Inouye S, Asai Y, Nakazawa A, Nakazawa T
J Bacteriol. 1986 Jun;166(3):739-45. doi: 10.1128/jb.166.3.739-745.1986.
A DNA segment that promotes gene expression in Pseudomonas putida was identified in pTN8, a mutant plasmid of an RP4-TOL recombinant. A promoter on the segment was cloned with a promoter-probe vector containing the xylE gene of the TOL plasmid. The xylE gene was expressed under the control of the promoter, and the gene product catechol 2,3-dioxygenase was constitutively synthesized. As analyzed by an S1 nuclease protection assay, the amount of mRNA produced in P. putida was more than that in Escherichia coli. Fine S1 nuclease mapping and reverse transcriptase mapping revealed three tandem transcription start sites in both P. putida and E. coli. The nucleotide sequence preceding the transcription start sites was determined; a part of this sequence contained a sequence homologous to E. coli promoter sequences. A tentative consensus sequence for P. putida constitutive promoters is proposed.
在RP4-TOL重组体的突变质粒pTN8中鉴定出一段能促进恶臭假单胞菌基因表达的DNA片段。用含有TOL质粒xylE基因的启动子探针载体克隆了该片段上的一个启动子。xylE基因在该启动子的控制下表达，其基因产物儿茶酚2,3-双加氧酶可组成型合成。通过S1核酸酶保护试验分析，恶臭假单胞菌中产生的mRNA量比大肠杆菌中的多。精细的S1核酸酶图谱分析和逆转录酶图谱分析揭示了恶臭假单胞菌和大肠杆菌中均有三个串联的转录起始位点。确定了转录起始位点之前的核苷酸序列；该序列的一部分包含与大肠杆菌启动子序列同源的序列。提出了恶臭假单胞菌组成型启动子的暂定共有序列。
