Physical and functional mapping of the transcriptional start sites of Plasmodium falciparum proliferating cell nuclear antigen
Physical and functional mapping of the transcriptional start sites of Plasmodium falciparum proliferating cell nuclear antigen
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DOI：
10.1016/0166-6851(96)02737-5
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发表时间：
1996-11-25
影响因子：
1.5
通讯作者：
Kilbey, BJ
中科院分区：
文献类型：
--
作者：
Horrocks, P;Kilbey, BJ
核糖核酸酶保护试验和引物延伸分析已经被用来定位PfPCNA编码序列翻译起始上游的960bp的主要转录起始点。第二个较小的地点位于上游40个BP处。在PfPCNAL5‘侧翼序列的不同修饰元件的转录控制下，将含有萤火虫荧光素酶报告基因的构建体瞬时导入红细胞内寄生虫阶段。这些实验确定了一个对启动子活性至关重要的470bp区域，该区域包含物理定位的转录起始点。此外，转录起始点上游的290至620个碱基之间的区域是有效启动子活性所必需的。
RNase protection assays and primer extension analysis have been used to locate a major transcription start site 960 bp upstream from the translational start of the PfPCNA coding sequence. A second, minor, site is situated a further 40 bp upstream. Intraerythrocytic parasite stages were transiently transfected with constructs containing a firefly luciferase reporter gene under the transcriptional control of variously modified elements of the PfPCNA 5' flanking sequence. These experiments identified a 470 bp region essential for promoter activity, which contains the physically mapped transcriptional start sites. In addition, a region between 290 and 620 bp upstream of the transcriptional start sites is required for efficient promoter activity.