摘要
将鸡痘病毒282E_4弱毒疫苗株的基因组分别进行Eco RI、Hind III和Bam HI酶切,以地高辛-dUTP标记澳大利亚鸡痘病毒疫苗株的TK基因作探针,进行Southern印迹杂交.结果发现,鸡瘟病毒282E_4弱毒株TK基因位于5.5kb Eco RI、3.7kb Hind III和4.3kb Bam HI酶切片段中.随后回收3.7kb Hind Ⅲ片段,克隆到pUC18质粒中.经Southern印迹杂交证实,已成功地克隆了我国鸡痘病毒282E_4弱毒株TK基因.
Genome of the 282E4 attenuated fowlpox virus was individually digested with restriction endonucleases Eco RI, Hind Ⅲ and Bam HI and separated by electrophoresis. The TK gene was located at 5. 5 kb Eco RI, 3. 7 kb Hind Ⅲ or 4. 3 kb Bam HI fragment respectively by Southern hybridization with the Digoxigenin labelled Australia TK gene, then the 3. 7 kb Hind Ⅲ fragment was recovered from agarose and cloned into pUC18 followed by Southern hybridization identification.
关键词
鸡痘病毒
TK基因
克隆
定位
fowlpox virus
thymidine kinase gene
clone
location
