Uehara, Tsuyoshi and Prathapam, Ramadevi (2018) A temperature-sensitive replicon enables efficient gene inactivation in Pseudomonas aeruginosa. Journal of microbiological methods, 144. pp. 47-52. ISSN 01677012

Abstract

Tools to enable genome editing are essential for understanding physiology. Here we report a gene replacement method in Pseudomonas aeruginosa using a temperature-sensitive replicon plasmid that does not require mating or isolation of a merodiploid intermediate. This approach was validated by replacing the non-essential ampD gene with a gentamicin resistance cassette. In addition lpxA and lpxD, both located in a complex gene cluster including multiple downstream essential genes, were inactivated when complemented by each target gene in trans. These strains did not grow when expression of the gene in trans was repressed, confirming that both genes are essential for viability. This method facilitates efficient gene inactivation in P. aeruginosa.

Item Type:	Article
Keywords:	Gene inactivation Pseudomonas aeruginosa Temperature-sensitive replicon
Date Deposited:	09 Dec 2017 00:45
Last Modified:	25 Jan 2019 00:45
URI:	https://oak.novartis.com/id/eprint/34295