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目的验证嗜麦芽窄食单胞菌热休克蛋白HSP70(Dna K)的抗原性。方法以嗜麦芽窄食单胞菌K279a为模板,PCR扩增Dna K基因的全序列,将该序列连接到p ET-30a载体,转入大肠杆菌BL-21中,经1 mmol/L的IPTG 37℃诱导表达,目的蛋白纯化后进行Western印迹。结果与结论 Western印迹证实表达的重组蛋白能与鼠抗人HSP70单抗特异性结合,且与该菌的全菌免疫后血清有反应,而与免疫前血清无反应。嗜麦芽窄食单胞菌HSP70具有抗原性,可作为保护性抗原候选分子。
Objective To verify the antigenicity of heat shock protein HSP70 (Dna K) of Stenotrophomonas maltophilia. Methods The complete sequence of Dna K gene was amplified by PCR using the template of Stenotrophomonas maltophilia K279a as a template. The sequence was ligated into pET-30a vector and transformed into E. coli BL-21. After 1 mmol / L IPTG The expression was induced at 37 ° C, and the target protein was purified and subjected to Western blotting. RESULTS AND CONCLUSION Western blotting confirmed that the expressed recombinant protein could specifically bind to mouse anti-human HSP70 monoclonal antibody and reacted with whole-body immunized serum of this bacterium without reaction with pre-immune serum. Stenotrophomonas maltophilia HSP70 antigenicity, as a protective antigen candidate molecules.