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

被引：14
作者：
Xu, Jian-zhong [1 ]
Zhang, Wei-guo [1 ]
机构：
[1] Jiangnan Univ, Sch Biotechnol, Minist Educ, Key Lab Ind Biotechnol, Wuxi 214122, Peoples R China
来源：
JOURNAL OF ZHEJIANG UNIVERSITY-SCIENCE B | 2016年 / 17卷 / 02期
关键词：
Escherichia coli; Corynebacterium glutamicum; DNA manipulation; Site-directed mutagenesis; Gene inactivation; Gene over-expression; POLYMERASE-CHAIN-REACTION; DOUBLE-STRANDED DNA; L-LYSINE PRODUCTION; PCR-BASED METHOD; CORYNEBACTERIUM-GLUTAMICUM; ESCHERICHIA-COLI; HOMOLOGOUS RECOMBINATION; POSITIVE-SELECTION; BACILLUS-SUBTILIS; EFFICIENT METHOD;
D O I：
10.1631/jzus.B1500187
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
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.
引用
收藏
页码：83 / 99
页数：17
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