【摘要】 根据已发表的马立克氏病病毒(MDV)血清I型与鸡白介素2(IL2)DNA序列,设计两对引物,用PCR扩增出MDVgB基因和IL2基因,并克隆入pEGFPC1载体,构建了载体pC1gBIL。通过酶切分离外源基因表达盒,并将外源基因表达盒插入pTK2B质粒,成功构建了转移载体pTCgBIL。将转移载体pTCgBIL转染CEF细胞,培养36～48h后,经荧光显微镜观察,有荧光出现,证明该载体得到表达,并通过Westernblotting鉴定证明MDVgB和IL2的融合基因得到了有效的表达。更多还原

【Abstract】 Based on the published nucleotide sequence of MDV gB and interleukin-2(IL-2),the MDV gB and the interleukin-2 which designed and syntheisized by two pairs of primers respectively were cloned into the pEGFP-C1 vector and named pC1-gB-IL vector.Then gene experession cassette fragment was isolated from the pC1-gB-IL vector and was cloned into pTK2B vector.After the vector was transfected CEF cells and fostered 36 or 48 hours,the fused gene combined Marek′s disease virus gB and IL-2 was found to expressed in the transfected CEF cells by means of flourescent detection.Effective expression of the fused gene combined MDV gB and IL-2 was also testified by Western-blotting.更多还原