Cordingley M G, Campbell M E, Preston C M
Nucleic Acids Res. 1983 Apr 25;11(8):2347-65. doi: 10.1093/nar/11.8.2347.
We have performed a functional analysis of DNA sequences upstream from the gene for IE mRNA3 of herpes simplex virus type 1. Nucleotide sequences involved in initiation and positive regulation of transcription have been defined by construction of specific deletions in vitro. Transcription was assayed in vivo by microinjection into Xenopus oocytes, or by introduction of plasmid DNA into tissue culture cells and measurement of transient expression. Three functional promoter elements have been defined: i) Sequences between -16 and -37 which are not essential for transcription but are required for accurate initiation. ii) Proximal promoter sequences which are sufficient for transcription initiation in the absence of upstream sequences. iii) Far-upstream promoter sequences (more than 108bp upstream) which increase transcription in oocytes, and contain positive regulatory sequences (-174 to -331) which respond strongly to a factor in the virus inoculum.
我们对1型单纯疱疹病毒IE mRNA3基因上游的DNA序列进行了功能分析。通过体外构建特定缺失片段，确定了参与转录起始和正调控的核苷酸序列。通过显微注射到非洲爪蟾卵母细胞中，或通过将质粒DNA导入组织培养细胞并测量瞬时表达，在体内检测转录情况。已确定了三个功能性启动子元件：i）-16至-37之间的序列，这些序列对转录不是必需的，但对准确起始是必需的。ii）近端启动子序列，在没有上游序列的情况下足以启动转录。iii）远上游启动子序列（上游超过108bp），可增加卵母细胞中的转录，并包含对病毒接种物中的一种因子有强烈反应的正调控序列（-174至-331）。
