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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
【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.
【Abstract】 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 SalⅠand transformed into Pichia pastoris GS115 by electroporation.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 product bands appeared in intracellular solution.The expressed pGH were accumulated up to 378 mg/L.
- 【文献出处】 四川大学学报(自然科学版) ,Journal of Sichuan University(Natural Science Edition) , 编辑部邮箱 ,2007年01期
- 【分类号】Q78
- 【被引频次】3
- 【下载频次】200