鸡痘病毒282E4株Bam HI Fa,Fb,Fc片段非必需区筛选与序列分析

将亚克隆获得的分别含有鸡痘病毒（ Ｆ Ｐ Ｖ） Ｂａｍ Ｈ Ｉ相应片段的 ｐ Ｕ Ｆａ、ｐ Ｕ Ｆｃ 和 ｐ Ｕ Ｆｄ ３ 个质粒的单酶切位点，插入 Ｐ１１ Ｐ７．５ Ｌａｃ Ｚ报告基因盒，构建成 ｐ Ｕ Ｆａ１、ｐ Ｕ Ｆｃ１ 和 ｐ Ｕ Ｆｄ１ ３ 个重组载体质粒，在脂质体存在条件下，同 Ｆ Ｐ Ｖ 共同转染鸡胚成纤维细胞（ Ｃ Ｅ Ｆ），９６ ｈ 后在 Ｘｇａ１ 存在条件下挑选蓝色重组病毒空斑，经３ 次空斑纯化和传代，获得 １ 株可稳定表达 β半乳糖苷酶的重组病毒，并对该重组用质粒作了序列分析。

NER Selection and Nucleotide Sequence Analysis of Fa,Fb,Fc Bam HI Fragments in the FPV 282 E 4 Strain

The P11P7.5 LacZ reporter gene was inserted in the unique sites of Eco RV and Xho I in pUFa, pUFd and pUFc respectively. The resulting plasmids pUFa1, pUFc1 and pUFd1 were acquired, which were transfected with maternal FPV in CEF. After plaques selection,Passages and purifications under the condition of X gal staining, one recombinant FPV was found. And the nucleotide sequence analysis of pUFd was done so that it will be convinient for future application.