鸡贫血病毒全长基因串联体的构建

Construct of the full-length DNA concatemer of CAV

为了进一步研究鸡贫血病毒全长基因组感染性分子克隆和生物学特性,试验采用PCR方法,根据GenBank上已发表的鸡贫血病毒Cux-1株序列合成1对引物,利用高保真酶扩增鸡贫血病毒全长基因组DNA,并克隆到pB luescriptⅡSK(+)载体上,获得全长基因组DNA分子克隆。通过不完全酶切将2个全长基因连接,并进行酶切鉴定和测序分析。结果表明:2个全长基因为顺向连接,全长基因串联体构建成功。

To further study on the infectious DNA clone and biological characterization of CAV,a pair of primers was designed according to the full-length genomic sequence of Cux-1 strain of CAV based on GenBank data.The full-length genomic sequence of Cux-1 strain of CAV was amplified by PCR and cloned into pBluescript Ⅱ SK（＋） vector.The full-length DNA clone of CAV were obtained,two full-length DNA of CAV were linked by incomplete enzyme cutting.The results showed that the two full-length DNA of CAV were straight-forward linkage by restriction identification and sequencing analysis.It illustrated that the full-length DNA concatemer of CAV were successfully received.