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作 者:谢昆[1,2] 陶宏征[1] 王传铭[1] 李河[1] 林丽飞[1] 王栋[1]
机构地区:[1]红河学院生命科学与技术学院,蒙自661100 [2]云南省天然药物与化学生物学重点实验室,蒙自661100
出 处:《安徽农业大学学报》2009年第2期273-277,共5页Journal of Anhui Agricultural University
基 金:红河学院生物化学与分子生物学重点学科建设项目(071010);红河学院博硕科研启动项目(XS205026)共同资助
摘 要: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.
关 键 词:柔嫩艾美耳球虫 DNA疫苗 pEtK2-IFN-γ 构建
分 类 号:S852.723[农业科学—基础兽医学]
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