鳗源嗜水气单胞菌β—溶血素基因的克隆及表达
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福建省农业科学院畜牧兽医研究所

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国家高技术研究发展计划(863计划),福建省科技攻关项目


Cloning and expressing of the β-hemolysin gene of Aeromonas hydrophila strain isolated from Anguilla anguilla
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Animal Husbandry and Veterinary Medicine Institute, Fujian Academy of Agricultural Sciences, Fuzhou350003, China

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    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-T Easy Vector,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(Escherichia coli)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.

    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.

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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):

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  • 收稿日期:2014-03-18
  • 最后修改日期:2014-03-18
  • 录用日期:2014-03-18
  • 在线发布日期: 2014-03-18
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