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。
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.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2007年第11期19-23,共5页
Journal of Northwest A&F University(Natural Science Edition)
