【摘要】 通过 PCR技术 ,以马立克氏病病毒 (MDV) RB1B株基因组 DNA为模板 ,扩增了包括 pp38基因及其启动子 -增强子和终止子在内的 2 2 0 0 bp的核酸片段。将该片段用 Sac 和 Sph 酶定向克隆到 p U C18质粒中 ,构建成重组质粒 ;将重组质粒转染鸡胚成纤维细胞 (CEF) ,用小鼠抗大肠杆菌表达的 pp38血清作间接免疫荧光试验 ,验证 pp38基因的表达。结果 ,在转染的细胞浆中看到了绿色的荧光 ,证实了该启动子的单向启动活性更多还原

【Abstract】 The (38 000) phosphoprotein gene of Marek′s disease virus RB1B strain with its promoter-enhancer and terminer were amplified through polymerase chain reaction(PCR).The (2 200) bp PCR product was cloned into pUC18 vector via restriction-enzyme SacⅠ and SphⅠ.The recombinant plasmid was transfected into chicken embryo fibroblast(CEF) by using liposome.Indirect fluorescent assay(IFA) with mouse-anti pp38 expressed in E.coli was used to identify the expression of pp38,and gray fluorescence on the transfected cells can be seen.It indicated that the promoter-enhancer could be used to construct a new eukaryotic expressing vector.更多还原