迟缓爱德华氏菌中甘油醛-3-磷酸脱氢酶胞外分泌调控.PDF

微生物学通报 Oct. 20, 2017, 44(10): 2398−2406 Microbiology China /wswxtbcn tongbao@ DOI: 10.13344/j.microbiol.china.170271 研究报告 迟缓爱德华氏菌中甘油醛-3-磷酸脱氢酶的胞外分泌调控 2 1,2* 邓佳 吴海珍 (1. 200237) (2. 200237) 摘 要：【目的】迟缓爱德华氏菌甘油醛-3-磷酸脱氢酶(GAPDH)是糖酵解途径中关键酶之一， 前期研究证实是一种广谱性抗原，可作为水产养殖细菌病免疫防治中疫苗的开发靶点。本文探 究迟缓爱德华氏菌甘油醛-3-磷酸脱氢酶的胞外分泌机制。【方法】通过 Western blot 和ELISA 方法考察迟缓爱德华氏菌经典分泌系统缺失株GAPDH 胞外分泌情况；使用ELISA 方法对迟缓 爱德华氏菌突变体文库的GAPDH 胞外分泌进行了大规模筛查，并结合qRT-PCR 对筛查得到的 插入失活株进行了表达分析。【结果】经典分泌系统与 GAPDH 的胞外分泌存在一定相关性。 突变体文库的大规模筛查得到两株GAPDH 分泌量明显增加的插入失活株ΔesrA 和ΔesrC ，这 两个基因的失活会导致GAPDH 的胞外分泌量显著上调。【结论】迟缓爱德华氏菌GAPDH 的 胞外分泌受EsrA 和EsrC 负调控。 关键词：迟缓爱德华氏菌，GAPDH ，胞外分泌，突变体文库 Regulation of GAPDH secretion in Edwardsiella tarda DENG Jia2 WU Hai-Zhen 1,2* (1. State Key Laboratory of Bioreactor Engineering, Shanghai 200237, China) (2. East China University of Science and Technology School of Biotechnology, Shanghai 200237, China) Abstract: [Objective] As one of the key enzymes of the glycolysis in E . tarda, GAPDH was previously identified to be a broad-spectrum antigen and could be used as the target of bacterial vaccine design in aquaculture. We explore the mechanism of secretion of GAPDH in E . tarda. [Methods] Secretion of GAPDH in E. tarda EIB202 and mutants of several secretory pathways were analyzed by Western blot and ELISA. The levels of GAPDH in the supernatants of mutant library