鲤鱼生长激素在毕赤酵母中的表达  

Expression of Common Carp Growth Hormone in Yeast P. pastoris

作　　者：李英华[1] 白俊杰[2] 李新辉[2] 叶星[2] 简清[2] 罗建仁[2] 梁旭方[1] 

机构地区：[1]暨南大学生命科学技术学院生物工程系,广州510632 [2]中国水产科学研究院珠江水产研究所农业部热带亚热带鱼类选育与养殖重点开放实验室,广州510380

出　　处：《中国生物化学与分子生物学报》2001年第4期488-491,共4页Chinese Journal of Biochemistry and Molecular Biology

基　　金：农业部"九五"渔业科技重点项目 (No .95 B 96 0 2 0 2 0 4);中国水产科学研究院重点基金项目 (No .99 0 8 0 2 )&&

摘　　要：将编码鲤鱼 (Cyprinuscarpio )生长激素 (GH)成熟肽的cDNA克隆到毕赤酵母 (P .pastoris)胞内表达载体pHIL D2中 ,构建重组表达质粒pHIL D2 GH .转化组氨酸缺陷型酵母GS115,获得表达鲤鱼GH的酵母工程菌 .经甲醇诱导 ,SDS PAGE和Western印迹检测表明 ,鲤鱼GH在酵母中得到表达 ,表达产物在胞内以可溶状态存在 ,具有鲤鱼GH的免疫活性 .用诱导后的酵母投喂罗非鱼 。Common carp ( Cyprinus carpio ) growth hormone cDNA was cloned into the methylotrophic yeast P. pastoris integrative expression vector pHIL D2, under the control of the alcohol oxidase promoter. The lineanized recombinant plasmid was then transformed into HIS4 mutant yeast GS115 by electroporation. PCR detection method was used to screen the positive transformants and MM medium was used for further selection of the Mut + transformants. After being induced by methanol, the yeast cell was collected and broken by ultrasonic processor.In the supernatant of the yeast lysates, a specific 22 kD protein band appeared in SDS PAGE. The immunological activity of the GH was confirmed by Western blotting. The yeast containing recombinant common carp growth hormone was applied as supplement to feed Tilapia fingerling. The result showed that the recombinant GH had significant growth promoting effects on Tilapia.

关 键 词：鲤鱼生长激素 毕赤酵母 胞内表达 促生长作用 

分 类 号：Q575.11[生物学—生物化学] Q78