Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression

作     者：Jian-zhong XU Wei-guo ZHANG 

作者机构：The Key Laboratory of Industrial Biotechnology Ministry of Education School of Biotechnology Jiangnan University 

出 版 物：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 (浙江大学学报（英文版）B辑（生物医学与生物技术）)

年 卷 期：2016年第17卷第2期

页      面：83-99页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学] 

基　　金：supported by the Natural Science Foundation of Jiangsu Province(No.BK20150149) the Fundamental Research Funds for the Central Universities(No.JUSRP51504) the Youth Foundation of Jiangnan University(No.JUSRP115A19),China 

主　　题：Escherichia coli Corynebacterium glutamicum DNA manipulation Site-directed mutagenesis Gene inactivation Gene over-expression 

摘      要：With the availability of the whole genome sequence of Escherichia coli or Corynebacterium glutamicum, strategies for directed DNA manipulation have developed rapidly. DNA manipulation plays an important role in understanding the function of genes and in constructing novel engineering bacteria according to requirement. DNA manipulation involves modifying the autologous genes and expressing the heterogenous genes. Two alternative approaches, using electroporation linear DNA or recombinant suicide plasmid, allow a wide variety of DNA manipulation. However, the over-expression of the desired gene is generally executed via plasmid-mediation. The current review summarizes the common strategies used for genetically modifying E. coli and C. glutamicum genomes, and discusses the technical problem of multi-layered DNA manipulation. Strategies for gene over-expression via integrating into genome are proposed. This review is intended to be an accessible introduction to DNA manipulation within the bacterial genome for novices and a source of the latest experimental information for experienced investigators.