【摘要】 参照GenBank中登录的猪繁殖与呼吸综合征病毒(PRRSV)XH-GD株ORF6(M蛋白)基因序列,设计合成了一对引物,扩增PRRSV ORF6基因。将该基因克隆于转移质粒载体pFcDNA3.1(+)中,获得重组转移质粒pFcDNA3.1-ORF6并将其转化DH10Bac细胞,得到重组穿梭质粒Bacmid-ORF6,再将其转染昆虫细胞Sf9,获得重组杆状病毒rBacmid-ORF6,经PCR鉴定证实目的基因正确地插入到杆状病毒基因组的CMV启动子下游;经过间接免疫荧光试验(IFA)检测,M蛋白在Sf9细胞中得到了表达,而且表达的产物具有特异免疫学反应性。更多还原

【Abstract】 The ORF6(M protein) gene of PRRSV was amplified by PCR using a pair of primers based on the ORF6 sequences of a mutant PRRSV.The PCR product was cloned into the transfer vector pFcDNA3.1(+) and transformed into DH10Bac-host cells to produce recombinant shuttle plasmids Bacmid-ORF6,which was then used to transfect Sf9 insect cells.Indirect flurescent assay(IFA) confirmed that the ORF6 gene was expressed in Sf9 cells.The expression product could specially react to the sera of PRRSV.更多还原