| 人骨唾液酸蛋白酵母工程细胞的构建与表达 |
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| 引用本文： | 董燕,王捷,张宏斌,武捷,郑文岭.人骨唾液酸蛋白酵母工程细胞的构建与表达[J].中国生化药物杂志,2003,24(3):109-112. |
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| 作者姓名： | 董燕 王捷 张宏斌 武捷 郑文岭 |
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| 作者单位： | 广州军区广州总医院,医学实验科,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 (2 0 0 2 3 0 0 1) |
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| 摘 要： | 目的构建表达重组人骨唾液酸蛋白 (rhBSP)的巴斯德毕赤酵母工程细胞 ,实现rhBSP的酵母胞外分泌性高表达。方法通过PCR扩增获得人BSP基因后 ,构建重组表达载体pPICZaA hbsp ,经电转化导入到毕赤酵母GS115菌中 ,获得GS115 pPICZaA hbsp工程菌 ,通过Zeocin高抗性筛选和摇瓶发酵筛选出高表达rhBSP的菌株 ,并进行高密度发酵提高表达量。结果rhBSP分子量接近 6 6 0 0 0 ,产物可达上清液总蛋白的 80 % ,浓度约 0 .2 4 8g L。结论基因工程菌GS115 pPICZaA hbsp能高效地表达rhBSP并分泌到胞外
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| 关 键 词： | 人骨唾液酸蛋白 毕赤酵母 基因表达 发酵 |
| 文章编号： | 1005-1678(2003)03-0109-04 |
| 修稿时间： | 2002年7月26日 |
Construction of recombinant Pichia pastoris expressing recombinant human bone sialoprotein |
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| DONG Yan,WANG Jie,ZHANG Hong bin,WU Jie,ZHENG Wen ling.Construction of recombinant Pichia pastoris expressing recombinant human bone sialoprotein[J].Chinese Journal of Biochemical Pharmaceutics,2003,24(3):109-112. |
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| Authors: | DONG Yan WANG Jie ZHANG Hong bin WU Jie ZHENG Wen ling |
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| Abstract: | PurposeTo construct recombinant Pichia pastoris strain which can express and secrete recombinant human bone sialoprotein(rhBSP) highly.MethodsGene of rhBSP amplified by PCR was recombined with expressive vector pPICZaA. Recombinant vector was transformed to Pichia pastoris GS115 by electroporation. Strains GS115/pPICZaA hbsp highly expressing hBSP were screened by increasing Zeocin concentration and fermentation in flasks. The expressing level was increased by high density fermentation.ResultsrhBSP is approximate 66 000 with specific antigenicity,whose concentration is about 0.248 g/L, 80% of total protein in supernatant.ConclusionRecombinant strains GS115/pPICZaA hbsp can highly express and secrete rhBSP. |
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| Keywords: | human bone sialoprotein Pichia pastoris gene expression fermentation |
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