RT－cDNA克隆－PCR法获取犬瘟热病毒融合蛋白基因（F）

Cloning of the Gene Encoding Fusion Protein of Canine Distemper Virus Using RT-cDNA cloning──PCR Technique

纯化鸡胚成纤维细胞培养的犬瘟热病毒（ＣａｎｉｎｅＤｉｓｔｅｍｐｅｒＶｉｒｕｓ，ＣＤＶ），获得病毒基因组ＲＮＡ后，反转录合成双链病毒Ｆ基因ｃＤＮＡ。将此双链ｃＤＮＡ平端插入ＰＵＣ１９质粒ＳａｍⅠ位点构建重组质粒，进行ｃＤＮＡ克隆。以重组克隆质粒为模板ＰＣＲ扩增，获得ＣＤＶ全长Ｆ基因。将此Ｆ基因插入表达载体ＰＢＶ２２０，在大肠杆菌中表达，通过对表达产物的最终鉴定，可确认所获片段为ＣＤＶ全长Ｆ基因．

The gene encoding fusion protein of Canine Distemper Virus (CDV) was prepared from cDNA derived from virus genome RNA extracted from purified virus using RT-cDNA cloning -PCR and identified by restriction enzyme digestion analysis, PCR, Dot hybridization, In situ hybridization and the detection of induced protein of the cloned gene in E. colt. DH5a strain.