摘要
利用PCR方法从含禽流感病毒HA基因的质粒T-HA上扩增HA基因,再将其克隆到原核表达载体pET28a中,用E.coliBL21(DE3)原核表达,SDS-PAGE和Western-blot对表达蛋白进行鉴定。Western-blot结果表明,表达产物具有免疫学活性,蛋白的分子量约为60kD,位于包涵体中。包涵体经变性、复性处理,表达蛋白能与H5亚型AIV阳性血清特异性反应,具有良好的抗原性。ELISA检测结果表明,用此纯化蛋白作为包被抗原检测H5N1亚型AIV血凝素抗体具有良好的灵敏性。
The hemagglutinin （HA） gene of avian influenza virus （AIV） subtype H5N1 was amplified by PCR according to the sequence published on GenBank. Then, the PCR product was cloned into the expression vector pET28a and transformed into Escherichia coli strain BL21 （DEa） for the expression. The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renaturation. SDS--PAGE and western blotting analysis showed that the expressed protein is about 60 kD and could be specifically recognized by H5 subtype AIV antiserum. The ELISA results demonstrated that the purified protein could be used to detect the antibodies against HA protein of H5N1 subtype and represent high sensitivity.
出处
《河南农业科学》
CSCD
北大核心
2008年第8期127-130,共4页
Journal of Henan Agricultural Sciences
基金
国家"973"计划(2005CB523200)
