文献类型：期刊文献

英文题名：Complete genome sequence of the marine fish pathogen Vibrio anguillarum and genome-wide transposon mutagenesis analysis of genes essential for in vivo infection

作者：Yang Guanhua[1];Wang, Cheng[1];Wang, Xuetong[1];Ma, Ruiqing[1];Zheng, Huajun[4];Liu, Qin[1,3];Zhang, Yuanxing[1,3];Ma, Yue[1,3];Wang, Qiyao[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao, Peoples R China;[3]Shanghai Engn Res Ctr Maricultured Anim Vaccines, Shanghai, Peoples R China;[4]Chinese Natl Human Genome Ctr Shanghai, Shanghai MOST Key Lab Hlth & Dis Genom, Shanghai 201203, Peoples R China

年份：2018

卷号：216

起止页码：97

外文期刊名：MICROBIOLOGICAL RESEARCH

收录：;EI(收录号：20183605781856);WOS:【SCI-EXPANDED(收录号:WOS:000450244300011)】；

基金：This work was supported by grants from the Ministry of Agriculture of China (CARS-47-G17), the Science and Technology Commission of Shandong and Shanghai Municipality (2017CXGC0103 and 17391902000), and the Shanghai Pujiang Program (16PJD018). We thank Dr. Matthew K. Waldor from HHMI for his helpful comments and discussion on the manuscript.

语种：英文

外文关键词：Genome sequence; Tn-seq; Vibrio anguillarum; In vivo fitness; Conditionally essential genes

摘要：Vibrio anguillarum is a notorious bacterial pathogen that causes vibriosis in various marine farmed fish species. The highly pathogenic V. anguillarum strain MVM425 has been isolated from moribund turbot in the China Yellow Sea, but the genetic basis for its pathogenesis is undefined. Complete genome sequencing uncovered that the bacterium encodes 3985 protein-coding sequences (CDSs) on two chromosomes and 62 CDSs on the virulence plasmid pEIB1. To investigate the genes that are essential for its in vitro and in vivo growth, we applied transposon insertion sequencing technology (Tn-seq) to screen a highly saturated transposon insertion mutant library grown in Luria-Bertani broth with 2% NaCl (LB20) and in its natural host turbot. Tn-seq identified 473 and similar to 173 putative genes as essential for its growth in rich medium and the in vivo infection process, respectively. Additionally, our analysis revealed that the genes are associated to various fitness levels in the liver, spleen and kidney, suggesting different genetic requirements for colonization in these organs. Furthermore, we validated our Tn-seq data using gene knockout mutants and in vivo infection experiments. Comprehensive functional genomics analysis highlighted the conditionally essential genes for important pathways involved in energy metabolism, nitrogen metabolism, nucleotide synthesis, amino acid synthesis, cofactors, siderophore synthesis, secretion (T1SS, T2SS, and T6SS), quorum sensing, and flagellum biosynthesis. This study provides a wealth of information about V. anguillarum genes related to infection in fish and will facilitate the understanding of its pathogenesis as well as the development of diagnostics and vaccines against the pathogen.