不同海洋弧菌中5类溶血素基因的分布及其与溶血活性和磷脂酶活性的相关性  

Distribution of five kinds of haemolysin genes in different vibrios and their correlation with activities of haemolytic and phospholipase

作　　者：王淑娴[1] 张晓华[1] 孙铂光[1] 陈吉祥[1] 

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《中国水产科学》2007年第4期570-578,共9页Journal of Fishery Sciences of China

基　　金：国家自然科学基金资助项目(30371119);教育部新世纪优秀人才支持计划(NCET-04-0645)资助项目

摘　　要：用57株海洋弧菌,包括26株弧菌标准菌株、20株哈维氏弧菌、11株副溶血弧菌(从不同宿主和不同地理环境中分离得到),用PCR法合成5类相应的地高辛标记的溶血素基因探针,利用其进行Southern Blot,检测这5类溶血素基因在57株弧菌中的分布。结果显示,在57海洋株弧菌中,含有TDH、HlyA、TLH、δ-VPH和HLX溶血素基因的菌株分别为2株、2株、49株、3株和30株。另外,1株霍氏格里蒙菌(Grimontia hollisae)和1株副溶血弧菌(V.parahaemolyticus)中分别含有2个TDH溶血素基因。用鱼血平板和卵磷脂平板检测57株弧菌的溶血活性和磷脂酶活性,结果表明,弧菌溶血活性和磷脂酶活性与TLH溶血素基因具有显著相关性,与另外4类溶血素基因的关系不明显。Vibrios are normal inhabitants in aquatic environments and very common in marine and estuarine habitats Many studies have reported that Vibrio species are pathogenic to marine shrimps and fish. Haemolysin, which is an exotoxin that attacks blood cell membranes and causes cell rupture, is arguably the most widely distributed toxin among pathogenic vibrios. Haemolysis may result from the enzymatic activities demonstrated by some species of bacteria, including phospholipase C of Pseudomonas aeruginosa and phos- pholipase D in Photobacterium damselae ssp. damselae. However, many other haemolysins act by formingpores in the cytoplasmic membrane of erythrocytes. In many cases, the pore-forming activity of haemolysin is not restricted to erythrocytes, but extends to a wide range of other cell types including mast ceils, neu- trophils, and polymorphonuclear cells, and enhances virulence by causing tissue damage. Currently, five representative haemolysin families were found in Vibrio sp., including the TDH （thermostable direct haemolysin） family of V. parahaemolyzicus, the HlyA （E1 Tor haemolysin） family of V. cholerae, the TLH （thermolabile haemolysin） family of V. parahaemolyticus, the δ-VPH （thermostable haemolysin） family of V. parahaemolyticus and the HLX family of V. cholerae O1. Each type of haemolysin gene was aligned using ClustalW （http：//www. ebi. ac. uk/clustalW/）,and five pairs of PCR primers were designed from the conserved region of the haemolysin genes and amplified the haemolysin gene fractions of TLH, TDH, HlyA, δ-VPH and HLX. Five DIG-labelled haemolysin gene probes were synthesized by PCR amplification from total DNA of different Vibrio strains, and were used to detect the distributions of the five haemolysin genes among 57 Vibrio strains by Southern Blot. The results showed that 2 Vibrio strains had TDH haemolysin gene;2 strains had HlyA haemolysin gene;49 strains had TLH haemolysin gene;3 strains had δ-VPH haemolysin gene and 30 strains had HLX haemolysin gene. From these res

关 键 词：弧菌 溶血素基因 溶血活性 磷脂酶活性 

分 类 号：S941[农业科学—水产养殖]