【摘要】 采用PCR技术从重组质粒pMel-NP扩增得到了禽流感病毒(AIV)A/Goose/Guang-dong/1/96(H5N1)株的核蛋白(NP)基因,将其亚克隆至真核表达载体pVAX1上。将提取的pVAX-NP阳性克隆转染Hela细胞,48 h后经间接免疫荧光试验和Western-blotting检测,NP基因在Hela细胞中已成功瞬时表达。表达产物的分子质量约为57 ku,它与H5N1亚型AIV多克隆抗血清有较好的免疫反应。更多还原

【Abstract】 The NP gene of the AIV A/Goose/Guangdong/1/96(H5N1) strain was amplified from a recombinant plasmid pMel-NP by PCR and subcloned into an eukaryotic expression vector pVAX1 and the recombinant vector was designated pVAX-NP.The high quality pVAX-NP was prepared and the Hela cells were transfected.The recombinant vector pVAX-NP was successfully expressed in the cells.Indirect immunofluorescence assay and Western-blotting analysis showed that the nucleoprotein was expressed transiently in the Hela cells,which was 57ku in molecular weight,and had good immunologic reactivity with the polyclonal antiserum of AIV H5N1 subtype.更多还原