Ⅰ型马立克氏病病毒pp38和pp24基因的真核共表达

Co-expression and Construction of Eukaryote Plasmid of pp38 and pp24 of Marek′s Disease Virus

本研究将Ⅰ型超强毒MDVMd11株的pp38和pp24完整基因分别克隆到真核双表达载体pBudCE4.1中,在脂质体作用下将阳性克隆DNA转染CEF,通过间接免疫荧光试验(IFA)用单克隆抗体H19和鼠抗GST-pp24血清分别检测到了pp38和pp24基因的单独表达。然后将MDVMd11株的pp38和pp24完整基因同时克隆到载体pBudCE4.1中,在脂质体作用下将阳性克隆DNA转染CEF,通过IFA检测和用抗pp24多克隆血清进行West-ern-blotting试验检测到了PP38和PP24磷蛋白的共表达。

pp38 gene and pp24 gene of MDV Md11 strain were cloned into pBudCE 4.1 vector respectively. The pBudCE 4. 1-pp38 and pBudCE 4. 1-pp24 eukaryotic expressing plasmids were transfected into CEF respectively by lipofectamine reagent. 24, 48, 72 hours after transfection, the expression of pp38 was testified with Mab H19, and pp24 with the antiserum of pp24 expressed in E. coli. pp38 gene and pp24 gene were cloned into pBudCE 4. 1 vector which can express two different genes at the same time. After transfection,the co-expression of PP38 and PP24 was testified by IFA and by Western-blotting with Anti-GST-pp24 sera.