传染性支气管炎病毒(IBV)S1基因的真核表达及免疫原性检测

Eukaryotic Expression and Immunogenicity Detection of Avian Infectious Bronchitis Virus(IBV) S1 Gene

为了探讨S1基因在DNA免疫防制传染性支气管炎(IB)中的免疫原性,将IBVZ株S1基因插入到真核表达载体pcDNA3.1/V5-His-TOPO中,构建了真核表达质粒pcDNAIBVZS。在脂质体作用下将pcDNAIBVZS转染CEF细胞,用间接免疫荧光试验检测,在CEF细胞中可见有大量S1蛋白表达。将重组质粒以100μg/只的剂量肌注免疫SPF雏鸡,经间接ELISA试验检测二免前后的血清,结果显示,pcDNAIBVZS基因可在SPF鸡体内诱导相应抗体的产生,具有特定的免疫原性。

To assess the probability of S1 gene DNA vaccine in controlling IB.The IBV S1 gene was inserted into eukaryotic expression vector pcDNA3.1/V5-His-TOPO.A eukaryotic expression recombinant plasmid pcDNAIBVZS,containing start codon and stop codon,was constructed.The recombinant plasmids were transfected into CEF cells with the help of liposome and detected by indirect immuno-fluorescence experiment.Results demonstrated that S1 protein could be expressed in a large amout in CEF cells.The pcDNAIBVZS were injected into muscles of one-day old SPF chickens at a dose of 100 μg.Two weeks post injection,the serums of immuned chickens were collected to indirect ELISA,the test demonstrated that the IBVS1 gene DNA vaccine plasmids could induce the specific antibodies in chickens and had the good immunogenicity.