Wilkens K, Rüger W
Arbeitsgruppe Molekulare Genetik, Ruhr-Universität Bochum, Germany.
Plasmid. 1996 Mar;35(2):108-20. doi: 10.1006/plas.1996.0013.
We report on the construction of promoter probe vector pKWIII, useful in cloning and analyzing strong promoters for Escherichia coli RNA polymerase. Also T4 early promoters that proved to be difficult to clone with other vectors could be tested. The promoter activities obtained with this convenient and nonradioactive system largely correspond to those determined by pulse-labeling of transcripts in the same system. Results with well-characterized control promoters are in good agreement with values given by other authors. We present relative activities of several early promoters of phage T4 and compare these to promoter activities of other phages. Sorting the T4 promoters according to strength suggests the importance of distinct sequence elements to promoter functioning. They are centered around positions -52, -42, and -15.
我们报道了启动子探针载体pKWIII的构建，该载体可用于克隆和分析大肠杆菌RNA聚合酶的强启动子。此外，那些被证明难以用其他载体克隆的T4早期启动子也能够得到检测。通过这个便捷且非放射性的系统所获得的启动子活性，在很大程度上与在同一系统中通过转录本脉冲标记所确定的活性相符。使用特征明确的对照启动子所得到的结果，与其他作者给出的值高度一致。我们展示了噬菌体T4几个早期启动子的相对活性，并将其与其他噬菌体的启动子活性进行比较。根据强度对T4启动子进行排序表明，不同的序列元件对启动子功能具有重要性。它们集中在-52、-42和-15位附近。
