Title: | Utilization of the ComRS system for the rapid markerless deletion of chromosomal genes in Streptococcus suis |
Author(s): | Zhu Y; Dong W; Ma J; Zhang Y; Pan Z; Yao H; |
Address: | "College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, PR China. Office International Des Epizooties (OIE) Reference Lab for Swine Streptococcosis, Nanjing Agricultural University, Nanjing 210095, PR China. Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, PR China" |
ISSN/ISBN: | 1746-0921 (Electronic) 1746-0913 (Linking) |
Abstract: | "AIM: To develop a markerless gene deletion strategy in Streptococcus suis to solve the problem that several serotypes against electrotransformation of foreign DNA. MATERIALS & METHODS: Bioinformatics retrieval was performed to identified ComRS systems functioning for natural transformation. A sacB-spc cassette with the upper and lower homologous fragments was amplification by fusion-PCR for spectinomycin-positive and sucrose-negative selection during gene deletion. RESULTS & CONCLUSION: Three phylogenetic clusters of ComR were identified to function for natural transformation by specific recognition to competence pheromone in S. suis. Thus, they were employed to establish gene deletion method. Its efficiency for genetic replacement was dependent on the length of homologs fragment and the concentration of donor DNA. This rapid gene-editing technique may greatly facilitate molecular studies on S. suis" |
Keywords: | "Amino Acid Sequence Animals Bacterial Proteins/*genetics Base Sequence Chromosomes, Bacterial/*genetics DNA Transformation Competence DNA, Bacterial/genetics Disease Models, Animal Female *Gene Deletion Gene Editing/*methods Gene Expression Regulation, Ba;" |
Notes: | "MedlineZhu, Yinchu Dong, Wenyang Ma, Jiale Zhang, Yue Pan, Zihao Yao, Huochun eng Research Support, Non-U.S. Gov't England 2019/01/22 Future Microbiol. 2019 Feb; 14:207-222. doi: 10.2217/fmb-2018-0279. Epub 2019 Jan 21" |