鸡痘病毒在哺乳细胞内复制的研究  

Recombinant fowl poxvirus vUTAL3CP1 multiplicationin in a mammalian cell line

作　　者：金明兰 金宁一[3] 鲁会军[3] 马鸣潇[3] 郑敏[3] 

机构地区：[1]吉林建筑工程学院松辽流域水环境教育部重点实验室,吉林长春130118 [2]国家兽医生物制品工程研究中心,江苏南京210014 [3]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2013年第3期336-340,共5页Chinese Journal of Veterinary Science

基　　金：江苏省博士后基金资助项目(080465)

摘　　要：将重组鸡痘病毒在1种禽类细胞和13种哺乳动物及人的细胞内培养。通过电镜观察可见重组鸡痘病毒在CEF、BHK、PK、BTC、Vero细胞内的复制,同时用PCR方法扩增出目的基因,Western-blotting检测到目的蛋白,验证是鸡痘病毒。试验表明鸡痘病毒在禽类、个别哺乳动物细胞中可以复制,但尚未发现在人的细胞内复制;在CEF中可以稳定传30代以上;在BHK内第1代可以观察到细胞病变(CPE);在PK、BTC、Vero内第2代可以观察到CPE,证明鸡痘病毒在个别哺乳动物细胞发生复制,但不能稳定遗传。Fowl poxviruses have many advantages and are being increasingly employed as recombi- nant vaccine vectors. One attractive feature is that while inserted transgenes are expressed in im- munologically favourable ways,fowl poxvirus infections of mammalian cells are believed to be a- bortive. The experimental evidence supporting this belief is, however, based on a limited number of mammalian cell-types and a few fowl poxvirus species. We evaluated two avian and 13 mamma- lian cell lines permissively to recombinant fowl poxvirus strains. Both avian cell lines were,as ex- pected,permissive for recombinant fowl poxvirus strains. However, by multiplication assays, we found to our surprise that syrian baby hamster kidney （BHK-21）,pig kidney （PK）,bovine testic- ular （BT）,bovine embryo kidney（MDBK）and bovine embryo trachea（EBTR）cells were equally permissive to recombinant strains. Results from electron microscopy of infected BHK-21, PK, BTC cells revealed viral morphogenesis proceeding to various forms of infectious viruses. These results were supported by the demonstration of fowl poxvirus specific late gene expression and fowl poxvirus specific DNA restriction pattern in BHK-21 infected cells. The results showed that the live attenuated virus adapted to the PK,BTC were safe as vaccine.

关 键 词：重组鸡痘病毒 BHK PK BT 

分 类 号：S852.65[农业科学—基础兽医学]