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鳗源嗜水气单胞菌β—溶血素基因的克隆及表达
引用本文:龚晖.鳗源嗜水气单胞菌β—溶血素基因的克隆及表达[J].水产学报,2003,27(2):124-130.
作者姓名:龚晖
作者单位:福建省农业科学院畜牧兽医研究所,福建,福州,350003
基金项目:国家863资助(2001AA622050),福建省科技项目资助(2000Z077)
摘    要:应用PCR技术,从1株鳗源嗜水气单胞菌ML316中扩增得到β 溶血素基因AHL316HEM,将AHL316HEM克隆到pGEM TEasyVector,经分析验证后重组到pcDNA3.0中,构建了重组质粒PDLH。转化重组质粒PDLH的大肠杆菌(Escherichiacoli)DH5α能在血琼脂培养皿中形成明显的β 溶血斑,重组菌纯化的胞外产物溶血价为1.28×104HU·mg-1,同时重组菌的胞外产物能被原始菌株的高免血清特异性地识别。结果证实,克隆到鳗源嗜水气单胞菌β 溶血素基因,重组质粒PDLH能够表达具有天然生物学活性的β 溶血素,为构建嗜水气单胞菌核酸疫苗奠定了基础。

关 键 词:鳗源嗜水气单胞菌  β-溶血素  基因克隆  基因表达  疫苗  鱼类  暴发性疾病  致病菌
文章编号:1000-0615(2003)02-0124-07
收稿时间:2014/3/18 0:00:00
修稿时间:2002年5月27日

Cloning and expressing of the β-hemolysin gene of Aeromonas hydrophila strain isolated from Anguilla anguilla
GONG Hui.Cloning and expressing of the β-hemolysin gene of Aeromonas hydrophila strain isolated from Anguilla anguilla[J].Journal of Fisheries of China,2003,27(2):124-130.
Authors:GONG Hui
Institution:Animal Husbandry and Veterinary Medicine Institute, Fujian Academy of Agricultural Sciences, Fuzhou350003, China
Abstract:With the technology of PCR, the completed ..-hemolysin gene AHL316HEM was obtained from the amplification of an Aeromonas hydrophila strain ML316 ( isolated from diseased Anguilla anguilla ) . The gene was cloned into pGEM-T Easy Vector. After analysis, the gene was inserted into pcDNA3. 0 vector to construct the recombinant plasmid PDLH. The E . coli DH5..w ith PDLH could lead to obvious ..-hemolysis plaque in blood-Agar. The hemolysis activity of crude purif ied transformant.. s extracellular products( ECP) was 1. 28 .. 104HU..mg- 1. Furthermore, the transformant.. s ECP could be recognized by ant-i serum which was raised against original bacterial strain. The above results demonstrated that the ..-hemolysin gene of ML316 had been cloned and the recombinant plasmid PDLH could express the product with native bio logical function. The success of cloning and expressing the ..-hemolysin gene of A . hydrophila will speed up the development of DNA vaccine against A . hydrophila.
Keywords:Aeromonas hydrophila      -hemolysin gene  cloning  expression
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