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.
出处
《中国兽医学报》
CAS
CSCD
北大核心
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)
