犬2型腺病毒SY株E1区、E3区的克隆与序列分析

The cloning and sequence analysis of the E1 and E3 region of SY strain of CAV-2

根据犬2型腺病毒Toronto A26/61株的全基因序列物理图谱,确定E1、E3区位置,从CAV-2SY株的细胞培养物中提取了CAV-2全基因组DNA,经电泳检测其完整性并用多个限制性内切酶鉴定后,用EcoRⅠ和Kpn Ⅰ分别进行酶切,回收含E1区和E3区的相应片段,分别克隆到质粒pGEM-3zf和pBluescriptⅡKs(+)中,得到含E1区的重组质粒pGE1和含E3区的重组质粒pBE3-2。对获得的两个片段进行了序列测定和分析,结果表明,犬2型腺病毒SY株E1、E3基因的核苷酸序列与Toronto株的相应序列完全相同,其同源性均为100％。

The genome DNA of the SY strain of CAV-2 was extracted from the DK cells infected with CAV-2 and digested with EcoR I and Kpn I. The fragments containing the E1 and E3 regions were recovered and cloned into two plasmids and the two recombinant pasmids were named pG-E1 and pBE3-2 respectively. The E1 and E3 regions were sequenced and compared with the Toronto strain of CAV-2. The result showed that there were no differences between E1 and E3 regions sequence of the two strains of CAV-2.