Improvement of an E. coli bioreporter for monitoring trace amounts of phenol by deletion of the inducible sigma54-dependent promoter.
An Escherichia coli bioreporter harboring the phenol-inducible mphK promoter (P(mphK)) from Acinetobacter calcoaceticus PHEA-2 fused to a beta-galactosidase gene (lacZ) and the regulator gene (mopR) of A. calcoaceticus NCIB8250 was constructed to detect phenol. P(mphK) containing three inverted repeats (IR1, IR2 and IR3) upstream of mphK was activated by the regulator MopR in the presence of phenol. Deletion analysis of P(mphK) revealed that IR2 and IR3 were essential for promoter activity, while IR1 was involved in transcriptional repression. The sensitivity of the bioreporter for the detection of phenol (0.1-5 microM) was improved by about 100% through deletion of IR1 in P(mphK).
Key Laboratory of Crop Biotechnology, Biotechnology Research Institute, Ministry of Agriculture, Chinese Academy of Agricultural Sciences, Beijing 100081, China. pengzixin1982@163.com
Zixin
Z
Peng
Yongliang
Y
Yan
Yuquan
Y
Xu
Masahiro
M
Takeo
Haiying
H
Yu
Zhonglin
Z
Zhao
Yuhua
Y
Zhan
Wei
W
Zhang
Min
M
Lin
Ming
M
Chen
D016954:Acinetobacter calcoaceticus / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins; D015374:Biosensing Techniques / Q000379:methods*; D004785:Environmental Pollutants / Q000032:analysis*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism; D017930:Genes, Reporter; D019800:Phenol / Q000032:analysis*; D011401:Promoter Regions, Genetic; D051877:RNA Polymerase Sigma 54 / Q000378:metabolism; D012680:Sensitivity and Specificity; D017384:Sequence Deletion; D015534:Trans-Activators; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D004785:Environmental Pollutants; D029968:Escherichia coli Proteins; C104241:MopR protein, Acinetobacter calcoaceticus; D015534:Trans-Activators; C497751:rpoN protein, E coli; D019800:Phenol; D051877:RNA Polymerase Sigma 54; D001616:beta-Galactosidase