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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

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【作者】 付若彬刘海华张海英张莉刘世贵龙章富

【Author】 FU Ruo-bin1,LIU Hai-hua1,ZHANG Hai-ying1,ZHANG Li1,LIU Shi-gui1,2,LONG Zhang-fu1,2(1.Key Laboratory of Bio-Resources and Eco-Environment Ministry of Education,College of Life Sciences,Sichuan UniversityChengdu 610064,China;2.Institute of Chengdu Chuangxin Biotechnology,Sichuan University,Chengdu 610064,China)

【机构】 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成都610064成都川大创新生物技术研究所

【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.

【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)
  • 【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
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