Advanced genetic strategies for recombinant protein expression in Escherichia coli

被引:665
作者
Sorensen, HP [1 ]
Mortensen, KK [1 ]
机构
[1] Aarhus Univ, Dept Mol Biol, Lab BioDesign, DK-8000 Aarhus C, Denmark
关键词
Escherichia coli; recombinant protein expression systems; inclusion bodies; fusion proteins; rare codon tRNAs;
D O I
10.1016/j.jbiotec.2004.08.004
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Preparations enriched by a specific protein are rarely easily obtained from natural host cells. Hence, recombinant protein production is frequently the sole applicable procedure. The ribosomal machinery, located in the cytoplasm is an outstanding catalyst of recombinant protein biosynthesis. Escherichia coli facilitates protein expression by its relative simplicity, its inexpensive and fast high-density cultivation, the well-known genetics and the large number of compatible tools available for biotechnology. Especially the variety of available plasmids, recombinant fusion partners and mutant strains have advanced the possibilities with E. coli. Although often simple for soluble proteins, major obstacles are encountered in the expression of many heterologous proteins and proteins lacking relevant interaction partners in the E. coli cytoplasm. Here we review the current most important strategies for recombinant expression in E. coli. Issues addressed include expression systems in general, selection of host strain, mRNA stability, codon bias, inclusion body formation and prevention, fusion protein technology and site-specific proteolysis, compartment directed secretion and finally co-overexpression technology. The macromolecular background for a variety of obstacles and genetic state-of-the-art solutions are presented. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:113 / 128
页数:16
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