作　　者： ; ;

机构地区： 暨南大学生命科学技术学院

出　　处： 《生物技术通报》 2011年第10期126-129,共4页

摘　　要： 构建人β-酪蛋白GAS基序荧光素酶报告基因表达载体。合成3个连续重复的人β-酪蛋白启动子中干扰素γ活化序列(gamma-activated sequence,GAS)并将其克隆至pGL3-Promoter荧光素酶报告基因载体。测序鉴定构建的载体,然后将其转染入人胚肺成纤维细胞(human embryonic lung fibroblasts,HELF)中检测荧光素酶的表达活性。测序结果表明,3×GAS序列正确;双报告基因系统检测荧光素酶活性表明,构建的报告基因具有启动子活性。成功构建的人β-酪蛋白GAS基序荧光素酶报告基因表达载体,为进一步研究干扰素γ诱导的JAK-STAT信号通路提供了必要的试验材料。 It was to construct the luciferase reporter gene vector of human β-casein GAS motif.Three duplicates of gamma-activated sequence（GAS） of human β-casein promoter was made,which was cloned into pGL3-Promoter luciferase reporter gene vector.The constructed vector was confirmed by sequencing and then transfected into human embryonic lung fibroblasts（HELF） to detect luciferase activity.The sequencing result indicated that the 3×GAS sequence was correct,and the luciferase activity by Dual-Luciferase Reporter Assay System demonstrated that the constructed vector displayed the promoter activity.The human β-casein GAS motif luciferase reporter gene vector was successfully constructed,which would play an important role of the IFN-γ induced JAK-STAT signaling pathway in further study.

关 键 词： 酪蛋白 基序 荧光素酶 报告基因

领　　域： [生物学]