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基于转座子的基因递送系统在细菌中的应用。

Applications of transposon-based gene delivery system in bacteria.

作者信息

Choi Kyoung-Hee, Kim Kang-Ju

机构信息

Department of Oral Microbiology, College of Dentistry, Wonkwang University, Iksan 570-749, Korea.

出版信息

J Microbiol Biotechnol. 2009 Mar;19(3):217-28. doi: 10.4014/jmb.0811.669.

Abstract

Mobile genetic segments, or transposons, are also referred to as "jumping genes" as they can shift from one position in the genome to another, thus inducing a chromosomal mutation. According to the target site-specificity of the transposon during a transposition event, the result is either the insertion of a gene of interest at a specific chromosomal site, or the creation of knockout mutants. The former situation includes the integration of conjugative transposons via site-specific recombination, several transposons preferring a target site of a conserved ATrich sequence, and Tn7 being site-specifically inserted at attTn7, the downstream of the essential glmS gene. The latter situation is exploited for random mutagenesis in many prokaryotes, including IS (insertion sequence) elements, mariner, Mu, Tn3 derivatives (Tn4430 and Tn917), Tn5, modified Tn7, Tn10, Tn552, and Ty1, enabling a variety of genetic manipulations. Randomly inserted transposons have been previously employed for a variety of applications such as fmgenetic footprinting, gene transcriptional and translational fusion, signature-tagged mutagenesis(STM),DNA or cDNA sequencing, transposon site hybridization(TraSH), and scanning linker mutagenesis(SLM). Therefore, transposon-mediated genetic engineering is a baluable discipiline for the study of bacterial physiology and pathogenesis in living hosts.

摘要

移动遗传片段,即转座子,也被称为“跳跃基因”,因为它们能够从基因组中的一个位置转移到另一个位置,从而诱发染色体突变。根据转座过程中转座子的靶位点特异性,结果要么是将感兴趣的基因插入到特定的染色体位点,要么是产生基因敲除突变体。前一种情况包括通过位点特异性重组整合接合转座子,几种转座子倾向于保守的富含A序列的靶位点,以及Tn7特异性插入到必需的glmS基因下游的attTn7位点。后一种情况被用于许多原核生物的随机诱变,包括IS(插入序列)元件、水手转座子、Mu、Tn3衍生物(Tn4430和Tn917)、Tn5、修饰的Tn7、Tn10、Tn552和Ty1,从而实现多种遗传操作。随机插入的转座子先前已被用于多种应用,如遗传足迹分析、基因转录和翻译融合、签名标签诱变(STM)、DNA或cDNA测序、转座子位点杂交(TraSH)以及扫描接头诱变(SLM)。因此,转座子介导的基因工程是研究活宿主中细菌生理学和发病机制的宝贵学科。

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