摘要
参考GenBank上发表的H5亚型禽流感病毒(AIV)神经氨酸酶(NA)基因序列设计引物,PCR扩增NA基因,再将其克隆到原核表达载体pET-32a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61 kDa,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阳性血清发生特异性反应,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神经氨酸酶抗体具有良好的灵敏性。
The neuraminidase（ NA ）gene of Avian intluenza virus（AIV）subtype H5 was amplified by PCR according to the sequence published on GenBank. It was cloned into the expression vector PET-32a. The protein encoded by this gene was expressed in Escherichia coli strain BL21 （DE3）and was characterized by SDS-PAGE and western blotting analysis as a 61 kDa protein with immunogenicity. The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renenaturation. The results indicated that this protein could be specifically recognized by H5 subtype AIV antiserum. The assay has good specificity to H5 AIV. The ELISA result demonstrated that the purified protein could be used to detect the antibody against NA of AIV subtype H5N1 and had good sensitivity.
出处
《华北农学报》
CSCD
北大核心
2008年第5期107-110,共4页
Acta Agriculturae Boreali-Sinica
基金
国家“973”计划项目(2005CB523200)
