High-Efficiency, Two-Step Scarless-Markerless Genome Genetic Modification in Salmonella enterica

Current Microbiology
Shizhong GengXinan Jiao

Abstract

We present a two-step method for scarless-markerless genome genetic modification in Salmonella enterica based on the improved suicide plasmid pGMB152. The whole LacZYA gene can provide a lacZ-based blue/white screening strategy for fast selection of double-crossover mutants by allelic exchange. The high efficiency of this genetic engineering strategy permits the study of gene function by gene knockin, site-directed mutagenesis, and gene knockout to construct live attenuated vaccines.

References

Jul 1, 1996·Molecular Microbiology·J McFadden
Aug 26, 1998·Infection and Immunity·J M ReyratR Rappuoli
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Feb 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·Wanyin DengB Brett Finlay
Apr 28, 2004·Plasmid·Nadège PhilippeDominique Schneider
Aug 13, 2009·Journal of Biomedicine & Biotechnology·Shi-Zhong GengXiang Chen
Oct 2, 2012·Journal of Microbiological Methods·Luitpold FriedKirsten Jung

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Methods Mentioned

BETA
genetic modifications
gene knockout
genetic modification
PCR
Gene
gene knockin

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