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目的在重组鸡痘病毒中表达传染性法氏囊病毒 VP2/VP243基因,并研究表达产物的保护性和免 疫原性。方法以 FPV 282E_4株 TK基因为侧翼,分别将鸡法氏囊病病毒(Infectious Bursal Disease Virus,IBDV) VP2 和VP243目的基因克隆到牛痘病毒A型包涵体(ATI)和痘苗病毒P7.5复合型启动子下游,构建成2个重组表达质 粒,命名为pUTALacVP和pUTA LacVPO。用这2个质粒转染CEF细胞,用X-gal染色法筛选出2株重组病毒vUTA lacVP2和 vUTALacVP0。用这 2株病毒免疫 1周龄 SPF鸡,以常规疫苗为对照。结果 ELISA、SDS-PAGE和 Western blot表明表达产物可与IBDV特异性多克隆抗体反应,并诱导鸡保护性抗体。在使用vvIBDV攻击前,对照组抗体水 平显著高于实验组。但攻击5 d后,实验组抗体水平明显升高。结论vUTALacVP2和 VPO在 CEF细胞中实现了高 效表达,但重组疫苗的保护率低于常规疫苗。
Objective To express the infectious bursal disease virus VP2 / VP243 gene in the recombinant fowlpox virus and study its protective and immunogenicity. Methods The TK gene of FPV 282E_4 was used as the flanks. The VP2 and VP243 genes of Infectious Bursal Disease Virus (IBDV) were cloned into vaccinia virus type A inclusion body (ATI) and vaccinia virus P7.5 complex Downstream of the promoter, two recombinant expression plasmids were constructed and named pUTALacVP and pUTA LacVPO. CEF cells were transfected with these two plasmids, and two recombinant viruses vUTA lacVP2 and vUTALacVP0 were screened by X-gal staining. One week-old SPF chickens were immunized with the two viruses, using the conventional vaccine as a control. Results ELISA, SDS-PAGE and Western blot showed that the expressed product reacted with IBDV-specific polyclonal antibody and induced protective antibody against chicken. Before using vvIBDV challenge, the control group antibody levels were significantly higher than the experimental group. However, after 5 days of challenge, the level of antibody in experimental group was significantly increased. Conclusions vUTALacVP2 and VPO are highly expressed in CEF cells, but the protective rate of recombinant vaccines is lower than that of conventional vaccines.