Rapid method of luxS and pfs gene inactivation in enterotoxigenic Escherichia coli and the effect on biofilm formation

• Published in: Molecular medicine reports Vol. 13; no. 1; pp. 257 - 264
• Main Authors: WANG, XIAOXU, LI, SHIYU, LU, XIAORAN, HU, PAN, CHEN, HAIYAN, LI, ZHENG, BU, ZHAOYANG, LANG, XULONG, WANG, XINGLONG
• Format: Journal Article
• Language: English
• Published: Greece D.A. Spandidos 01-01-2016; Spandidos Publications; Spandidos Publications UK Ltd
• Subjects: Antibiotic resistance; auto-inducer 2; Bacteria; Bacterial infections; Bioengineering; biofilm formation; Biofilms; Biotechnology; Chromosomes; Chromosomes, Bacterial - genetics; Cloning; Deoxyribonucleic acid; Diarrhea; DNA; Drug resistance; E coli; Electrophoresis, Agar Gel; Enterotoxigenic Escherichia coli - genetics; Enterotoxins; Enzymes; Escherichia coli; Gene expression; gene inactivation; Gene Silencing; Genes, Bacterial; Genetic engineering; Genetic Markers; Genetic Techniques; Genetic Vectors - metabolism; Homoserine - analogs & derivatives; Homoserine - metabolism; Lactones - metabolism; LuxS protein; Metabolic engineering; Microbial mats; Microorganisms; Mutation - genetics; Plasmids; Polymerase Chain Reaction; Population density; quorum sensing; Recombination; Recombination, Genetic - genetics; Sequence Analysis, DNA; Strains (organisms); Studies; China
• ISSN: 1791-2997; 1791-3004
• DOI: 10.3892/mmr.2015.4532

Rapid and efficient inactivation of a target gene in Escherichia coli chromosomes is required to investigate metabolic engineering. In the present study, a multiple gene inactivation approach was demonstrated in four strains of enterotoxigenic E. coli (ETEC), which are the predominant pathogenic bacteria causing piglet diarrhea, mediated by λ Red and Xer recombination. The chromosomal genes, luxS and pfs were inactivated using the multiple gene inactivation approach in the wild-type strains of E. coli, K88, K99, 987P and F41. This indicated that dif sites may be reused to inactivate multiple chromosomal genes when no antibiotic-resistant selectable markers remain. Following inactivation of luxS and pfs, the ability of ETEC to produce the quorum sensing signal, and induce auto-inducer 2 activity and biofilm formation were significantly reduced. Furthermore, the multiple gene inactivation approach also exhibits a high recombination efficiency and follows a simple process.