【摘要】 为构建表达鸡传染性支气管炎病毒(IBV)主要免疫原S1蛋白的重组腺病毒,本研究以复制缺陷型人5型腺病毒为载体,以IBVCK/CH/LHLJ/04V株S1基因为外源插入靶基因,通过细菌内同源重组法构建了一株稳定表达IBVS1蛋白(90ku)的重组腺病毒,命名为rAdV-S1。通过PCR鉴定、间接免疫荧光及western blot检测证实S1蛋白在重组腺病毒中获得表达。对构建的重组腺病毒和亲本腺病毒的生长动力学分析表明,该重组病毒的毒价为108.25TCID50/mL,并且两者在生长动力学方面无显著差异。本研究为动物试验和该重组腺病毒免疫特性的研究奠定基础。更多还原

【Abstract】 The S1 subunit of spike protein(90 ku) of infectious bronchitis virus(IBV) is the main immunodominant protein which can induce virus neutralization and haemagglutination-inhibition antibodies.In this study,a recombinant replication-defective human 5 adenovirus(rAdV-S1) that expresses S1 gene from IBV strain CK/CH/LHLJ/04V was constructed with viral titer of 108.25 TCID50/mL in HEK293.Western blot and indirect immunofluorescence assay indicated that the S1 gene was expressed in HEK293 cells infected with rAdV-S1.The growth dynamics indicated that there were no statistically differences between the rAdV-S1 and parental adenovirus.The rAdV-S1 could be sued to further study for the immunogenicity in chickens.更多还原