Weir J P, Moss B
Virology. 1987 May;158(1):206-10. doi: 10.1016/0042-6822(87)90254-6.
Nine recombinant vaccinia viruses that contain overlapping segments of the putative promoter region of the vaccinia virus thymidine kinase (TK) gene linked to DNA coding for the prokaryotic enzyme chloramphenicol acetyltransferase (CAT) were constructed. In each case, the RNA start site and 5 bp of DNA downstream were retained. No significant difference in CAT expression occurred as the deletion was extended from 352 to 32 bp before the RNA start site. Deletion of a further 10 bp, however, led to complete cessation of early promoter activity. Primer extension analysis of the 5' ends of the transcripts verified that the natural TK RNA start site was still used when only 32 bp of upstream DNA remained. Loss of early promoter activity was previously found when deletions were extended from 31 to 24 bp before the RNA start site of another vaccinia gene that is expressed constitutively throughout infection (M.A. Cochran, C. Puckett, and B. Moss, 1985, Proc. Natl. Acad. Sci. USA 82, 19-23). Sequence similarities in the promoter regions of these two genes were noted.
构建了9种重组痘苗病毒，这些病毒含有痘苗病毒胸苷激酶（TK）基因推定启动子区域的重叠片段，这些片段与编码原核酶氯霉素乙酰转移酶（CAT）的DNA相连。在每种情况下，都保留了RNA起始位点和下游5个碱基对的DNA。当缺失从RNA起始位点前352个碱基对扩展到32个碱基对时，CAT表达没有显著差异。然而，再缺失10个碱基对会导致早期启动子活性完全停止。对转录本5'端的引物延伸分析证实，当仅保留上游DNA的32个碱基对时，仍然使用天然的TK RNA起始位点。之前发现，当缺失从另一个在整个感染过程中持续表达的痘苗基因的RNA起始位点前31个碱基对扩展到24个碱基对时，早期启动子活性丧失（M.A. Cochr an、C. Puckett和B. Moss，1985年，《美国国家科学院院刊》82，19 - 23）。注意到这两个基因启动子区域的序列相似性。
