胞内表达猪生长激素基因的重组酵母菌构建与鉴定  

Construction and identification of the intracellular expression recombinants for porcine growth hormone gene in Pichia pastoris

作　　者：付若彬[1] 刘海华[1] 张海英[1] 张莉[1] 刘世贵[1] 龙章富[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2007年第1期191-194,共4页Journal of Sichuan University(Natural Science Edition)

基　　金：教育部重点项目(国教00104);四川省"十五"重点项目(03NC002-001);四川省应用基础研究项目(02NY029-079)

摘　　要：将猪生长激素(pGH)cDNA定向插入毕赤酵母胞内表达载体pPIC 3.5k质粒,重组构建了pPIC 3.5k-pGH/GS115酵母表达系统,其表达产物的SDS-PAGE和Western blot结果表明,pGH蛋白在胞内获得正确表达,表达量达378 mg/L.The cDNA was oriently inserted for porcine growth hormone （pGH） into pPIC 3.5k, an expression vector for Pichia pastoris, which contains AOXI promoter and Pichia wild-type gene coding for histidinol dehydrogenase. The recombinant plasmid of pPIC 3.5k-pGH was linearnized by Sal Ⅰ and transformed into Pichia pastoris GS115 by eleetroporation. Five muilti-copy transformants were selected by PCR using their genome DNA samples as the templates. SDS-PAGE and Western blotting analysis showed that pGH gene expression produet bands appeared in intraeellular solution. The expressed pGH were aeeumulated up to 378 mg/L.

关 键 词：猪生长激素基因 毕赤酵母 真核表达 

分 类 号：Q81[生物学—生物工程]