Prophage recombinases-mediated genome engineering in Lactobacillus plantarum

Microbial Cell Factories

Research

Qingsheng Qi¹  Jing Wang¹  Peng Yang¹ 

[1] State Key Laboratory of Microbial Technology, Shandong University, 250100, Jinan, People’s Republic of China;

关键词: Homologous recombination;    Lactic acid bacteria;    Prophage;    Genome modification;    Marker-free;   

DOI  :  10.1186/s12934-015-0344-z

 received in 2015-07-07, accepted in 2015-09-18,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundLactobacillus plantarum is a food-grade microorganism with industrial and medical relevance belonging to the group of lactic acid bacteria (LAB). Traditional strategies for obtaining gene deletion variants in this organism are mainly vector-based double-crossover methods, which are inefficient and laborious. A feasible possibility to solve this problem is the recombineering, which greatly expands the possibilities for engineering DNA molecules in vivo in various organisms.ResultsIn this work, a double-stranded DNA (dsDNA) recombineering system was established in L. plantarum. An exonuclease encoded by lp_0642 and a potential host-nuclease inhibitor encoded by lp_0640 involved in dsDNA recombination were identified from a prophage P1 locus in L. plantarum WCFS1. These two proteins, combined with the previously characterized single strand annealing protein encoded by lp_0641, can perform homologous recombination between a heterologous dsDNA substrate and host genomic DNA. Based on this, we developed a method for marker-free genetic manipulation of the chromosome in L. plantarum.ConclusionsThis Lp_0640-41-42-mediated recombination allowed easy screening of mutants and could serve as an alternative to other genetic manipulation methods. We expect that this method can help for understanding the probiotic functionality and physiology of LAB.

【 授权许可】

CC BY   
© Yang et al. 2015