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.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
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 )&&
关键词
鲤鱼生长激素
毕赤酵母
胞内表达
促生长作用
Cyprinus carpio growth hormone, P. pastoris, intracellular expression, growth promoting
