用于基因改造细菌基因组的策略：定点诱变、基因失活和基因过表达。
Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression.
作者信息
Xu Jian-zhong, Zhang Wei-guo
机构信息
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
出版信息
J Zhejiang Univ Sci B. 2016 Feb;17(2):83-99. doi: 10.1631/jzus.B1500187.
Abstract
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.
摘要
随着大肠杆菌或谷氨酸棒杆菌全基因组序列的可得，定向DNA操作策略迅速发展。DNA操作在理解基因功能以及根据需求构建新型工程菌方面发挥着重要作用。DNA操作包括修饰自体基因和表达异源基因。两种替代方法，即使用电穿孔线性DNA或重组自杀质粒，可实现多种DNA操作。然而，所需基因的过表达通常通过质粒介导来实现。本综述总结了用于对大肠杆菌和谷氨酸棒杆菌基因组进行基因改造的常见策略，并讨论了多层DNA操作的技术问题。提出了通过整合到基因组中来实现基因过表达的策略。本综述旨在为新手提供细菌基因组内DNA操作的易懂介绍，并为经验丰富的研究人员提供最新实验信息来源。
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