鸡痘病毒表达载体的构建Ⅰ、鸡痘病毒282E_4株基因组PstⅠ、HindⅢ酶切片段的克隆及酶切分析

Construction of Recombiant Fowlpor As Expressing Vectors Ⅰ .Cloning and Mapping of fowlo virus Genomic DNA Pst Ⅰ. Hind Ⅲ Restricted Fragment

本文介绍了鸡痘病毒２８２Ｅ４株基团组ＰｓｔⅠ片段、ＨｉｎｄⅢ片段的克隆和鉴定，以及部分克隆的限制性内切酶分析。鸡痘病毒基因组经酶切、克隆分别得到３４９个、２７７个白色菌落，经质粒电泳、菌落杂交筛选分别得到２８６、２３４个重组菌落，再经酶切、斑卢、杂交鉴定，证明了所克隆的确为鸡痘病毒的ＰｓｔⅠ、ＨｉｎｄⅢ片段。最后经单、双酶切分析，得到了５个ｐＦＰ、５个ｐＦＨ克隆的物理图谱。

This paper describes the identification and cloning of Pat Ⅰ .Hind Ⅲ fragment of the genome of FPV 282E4 strair, and the RE analysis of some clores. FPV genome were digested and clored in result of 349 and 277 of white colones obtained respectively. According to the resultS of Agarose gel electrophoresis mobility and colonies hybridization. We screened out 286 of pFP. 234 of pFH recombinants. The recombinants were further identifed by dot blot and digestion of pFP, pFH with Pst Ⅰ.Hind Ⅲ. Analysed by RE, a total ten, 5 of pFP and 5 of pFH. virus specific recombinants plasmids were mapped by double-digestion method.