重组人肝细胞生长因子真核表达载体的构建
Construction of eukaryotic expression vector of recombinant human hepatocyte growth factor
宋旭飞 1董永红 2徐钧 3沈文涛 4周鹏4
作者信息
- 1. 030012 太原,山西省人民医院普外科;河南省安阳市人民医院
- 2. 030012 太原,山西省人民医院普外科
- 3. 山西医科大学第二医院普外科
- 4. 中国热带农业科学研究院生物技术研究所
- 折叠
摘要
目的 构建重组人肝细胞生长因子(rhHGF)的真核表达载体,为后续毕赤酵母蛋白高效表达提供基础.方法 根据酵母表达偏爱密码子合成rhHGF cDNA全长序列,将rhHGF和pGAPZαA质粒双酶切后行连接转化,构建穿梭载体pGAPZαA-rhHGF,并鉴定.结果 重组真核表达载体pGAPZA-rhHGF经限制性内切酶分析,与理论值相符,测序结果未见碱基变异.结论 成功构建了pGAPZαA-rhHGF真核表达载体,为后续毕赤酵母高效表达rhHGF蛋白提供了可行性.
Abstract
Objective To construct eukaryotic expression vector of recombinant human hepatocyte growth fac-tor (rhHGF), and to further provide the basis for the high expression of Pichia pastoris protein. Methods The rhHGF cDNA full-length sequence was synthesized according to the optimal codon of yeast expression. The inserted transfor-mation was performed after double digestion of rhHGF and pGAPZαA plasmids. The shuttle vector pGAPZαA-rhHGF was constructed and identified. Results The recombinant eukaryotic expression vector pGAPZαA-rhHGF was deter-mined by restriction endonuclease analysis and was consistent with the theoretical value. No mutated base sequence was found by sequencing analysis. Conclusion The eukaryotic expression vector pGAPZαA-rhHGF is successfully constructed, which further provids the feasibility for the high expression of rhHGF protein in Pichia pastoris.
关键词
肝细胞生长因子/酵母,巴斯德毕赤/表达Key words
Hepatocyte growth factor/Yeast/Pichia pastoris/Expression引用本文复制引用
基金项目
山西省科技攻关项目(20090311061-2)
出版年
2018
NETL
NSTL
维普
万方数据