tet启动子处的转录起始及突变的影响。

Harley C B, Lawrie J, Betlach M, Crea R, Boyer H W, Hedgpeth J

Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448.

Nucleic Acids Res. 1988 Aug 11;16(15):7269-85. doi: 10.1093/nar/16.15.7269.

We have identified the startpoint for transcription in vitro of the tetracycline resistance gene (tet) of pBR322 and several deletion and insertion mutations which alter tet promoter structure. Tetracycline resistance in host bacteria correlates qualitatively with the efficiency of DNA fragments from these plasmids to promote tet transcription in vitro. Only in active promoters could we find by computer analysis promoter structures in which the -10 and -35 sequences and the relative spacing of the two regions agree with consensus sequence determinants. These data support the current model of the E. coli promoter sequence. Two promoter mutants gave heterogeneous 5' termini with additional A residues not encoded by the DNA sequence.

我们已经确定了pBR322四环素抗性基因（tet）体外转录的起始点，以及几个改变tet启动子结构的缺失和插入突变。宿主细菌中的四环素抗性在质量上与来自这些质粒的DNA片段在体外促进tet转录的效率相关。只有在活性启动子中，我们通过计算机分析才能找到-10和-35序列以及这两个区域的相对间距与共有序列决定因素一致的启动子结构。这些数据支持了目前大肠杆菌启动子序列的模型。两个启动子突变体产生了异质性的5'末端，带有DNA序列未编码的额外A残基。