文献类型：期刊文献

中文题名：溶藻弧菌dldh基因突变株的构建及其生物学功能研究

英文题名：Construction and Characterization of a Mutant of Vibrio alginolyticus ZJ03 Δdldh Strain

作者：庞欢瑛[1,2,3];陈立明[1,2,3];黄郁葱[1,2,3];简纪常[1,2,3];鲁义善[1,2,3];吴灶和[2,3,4]

机构：[1]广东海洋大学水产学院;[2]广东省水产经济动物病原生物学及流行病学重点实验室;[3]广东省教育厅水产经济动物病害控制重点实验室;[4]仲恺农业工程学院

年份：2014

卷号：0

期号：10

起止页码：161

中文期刊名：生物技术通报

外文期刊名：Biotechnology Bulletin

收录：CSTPCD、、北大核心2011、CSCD_E2013_2014、北大核心、CSCD

基金：国家科技支撑计划(2012BAD17B02;2012 BAD17B03);广东省自然科学基金项目(S201304 0014562)

语种：中文

中文关键词：溶藻弧菌;dldh缺失株;生物特性

外文关键词：Vibrio alginolyticus dldh mutant Biological characteristics

中文摘要：为揭示二氢硫辛酰胺脱氢酶(Dihydrolipoamide dehydrogenase,DLDH)在溶藻弧菌(Vibrio alginolyticus)致病中的作用,以pRE112自杀质粒为载体,应用插入失活突变法构建dldh基因插入突变株,比较分析了野生株ZJ03与缺失株ZJ03Δdldh在生长、泳动能力、酶活性、生物膜形成、致病性的表现。结果表明,dldh基因的突变,使溶藻弧菌生长的迟缓期延长、泳动能力显著减弱(P<0.05),酶活性显著降低(P<0.05),生物膜生成显著减少(P<0.05),对石斑鱼(Epinephelus coioides)的致病性也明显减弱(P<0.05)。本研究为从细菌能量代谢角度研究弧菌的致病机制提供理论依据。

外文摘要：Vibrio alginolyticus, a gram-negative bacterium has been described to be among the most common and economically important aquatic pathogen of fish and shellfish. Dihydrolipoamide dehydrogenase （ DLDH ） is homodimeric flavoproteins that catalyse the NAD1 dependent reoxidation of dihydrolipoamide in a number of muhienzyme complexes. This enzyme classically involved in energy metabolism. To identify DLDH＇ s role during infection, dldh gene deletion strain （ ZJO3Adldh ） were constructed using the insertion mutation method. With the pRE112 suicide plasmid as the carrier, the Overlap PCR and homologous recombination technology were used to construct the mutant strains. Furthermore, the changes of the physiology and pathogenicity of ZJO3Adldh mutant strains compared with the strain ZJ03, such as growth, swarming ability, biofilm and LD50 were investigated. The growth curve showed that the mutant grew slowly in lag phase compared to the wild- type strain, and the swarming ability, enzyme activity and biofilm formation of ZJO3Adldh mutant strains showed significant reduced （ P〈0.05 ） . The fish lethal test showed that the virulence of the ZJO3Adldh mutant strains was 102 times lower than the ZJ03 strain. In conclusion, the ZJO3Adldh mutant strains were constructed successfully, and this study showed that enzymes classically involved in energy metabolism may play an important role in pathogenic mechanism of V. alginolyticus.