大肠杆菌中低分子量重组蛋白产量的增加。
Increased production of low molecular weight recombinant proteins in Escherichia coli.
作者信息
Belagaje R M, Reams S G, Ly S C, Prouty W F
机构信息
Department of Biotechnology, Eli Lilly and Company, Indianapolis, Indiana 46285, USA.
出版信息
Protein Sci. 1997 Sep;6(9):1953-62. doi: 10.1002/pro.5560060916.
Abstract
A general method for obtaining high-level production of low molecular weight proteins in Escherichia coli is described. This method is based on the use of a novel Met-Xaa-protein construction which is formed by insertion of a single amino acid residue (preferably Arginine or Lysine) between the N-terminal methionine and the protein of interest. The utility of this method is illustrated by examples for achieving high-level production of human insulin-like growth factor-1, human proinsulin, and their analogs. Furthermore, highly produced insulin-like growth factor-1 derivatives and human proinsulin analogs are converted to their natural sequences by removal of dipeptides with cathepsin C.
摘要
描述了一种在大肠杆菌中获得低分子量蛋白质高水平表达的通用方法。该方法基于使用一种新型的甲硫氨酸-Xaa-蛋白质构建体，它是通过在N端甲硫氨酸和目标蛋白质之间插入单个氨基酸残基（优选精氨酸或赖氨酸）形成的。通过实现人胰岛素样生长因子-1、人胰岛素原及其类似物的高水平表达的实例说明了该方法的实用性。此外，通过组织蛋白酶C去除二肽，将高表达的胰岛素样生长因子-1衍生物和人胰岛素原类似物转化为它们的天然序列。
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