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

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• 作者：Jian-zhong Xu ; Wei-guo Zhang
• 关键词：Escherichia coli ; Corynebacterium glutamicum ; DNA manipulation ; Site ; directed mutagenesis ; Gene inactivation ; Gene over ; expression ; 大肠杆菌 ; 谷氨酸棒杆菌 ; DNA 操作方法 ; 定点突变 ; 基因失活 ; 基因过表达
Q754
• 刊名：Journal of Zhejiang University SCIENCE B
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：17
• 期：2
• 页码：83-99
• 全文大小：960 KB
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• 作者单位：Jian-zhong Xu (1)
  Wei-guo Zhang (1)
  
  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China
  
• 刊物主题：Biomedicine general;
• 出版者：Springer Berlin Heidelberg
• ISSN：1862-1783

文摘

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. Keywords Escherichia coli Corynebacterium glutamicum DNA manipulation Site-directed mutagenesis Gene inactivation Gene over-expression