Expression and identification of YscF antigen gene of Yersinia pestis in E.coli
2007
Cui Ping | Yu Sanke | Luo Deyan
Chinese. 为了克隆表达鼠疫耶尔森氏菌的YscF抗原基因，并对其免疫原性进行初步研究，将PCR扩增的YscF基因连接到pMD-18T载体，测序正确后再将YscF基因连接到表达载体PET32a，构建PET32a-YscF重组质粒，并在大肠杆菌BL21(DE3)中诱导表达；诱导表达蛋白采用Ni2+亲和层析法纯化，Western blot检测其免疫原性。结果显示，28 ku的PET32a-YscF融合蛋白能被兔抗鼠的多抗识别，说明表达产物具有良好的抗鼠疫抗原特异性。
Show more [+] Less [-]English. YscF gene of Yersinia pestis was cloned and expressed, and preliminary study on immunogenicity of recombinant protein YscF was carried out. YscF gene was inserted into pMD-18T vector after sequence correction, and recombinant expression plasmid PET32a-YscF was constructed by inserting YscF into PET32a and induced with IPTG; the target protein was purified by Ni2+ affinity chromatography; immunogenicity was detected by Western blot. The results indicated that the 28 ku expressed protein could be recognized by Yersinia pestis positive serum as expected. It’s proved that recombinant YscF is a possible alternative or adjunct protective vaccine antigen against plague. So YscF is an important protective antigen of Y. pestis.
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