摘要
外源蛋白在表达宿主中的可溶性是影响其应用的关键因素。采用m Cherry报告基因,构建Msi R-m Cherry融合蛋白,通过测定荧光值定性定量的测定Msi R蛋白在大肠杆菌中的可溶性表达量。随后,利用易错聚合酶链反应(Errorprone-PCR)和致突变菌株(XL1-Red)两种方式构建了msi R-m Cherry基因突变文库,经筛选得到了4株在大肠杆菌中以37℃作为诱导温度下可溶性明显提高的蛋白突变体。
The soluble expression of heterologous protein in expression host is a key factor affecting their applications, mCherry, a fluorophore reporter gene, was used as a fusion protein to analyze the soluble expression of MsiR protein quantitatively in E. coli. Errorprone PCR and XL1-Red were applied in constructing a msiR-mCherty gene mutant library. Four mutant strains improving the soluble expression of MsiR protein at 37℃ in E. coli were finally obtained.
出处
《中国酿造》
CAS
北大核心
2015年第5期43-47,共5页
China Brewing
基金
天津市科技计划项目(13ZCZDSY05100)
