Eaton Heather E, Metcalf Julie, Brunetti Craig R
Trent University, Department of Biology, Peterborough, ON, Canada.
Can J Microbiol. 2008 Jun;54(6):483-8. doi: 10.1139/w08-033.
The conserved sequence element (CSE) is a highly conserved 42-bp poxvirus sequence that can function as a poxvirus promoter element. The CSE is composed of 2 repeats, each containing the highly conserved late poxvirus promoter sequence TAAAT. To define the location of the nucleotides critical for promoter function, polymerase chain reaction was carried out using primers that inserted modified versions of the CSE upstream of the green fluorescent protein (GFP), and the constructs were transiently transfected into cells by using GFP levels as a measure of promoter function. The results of this analysis revealed that the second TAAAT sequence, but not the first TAAAT sequence, is critical for promoter function of the CSE. Furthermore, deletion of half of the intervening sequence, i.e., from 10 to 5 nt, increases the promoter strength of the CSE as compared with the wild-type CSE. These results indicate the potential of this novel poxvirus promoter for driving high levels of gene expression.
保守序列元件（CSE）是一段高度保守的42个碱基对的痘病毒序列，可作为痘病毒启动子元件发挥作用。CSE由2个重复序列组成，每个重复序列都包含高度保守的痘病毒晚期启动子序列TAAAT。为了确定对启动子功能至关重要的核苷酸位置，使用引物进行聚合酶链反应，这些引物将CSE的修饰版本插入绿色荧光蛋白（GFP）上游，并且通过使用GFP水平作为启动子功能的衡量指标，将构建体瞬时转染到细胞中。该分析结果表明，第二个TAAAT序列而非第一个TAAAT序列对CSE的启动子功能至关重要。此外，与野生型CSE相比，删除一半的间隔序列（即从10到5个核苷酸）可增强CSE的启动子强度。这些结果表明这种新型痘病毒启动子具有驱动高水平基因表达的潜力。
