Knockout of theadpgene related with colonization inBacillus nematocida B16 using customized transcription activator-like effectors nucleases

Qiuhong Niu, Haoying Zheng, Lin Zhang, Fujun Qin, Loryn Facemire, Guo Zhang, Feng Cao, Ke-qin Zhang, Xiaowei Huang, Jianwei Yang, Lei He, Chanjuan Liu
2015 Microbial Biotechnology  
Bacillus nematocida B16 is able to dominate in the intestines of the worm Caenorhabditis elegans in 'Trojan horse' pathogenic mechanism. The adp is one candidate gene which potentially play a vital role in the colonization from our previous random mutagenesis screening results. To analyse the functional role of this gene, we constructed the adp knockout mutant through customized transcription activator-like effectors nucleases (TALEN), which has been successfully used in yeasts, nematodes,
more » ... fish and human pluripotent cells. Here, we first time report this knockout method in bacteria on this paper. Bioassay experiments demonstrated that the adp knockout mutant of B16 showed considerably lower colonization activity, reduced numbers of intestines and less than 80% nematocidal activity compared with the wild-type strain when infected for 48 h. However, no obvious change on proteolytic activity was observed in the mutant. Conversely, the complementation of adp gene restored most of the above deficient phenotypes. These results indicated that the adp gene was involved in surface adhesion and played a comparatively important role in colonizing host nematodes. Moreover, TALENs successfully disrupt target genes in bacteria.
doi:10.1111/1751-7915.12282 pmid:25912819 pmcid:PMC4476823 fatcat:3n6whs62yjcx5duh4mzf3nhueu