PMID: 6405380Feb 1, 1983Paper

Chromogenic identification of genetic regulatory signals in Bacillus subtilis based on expression of a cloned Pseudomonas gene

Proceedings of the National Academy of Sciences of the United States of America
M M ZukowskiJ P Lecocq

Abstract

A method to isolate fragments of DNA that promote gene expression in Bacillus subtilis is described. The system is based on production of catechol 2,3-dioxygenase [CatO2ase; catechol:oxygen 2,3-oxidoreductase (decyclizing), EC 1.13.11.2] encoded by the Pseudomonas putida TOL plasmid gene xylE. The gene was transferred to aB. subtilis/Escherichia coli plasmid vector to construct pTG402. Although xylE is functionally expressed in E. coli, CatO2ase is not detected in B. subtilis unless a fragment of DNA capable of promoting gene expression is ligated into a cleavage site on pTG402 upstream from xylE. Fragments of chromosomal DNA from B. subtilis, Bacillus licheniformis, Bacillus pumilus, and E. coli are shown to promote xylE gene expression in B. subtilis. The special feature of the system is the method of detection: colonies of cells that express xylE become yellow within seconds after selection plates are sprayed with catechol, a colorless substrate that is converted by CatO2ase to the yellow product, 2-hydroxymuconic semialdehyde. The complete nucleotide sequence of xylE is presented. Strong complementarity between the ribosome binding site and 16S rRNA suggests that xylE mRNA translation in B. subtilis may commence at the same...Continue Reading

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Citations

Sep 1, 1990·Applied Microbiology and Biotechnology·C M HooijmansK C Luyben
Sep 30, 2008·Applied Microbiology and Biotechnology·Elena G AbalakinaYurgis A V Yomantas
Feb 15, 2011·Applied Microbiology and Biotechnology·Joanne HothersallChristopher M Thomas
Aug 29, 2012·Antonie van Leeuwenhoek·Theresa Siegl, Andriy Luzhetskyy
Jun 1, 1986·Analytical Biochemistry·J DomenJ G Schoenmakers
Dec 30, 1994·Gene·E Clark, G Cirvilleri
Nov 1, 1995·Research in Microbiology·J C SirardA Fouet
Nov 8, 2011·Journal of Microbiological Methods·Aneta A BartosikGrażyna Jagura-Burdzy
Mar 27, 2001·FEMS Microbiology Letters·G González-CerónL Servín-González

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