Brown J E, Klement J F, McAllister W T
Nucleic Acids Res. 1986 Apr 25;14(8):3521-6. doi: 10.1093/nar/14.8.3521.
Fragments of SP6 DNA generated by cleavage with Hpa II or Taq I were cloned into the Cla I site of pBR322 and the recombinant plasmids were screened for the presence of SP6 promoter activity by transcription in vitro with purified SP6 RNA polymerase. Three plasmids having promoter activity and small inserts of SP6 DNA were characterized. Hybridization studies showed that all three cloned promoters arose from different regions of the SP6 genome. Comparison of the consensus promoter sequence (5' ATTTAGGtgGACACTATAGAAGgaG 3') with the consensus sequences of promoters recognized by the T3 and T7 RNA polymerases reveals a common core sequence (5'-CACTA-3') extending from -7 to -3, as well as other features that may be important in selective promoter recognition by the phage RNA polymerases.
用Hpa II或Taq I切割产生的SP6 DNA片段被克隆到pBR322的Cla I位点，通过用纯化的SP6 RNA聚合酶进行体外转录来筛选重组质粒中SP6启动子活性的存在。对三个具有启动子活性且带有SP6 DNA小插入片段的质粒进行了表征。杂交研究表明，所有三个克隆的启动子都来自SP6基因组的不同区域。将共有启动子序列（5' ATTTAGGtgGACACTATAGAAGgaG 3'）与T3和T7 RNA聚合酶识别的启动子共有序列进行比较，发现了一个从-7到-3延伸的共同核心序列（5'-CACTA-3'），以及其他可能在噬菌体RNA聚合酶选择性识别启动子中起重要作用的特征。
