摘要
为大量获得具有生物活性的大黄鱼生长激素(Pseucdosciaena crocea,Growth Hormone,pcGH),利用本实验室分离的天然大黄鱼生长激素基因在毕赤酵母表达系统中的表达进行了研究。把pcGH克隆到酵母整合型质粒载体中构建重组表达载体pPIC9K-pcGH,电击转化将pcGH转化进his4缺陷型毕赤酵母GSll5菌株染色体上,经甲醇诱导得以高效表达,SDS-PAGE证实了表达产物为大黄鱼生长激素,表达量约为62.5mg/L。
This study aimed at high-yield expression of the pcGHgene which was separated from natural large yellow croaker in the yeast Piehia pastoris in order to produce amount of large yellow croaker growth hormone （Pseucdosciaena crocea, Growth Hormone, pcGH） with the feature of biological activity for further use. The pcGHgene was first cloned into the integrative expression vector of methylotrophic yeast P. Pastoris to form the recombinant vector pPIC9K-pcGH, and the linearized recombinant plasmid was then transformed into the chromosome of the HIS4 mutant yeast GS115 by electroporation, and it could finally get high-yield expression induced by methanol. SDS-PAGE confirmed that the expression product was pcGH, and it led to a level as high as of 62.5 mg/L.
出处
《中国农学通报》
CSCD
2013年第17期70-74,共5页
Chinese Agricultural Science Bulletin
基金
福建省科技术项目"大黄鱼基因工程促生长饲料的研制"(98-I-11)
