Mapping a eukaryotic promoter: a DNA sequence required for in vivo expression of adenovirus pre-early functions.
Mapping a eukaryotic promoter: a DNA sequence required for in vivo expression of adenovirus pre-early functions.
复制标题
绘制真核启动子图谱：体内表达腺病毒早期功能所需的 DNA 序列。
DOI：
10.1073/pnas.78.3.1381
复制
发表时间：
1981
影响因子：
11.1
通讯作者：
Berk,AJ
中科院分区：
文献类型：
--
作者：
Osborne,TF;Schell,RE;Burch-Jaffe,E;Berget,SJ;Berk,AJ
本研究定义了腺病毒2早期前区体内表达所需的mRNA帽位点上游的DNA序列。将腺病毒2的前早期区和侧翼序列克隆到大肠杆菌质粒pBR 322中。构建缺乏上游病毒DNA序列部分的质粒衍生物。设计了一种测定法来测试这些质粒DNA补充具有前早期区域缺失的腺病毒5突变体的能力。在mRNA帽位点上游保留至少38个碱基对的质粒具有与原始质粒相似的互补活性，原始质粒含有229个碱基对的上游病毒序列。然而，从帽位点上游保留23个或更少碱基对的病毒序列的质粒具有显著降低的互补活性。这些结果表明，在mRNA帽位点上游23和38个碱基对之间的腺病毒2序列的一部分是表达早期前区所必需的。该间隔包括Goldberg-Hogness盒状序列T-A-T-T-T-A-T-A。
This study defines a DNA sequence upstream from the mRNA cap site required for in vivo expression of the adenovirus 2 pre-early region. The adenovirus 2 pre-early region and flanking sequences were cloned in Escherichia coli plasmid pBR322. Derivatives of the plasmid lacking portions of the upstream viral DNA sequence were constructed. An assay was devised to test the ability of these plasmid DNAs to complement an adenovirus 5 mutant with a deletion in the pre-early region. Plasmids that retained at least 38 base pairs upstream from the mRNA cap site had complementing activity similar to that of the original plasmid, which contains 229 base pairs of upstream viral sequence. However, plasmids retaining 23 or fewer base pairs of viral sequence upstream from the cap site had significantly reduced complementing activity. These results indicate that a portion of the adenovirus 2 sequence between 23 and 38 base pairs upstream from the mRNA cap site is required for expression of the pre-early region. This interval includes the Goldberg-Hogness box-like sequence T-A-T-T-T-A-T-A.