Inouye S, Nakazawa A, Nakazawa T
Gene. 1984 Sep;29(3):323-30. doi: 10.1016/0378-1119(84)90061-1.
The transcription initiation site of the xylDEGF operon on the TOL plasmid of Pseudomonas putida mt-2 was determined in P. putida and in Escherichia coli by S1 nuclease and reverse transcriptase mapping. The induced synthesis of mRNA started at the same start point in both P. putida and E. coli, although the amount of mRNA in E. coli cells was less than that in P. putida. The nucleotide sequence of the region surrounding the start point was also determined. The ribosome-binding site (RBS) complementary to the 3' end of the 16S rRNA of Pseudomonas aeruginosa and E. coli preceded the predicted start codon for the xylD gene. The consensus nucleotide sequence for E. coli promoters was not found in the region preceding the transcription start point. On the other hand, the sequences of the "-10" and the "-35" regions of the xylDEGF operon revealed some homology with the respective, previously determined sequences of the xylABC operon of the TOL plasmid.
通过S1核酸酶和逆转录酶图谱分析,在恶臭假单胞菌和大肠杆菌中确定了恶臭假单胞菌mt-2的TOL质粒上木糖DEGF操纵子的转录起始位点。尽管大肠杆菌细胞中的mRNA量少于恶臭假单胞菌中的mRNA量,但在恶臭假单胞菌和大肠杆菌中,mRNA的诱导合成均始于同一起始点。还确定了起始点周围区域的核苷酸序列。与铜绿假单胞菌和大肠杆菌16S rRNA 3'端互补的核糖体结合位点(RBS)位于木糖D基因预测的起始密码子之前。在转录起始点之前的区域未发现大肠杆菌启动子的共有核苷酸序列。另一方面,木糖DEGF操纵子的“-10”和“-35”区域的序列与TOL质粒木糖ABC操纵子各自先前确定的序列显示出一些同源性。