含有马立克氏病病毒糖蛋白B基因重组质粒的构建及其鉴定

CONSTRUCTION AND IDENTIFICATION OF RECOMBINANT PLASMID CONTAINING THE GLYCOPROTEIN B GENE OF MAREKS'DISEASE VIRUS

将马立克氏病病毒(MDV)GA 株基因文库中 BamH I I_3和 K_3克隆片段分别用 BamH I 加 Sal I 和 BamH I加 EcoR I 双酶切消化。含有编码 MDV 糖蛋白 B(gB)部分 DNA 的2.B kb、BamH I-Sal I 片段和1.1kb、BamH I-EcoR I 片段与 Sal I-EcoR I 消化的载体 pUR 222通过三聚体连接方式相连接。限制性核酸内切酶分析表明,重组质粒含有 MDV gB 基因,并且 MDV GA 株 gB 基因克隆在 EcoR I,Hind,Ⅲ,Xba I 和 Sal I 切点与 MDVRBIB 株相同。

The cloned BamH II_3 andK_3 fragments were excised from MDV Strain GA genomic library by doubledigestion with BamH I plus Sal I and with BamH I plus EcoR I,respectively.The 2.8 kb BamH I-Sal I fragment of I_3and the 1.1 kb BamH I-EcoR I fragment of K_3,both of which contain a part of DNA encoding the gB of MDV,wereligated with Sal I-EcoR I digested vector pUR222 by trimmer ligation.Endonuclease analysis showed that therecombinant plasmid contained the gB gene of MDV,and the gB gene clone of MDV strain GA was same as that of MDVstrain RB1B in EcoRⅠ,HindⅢ,Xba I and Sal I cleavage sites.