王瑞旋, 耿玉静, 冯娟, 王江勇. 杂色鲍哈维弧菌耐药质粒的鉴定和分析[J]. 南方水产科学, 2012, 8(2): 1-6. DOI: 10.3969/j.issn.2095-0780.2012.02.001
引用本文: 王瑞旋, 耿玉静, 冯娟, 王江勇. 杂色鲍哈维弧菌耐药质粒的鉴定和分析[J]. 南方水产科学, 2012, 8(2): 1-6. DOI: 10.3969/j.issn.2095-0780.2012.02.001
WANG Ruixuan, GENG Yujing, FENG Juan, WANG Jiangyong. Identification and analysis of resistant plasmid ofpathogenic bacteria Vibrio harveyi isolated from Haliotis diversicolor[J]. South China Fisheries Science, 2012, 8(2): 1-6. DOI: 10.3969/j.issn.2095-0780.2012.02.001
Citation: WANG Ruixuan, GENG Yujing, FENG Juan, WANG Jiangyong. Identification and analysis of resistant plasmid ofpathogenic bacteria Vibrio harveyi isolated from Haliotis diversicolor[J]. South China Fisheries Science, 2012, 8(2): 1-6. DOI: 10.3969/j.issn.2095-0780.2012.02.001

杂色鲍哈维弧菌耐药质粒的鉴定和分析

Identification and analysis of resistant plasmid ofpathogenic bacteria Vibrio harveyi isolated from Haliotis diversicolor

  • 摘要: 文章对引起杂色鲍(Hallotis diversicolor)肌肉萎缩症的病原菌哈维弧菌(Vibrio harveyi)质粒上的磺胺耐药基因进行研究,结果显示,该菌株对复方新诺明完全耐药,其质粒上含有sulⅡ耐药基因。接合转化试验显示该质粒具可转移性,可使受体菌对磺胺制剂复方新诺明产生耐药性,鉴定为耐药质粒,并测定了该耐药质粒的全基因组序列,序列总长为10 940 bp,初步分析显示有7个具有一定功能的ORF框。进一步构建重组表达质粒pRSET-A-sulⅡ,表达了目的蛋白(31 kDa)。

     

    Abstract: The pathogenic bacteria Vibrio harveyi discovered in abalone Haliotis diversicolor with withering syndrome is completely resistant to trimethoprim-sulfamethoxazole. This study identifies the resistance genes and their location in this bacterias plasmid. sulⅡwas detected in the plasmid with polymerase chain reaction. The conjugative transformation test shows that the plasmid is transferable and leads to the resistance of bacteria to sulfonamide. The complete sequence of the R plasmid is 10 940 bp long including 7 ORF which display biological function. sulⅡwas cloned into the multiple cloning sites pRSET-A(+) to build a recombinant positive plasmid. The SDS-PAGE indicates that the recombinant sulⅡprotein, in the form of inclusion body, is 31 kDa.

     

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