鸡柔嫩艾美耳球虫免疫调节型DNA疫苗pEtK2-IFN-γ的构建

Construction of immunoregulating DNA vaccine against Eimeria tenella

利用DNA重组技术,分别构建pVAX1-pEtK2、pcDNA4.0(c)-pEtK2、pVAX1-pEtK2-IFN-γ、pcDNA4.0(c)-pEtK2-IFN-γDNA疫苗载体。重组质粒肌肉注射14日龄鸡,1周后肌肉注射部位组织切除和裂解,通过RT-PCR,Western-blotting检测保护性抗原在肌肉中表达情况。结果表明,鸡柔嫩艾美耳球虫pEtK2表面抗原基因和鸡IFN-γ基因均能在注射部位肌肉成功表达,这为鸡球虫DNA疫苗的研制奠定了基础。

pVAXI-pEtK2, pcDNA4.0 （ c ）-pEtK2, pVAXI-pEtK2-IFN-γ and pcDNA4. 0 （ c ）-pEtK2-IFN-γ DNA vectors were constructed by the DNA recombinant technique. After identificatied by restriction enzyme digestion,the recombinant DNA plasmids were injected into chest muscle of 14-day-old chicken respectively, then the muscle tissue samples were treated, and expression of these genes and their products were detected by RT-PCR and western blotting respectively. The result indicated that both pErK2 gene and IFN-γ gene were successfully expressed in chicken chest muscle tissue, and it laid foundation for further study on DNA vaccine of coecidian.