CAJ | 학술논문

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2.9μg/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。
목적：연제사문씨균 invA 기인중조질립，위분자생물학방법쾌속검측사문씨균제공질립표준。방법통과PCR확증목적편단，련접지pMD 18-T재체，전화대장간균DH5α，측서방법증실목적편단이성공중조，형광정량 PCR방법정성검측분석，채용 PicoGreen DNA분자형광정량방법대표준질립분자진행정치。결과 invA기인목적편단성공중조지 pMD 18-T재체상，형광정량 PCR결과현시제비중조질립표준위사문씨균핵산표준，중조질립표준적농도위2.9μg/mL。결론성공구건사문씨균invA기인중조질립，위쾌속검측사문씨균전정료기출。
Objective To construct recombinant plasmid of invA gene as standard for the detection of Sal-monella 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 re-sults of the real-time quantitative PCR showed that the recombinant plasmid for the positive detection in Salmo-nella was validated. The concentration was 2.9μg/mL. Conclusion The recombinant plasmid of invA gene has been successfully constructed, which has established the foundation for the rapid detection of Salmonella.