[Identification of transcriptional regulatory sequences of yigP gene in Escherichia coli].
OBJECTIVE
We investigated the promoter activity of yigP gene and analyzed its transcriptional regulatory sequence.


METHODS
We cloned promoter fragment into promoter probe plasmid which has a reporter gene lacZ, thus the promoter activity could be measured by detecting beta-galactosidase activity. Then, the promoter region was minimized by cloning different truncated promoter fragments into promoter probe plasmid. Using site-directed mutagenesis technology, we introduced site mutations in some key sequences and investigated their effects on promoter activity.


RESULTS
The promoter region of yigP gene was determined, and also the -10 region and -35 region were identified. Meanwhile, a negative regulatory sequence just upstream of yigP promoter was discovered, and our results show that these sequences have important influence on the transcriptional regulation.


CONCLUSIONS
We identified the transcriptional regulatory sequences of yigP gene, and this facilitated our understanding of gene transcription.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. wangyi495@126.com
Yi
Y
Wang
Jiang
J
Ye
Huizhan
H
Zhang
D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics*; D005798:Genes, Bacterial; D017930:Genes, Reporter; D026901:Membrane Transport Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods; D011401:Promoter Regions, Genetic; D050436:Regulatory Elements, Transcriptional
D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins