| 重组融合蛋白rTMP-GH在巴斯德毕赤酵母中表达的研究 |
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| 引用本文： | 王崧,王军平,陈默,许杨,陈芳,申明强,王艾平,粟永萍,开丽.重组融合蛋白rTMP-GH在巴斯德毕赤酵母中表达的研究[J].第三军医大学学报,2008,30(21):2037-2040. |
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| 作者姓名： | 王崧 王军平 陈默 许杨 陈芳 申明强 王艾平 粟永萍 开丽 |
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| 作者单位： | 第三军医大学军事预防医学院防原医学教研室,全军复合伤研究所,创伤,烧伤与复合伤国家重点实验室,重庆,400038 |
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| 基金项目： | 国家高技术研究发展计划(863计划)，教育部全国优秀博士学位论文作者专项基金 |
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| 摘 要： | 目的 实现TPO模拟肽(thrombopoietin mimetic peptide,TMP)与人生长激素(human growth hormone,hGH)的重组融合蛋白rTMP-GH在巴氏毕赤酵母中的高效表达.方法 以含有rTMP-GH核苷酸序列的原核质粒为模板,PCR方法获得rTMP-GH的编码序列,并亚克降至载体pPIC9K中,将所构建pPIC9K-rTMP-GH表达质粒转化酵母宿主细胞GS115,MD/MM平板及G418抗性筛选高表达株,然后进行试管补料分批表达,Western blot对表达产物进行鉴定,通过巨核细胞集落形成能力对其,圭物学活性进行初步检测.结果 成功构建pPIC9K-rTMP-GH表达质粒,筛选到高表达rTMP-GH重组融合蛋白的巴氏毕赤酵母细胞株,初步证实所表达产物具有刺激巨核细胞集落形成的能力.结论 成功实现了rTMP-GH融合蛋白在巴氏毕赤酵母中的高效表达.
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| 关 键 词： | 毕赤酵母 表达 |
Expression and identification of recombinant fusion protein of thrombopoietin mimetic peptide and growth hormone in Pichia pastoris |
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| WANG Song,WANG Jun-ping,CHEN Mo,XU Yang,CHEN Fang,SHEN Ming-qiang,WANG Ai-ping,SU Yong-ping,KAI Li.Expression and identification of recombinant fusion protein of thrombopoietin mimetic peptide and growth hormone in Pichia pastoris[J].Acta Academiae Medicinae Militaris Tertiae,2008,30(21):2037-2040. |
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| Authors: | WANG Song WANG Jun-ping CHEN Mo XU Yang CHEN Fang SHEN Ming-qiang WANG Ai-ping SU Yong-ping KAI Li |
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| Institution: | WANG Song,WANG Jun-ping,CHEN Mo,XU Yang,CHEN Fang,SHEN Ming-qiang,WANG Ai-ping,SU Yong-ping,KAI Li(State Key Laboratory of Trauma,Burns , Combined Injury,Institute of Combined Injury,College of Preventive Medicine,Third Military Medical University,Chongqing 400038,China) |
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| Abstract: | Objective To fulfill high-efficient expression of rTMP-GH,a recombinant fusion protein of thrombopoietin mimetic peptide and human growth hormone,in Pichia pastoris.Methods cDNA fragment coding for rTMP-GH was amplified by PCR,and subsequently cloned into the expression vector pPIC9K.The linearlized plasmid pPIC9K-rTMP-GH was transformed into Pichia pastoris GS115 cells and screened in MD/MM plates under pressure of G418.The recombinant fusion protein rTMP-GH was identified by Western blotting,and its biolo... |
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| Keywords: | rTMP-GH |
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