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题名: Gene Deletion Strategy To Examine the Involvement of the Two Chondroitin Lyases in Flavobacterium columnare Virulence
作者: Li, Nan1; Qin, Ting1; Zhang, Xiao Lin1; Huang, Bei2; Liu, Zhi Xin1; Xie, Hai Xia1; Zhang, Jin1; McBride, Mark J.3; Nie, Pin1
刊名: APPLIED AND ENVIRONMENTAL MICROBIOLOGY
发表日期: 2015-11-01
DOI: 10.1128/AEM.01586-15
卷: 81, 期:21, 页:7394-7402
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Biotechnology & Applied Microbiology ; Microbiology
研究领域[WOS]: Biotechnology & Applied Microbiology ; Microbiology
英文摘要: Flavobacterium columnare is an important bacterial pathogen of freshwater fish that causes high mortality of infected fish and heavy economic losses in aquaculture. The pathogenesis of this bacterium is poorly understood, in part due to the lack of efficient methods for genetic manipulation. In this study, a gene deletion strategy was developed and used to determine the relationship between the production of chondroitin lyases and virulence. The F. johnsoniae ompA promoter (PompA) was fused to sacB to construct a counterselectable marker for F. columnare. F. columnare carrying PompA-sacB failed to grow on media containing 10% sucrose. A suicide vector carrying PompA-sacB was constructed, and a gene deletion strategy was developed. Using this approach, the chondroitin lyase-encoding genes, cslA and cslB, were deleted. The Delta cslA and Delta cslB mutants were both partially deficient in digestion of chondroitin sulfate A, whereas a double mutant (Delta cslA Delta cslB) was completely deficient in chondroitin lyase activity. Cells of F. columnare wild-type strain G(4) and of the chondroitin lyase-deficient Delta cslA Delta cslB mutant exhibited similar levels of virulence toward grass carp in single-strain infections. Coinfections, however, revealed a competitive advantage for the wild type over the chondroitin lyase mutant. The results indicate that chondroitin lyases are not essential virulence factors of F. columnare but may contribute to the ability of the pathogen to compete and cause disease in natural infections. The gene deletion method developed in this study may be employed to investigate the virulence factors of this bacterium and may have wide application in many other members of the phylum Bacteroidetes.
关键词[WOS]: AC LYASE ; GLIDING MOTILITY ; CYTOPHAGA JOHNSONAE ; FISH ; G(4) ; CONSTRUCTION ; MUTAGENESIS ; INSERTIONS ; CLONING ; GENOME
语种: 英语
项目资助者: National Basic Research Program of China (973 Program)(2009CB118703) ; State Key Laboratory of Freshwater Ecology and Biotechnology ; National Natural Science Foundation of China(31001135)
WOS记录号: WOS:000363462900006
ISSN号: 0099-2240
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内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/27340
Appears in Collections:鱼类生物学及渔业生物技术研究中心_期刊论文

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作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Hubei Province, Peoples R China
2.Jimei Univ, Coll Fisheries, Xiamen, Fujian Province, Peoples R China
3.Univ Wisconsin, Dept Biol Sci, Milwaukee, WI 53201 USA
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