沙门氏菌invA基因重组质粒标准的构建

Construction of recombinant plasmid of invA gene of Salmonella

中文关键词:  沙门氏菌  重组质粒  invA基因

英文关键词:Salmonella  recombinant plasmid  invA gene

中文摘要:

      目的 研制沙门氏菌invA基因重组质粒, 为分子生物学方法快速检测沙门氏菌提供质粒标准。 方法 通过PCR扩增目的片段, 连接至pMD 18-T载体, 转化大肠杆菌DH5α, 测序方法证实目的片段已成功重组, 荧光定量PCR方法定性检测分析, 采用PicoGreen DNA分子荧光定量方法对标准质粒分子进行定值。结果 invA基因目的片段成功重组至pMD 18-T载体上, 荧光定量PCR结果显示制备重组质粒标准为沙门氏菌核酸标准, 重组质粒标准的浓度为2.9 μg/mL。结论 成功构建沙门氏菌invA基因重组质粒, 为快速检测沙门氏菌奠定了基础。

英文摘要:

      Objective To construct recombinant plasmid of invA gene as standard for the detection of Salmonella by molecular biology methods. Methods The target segment was amplified by PCR and cloned into pMD 18-T vector, then transformed into Escherichia coli DH5α. The recombinant plasmid was identified by se-quencing. The value of standard plasmid was defined by PicoGreen DNA fluorescence quantitative method. Results The target segment was successfully recombined into pMD 18-T vector with correct sequences. The results of the real-time quantitative PCR showed that the recombinant plasmid for the positive detection in Salmonella was validated. The concentration was 2.9 μg/mL. Conclusion The recombinant plasmid of invA gene has been successfully constructed, which has established the foundation for the rapid detection of Salmonella.