| 摘要: |
| 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。 |
| 关键词: 鼠疫耶尔森氏菌 YscF抗原 克隆表达 |
| DOI： |
| 分类号: |
| 基金项目: |
|
| Expression and identification of YscF antigen gene of Yersinia pestis in E.coli |
|
|
| Abstract: |
| 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. |
| Key words: Yersinia pestis,YscF antigen,cloning expression |
