrane progestin receptor alpha gene（mPRα） was firstly cloned from half smooth tongue sole Cynoglossus semilaevis Günther by homology cloning and RACE-PCR analysis.The length of complete cDNA sequence of mPRαgene was 1 319bp.Sequence alignment of deduced amino acid of tongue sole mPRαand amino acid of other species showed that there were seven transmembrane domains.The rooted phylogenetic tree was constructed by the neighbor-joining method of MEGA 4.0,and the identity of tonguesole mPRαwith other representative sequences was analyzed by ClustalX.The results indicated that the tongue sole mPRαwas clustered together with mPRαof other fish.The identity was 94%,93%and 90% when compared with Southern flounder Paralichthys lethostigma,Atlantic croaker Micropogonias undulates and Japanese medaka Oryziaslatipes,respectively.In contrast,identity among the mPRαof tongue sole and that of human Homo sapiens and cattle Bos taurus was low,only 53%.A semi-quantitative RT-PCR was developed to measure mRNA expression levels of mPRαgene of female tongue sole.Tissue expression analysis showed that mPRαmRNA was expressed widely in tongue sole,although the expression level was not homogeneous.mPRαtranscripts were highly abundant in brain,kidney,spleen and ovary,less abundant in liver,stomach and muscle.

关 键 词：半滑舌鳎 膜孕激素受体基因 基因克隆 组织表达 

分 类 号：S917.4[农业科学—水产科学]