重组鼠疫菌F1抗原在大肠杆菌中的表达及免疫原性分析

Expression and immunogenicity study on the recombinant F1antigen of Yersinia pestis in E.coli

利用基因重组技术,用pET42(b+)质粒在大肠杆菌DE3中表达鼠疫菌F1抗原。经分析rF1抗原基因序列与天然F1抗原结构基因序列完全一致,电泳扫描测其表达量为25%:W estern B lot结果表明,rF1抗原可与F1特异性抗体相互作用,具有天然F1抗原的活性。用镍离子亲和层析纯化rF1抗原免疫BALB/c小鼠,在其血清中可检测到高滴度的抗F1抗体。

F1 gene fragment of Yersinia pestis was amplified by PCR,then the F1 antigen was expressed in E. coli DE3. Analysis of DNA sequence was demonstrated that rF1 was sequence was completely identical with the expected. Expression was obtained in E. coli DE3 and the expression level of F1 Ag reached to 25% and Western blot showed as a single band with a molecular weight of 17.6 Kd. The target protein was purified by Ni^＋ affinity chromatography. The high anti-F1 response were detected in the serum of rF1 immunized BALB/e mice.