| 双表达盒人骨唾液酸蛋白毕赤酵母工程细胞的构建 |
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| 引用本文： | 王征,王捷,杨静,张宏斌,武婕,魏春华.双表达盒人骨唾液酸蛋白毕赤酵母工程细胞的构建[J].中国生化药物杂志,2005,26(5):264-266,269. |
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| 作者姓名： | 王征 王捷 杨静 张宏斌 武婕 魏春华 |
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| 作者单位： | 广州军区广州总医院,医学实验中心,广东,广州,510010 |
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| 基金项目： | 广东省自然科学基金研究团队项目（No.20023001） |
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| 摘 要： | 目的构建含人骨唾液酸蛋白(hBSP)双表达盒的毕赤酵母工程细胞,为hBSP在毕赤酵母中的高效非融合分泌表达奠定基础。方法用PCR方法扩增hBSP基因,将其亚克隆到毕赤酵母表达载体pPICZαA中,并在此基础上构建含有双表达盒的重组pPICZαAN-2×hBSP载体,电转化毕赤酵母GS115,通过Zeocin高抗性筛选转化子并对其表型进行PCR鉴定。结果得到GS115/pPICZαAN-hBSP和GS115/pPICZαAN-2×hBSP毕赤酵母工程细胞。结论GS115/pPICZαAN-2×hBSP毕赤酵母工程细胞中含双表达盒hBSP。
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| 关 键 词： | 人骨唾液酸蛋白 毕赤酵母 表达盒 |
| 文章编号： | 1005-1678（2005）05-0264-03 |
| 收稿时间： | 2004-11-22 |
| 修稿时间： | 2004-11-22 |
Construction of recombinant Pichia pastoris containing two expression cassettes of human bone sialoprotein |
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| WANG Zheng,WANG Jie,YANG Jing,ZHANG Hong-bin,WU Jie,WEI Chun-hua.Construction of recombinant Pichia pastoris containing two expression cassettes of human bone sialoprotein[J].Chinese Journal of Biochemical Pharmaceutics,2005,26(5):264-266,269. |
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| Authors: | WANG Zheng WANG Jie YANG Jing ZHANG Hong-bin WU Jie WEI Chun-hua |
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| Abstract: | PurposeTo construct recombinant Pichia pastoris containing two expression cassettes of rhuman bone sialoprotein(rhBSP) and to establish the basis for high expression of secreted no-fusing rhBSP.MethodsHuman BSP gene was amplified by PCR and subcloned into Pichia pastoris expression vector pPICZaA.On the basis of pPICZaAN-hBSP,the recombinant vector pPICZαAN-2×hBSP containing two expression cassettes was constructed and transformed to Pichia pastoris GS115 by electroporation.The positive transformants were screened by increasing Zeocin concentration and identified by PCR.ResultsThe recombinant Pichia pastoris GS115/pPICZαAN-hBSP and GS115/pPICZαAN-2×hBSP were obtained.ConclusionGS115/pPICZαAN-2×hBSP contains two expression cassettes of rhBSP. |
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| Keywords: | human bone sialoprotein Pichia pastoris expression cassette |
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