论文部分内容阅读
目的 研究MTS1基因 β启动子在急性T淋巴细胞白血病细胞株中的转录激活情况 ,鉴定 β启动子转录活性的功能片段区。方法 用DNA重组方法 ,构建MTS1基因 β启动子 3′端转录起始点相同 ,而 5′端序列不同的 7种pGL3重组质粒。用脂质体介导的基因瞬时转染法 ,将构建的重组质粒分别转染MTS1基因双等位缺失的Jurkat细胞株 ,检测pGL3重组质粒中荧光素酶报告基因的表达 ,观察 β启动子在Jurkat细胞中的激活情况及其基础转录活性片段区。 结果 成功构建了MTS1基因 β启动子 7种不同片段的重组质粒 ,它们在Jurkat细胞中均有转录活性 ,其中 0 .38kbSacⅡ SacⅠ酶切片段是MTS1基因 β启动子转录活性的基础片段。 结论 MTS1β启动子可在Jurkat细胞中被激活。
Objective To study the transcriptional activation of MTS1 β promoter in acute lymphoblastic leukemia cell line and to identify the functional fragment region of β promoter transcriptional activity. Methods Seven kinds of pGL3 recombinant plasmids with the same transcription start point at the 3 ’end of the β promoter of MTS1 gene and different sequences at the 5’ end were constructed by DNA recombination method. The recombinant plasmids were transfected into Jurkat cell line with double allele deletion of MTS1 gene by liposome-mediated gene transient transfection method to detect the expression of luciferase reporter gene in pGL3 recombinant plasmid. Activation in Jurkat cells and its basal transcriptional activity fragment. Results The recombinant plasmids of 7 different fragments of MTS1 gene promoter were successfully constructed and all of them were transcribed in Jurkat cells. The 0 .38kb Sac Ⅱ Sac Ⅰ fragment was the basic fragment of MTS1 gene β promoter transcriptional activity. Conclusion MTS1β promoter can be activated in Jurkat cells.