van Delft J H, Mariñon B, Schmidt D S, Bosch L
Department of Biochemistry, University of Leiden, The Netherlands.
Nucleic Acids Res. 1987 Nov 25;15(22):9515-30. doi: 10.1093/nar/15.22.9515.
Signals setting the level of transcription of the tRNA-tufB operon have been studied by deletion mapping. TufB transcription was measured in vivo with plasmid-borne tRNA-tufB:galk operon fusions. Removal of the sequences from -133 to -58 with respect to the transcription start point, results in a 90% decrease of tufB transcription. This demonstrates the presence of a region, upstream of the tRNA-tufB promoter, that enhances the expression of the operon. DNA fragments bearing this upstream activator region do not display an abnormal electrophoretic mobility, as has been observed for the rrnB P1 upstream activator. Deletions starting in the first tRNA gene and directing towards tufB reveal at least two sites that influence tufB transcription. One signals transcription termination in the intergenic region between thrT and tufB. The other may be involved in antitermination. Possible mechanisms underlying antitermination and termination are considered in the light of the nucleotide sequence.
已通过缺失作图法研究了设定tRNA - tufB操纵子转录水平的信号。利用质粒携带的tRNA - tufB:galk操纵子融合体在体内测量TufB转录。相对于转录起始点,去除从 - 133至 - 58的序列会导致tufB转录减少90%。这证明在tRNA - tufB启动子上游存在一个区域,该区域可增强操纵子的表达。携带此上游激活区的DNA片段并未显示出异常的电泳迁移率,这与rrnB P1上游激活剂的情况不同。从第一个tRNA基因开始并指向tufB的缺失揭示了至少两个影响tufB转录的位点。一个位点在thrT和tufB之间的基因间隔区发出转录终止信号。另一个位点可能参与抗终止作用。根据核苷酸序列考虑了抗终止和终止的潜在机制。