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PMID: 2160940 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Identification of phosphate starvation-inducible genes in Escherichia coli K-12 by DNA sequence analysis of psi::lacZ(Mu d1) transcriptional fusions.

Journal of bacteriology ·Vol. 172 ·No. 6 ·1990-06-00 ·Pages 3191-200

Metcalf WW, Steed PM, Wanner BL

Abstract

Twenty-four independent phosphate starvation-inducible (psi) transcriptional fusions made with Mu d1(lacZbla) were analyzed by sequencing the psi::lacZ(Mu d1) chromosomal junctions by using DNAs amplified with the polymerase chain reaction or mini-Mu cloning. Our DNA sequence analysis showed that the MuR DNA in Mu d1 has an unexpected structure that is comprised of 104 bases of MuR DNA in the form of a large inverted repeat, which we denoted Mu d1-R. Also, Mu d1s in the phoA and phn (psiD) loci of the phosphate regulon showed regional specificities for the insertion sites despite the randomness of Mu d1 insertions into the genome as a whole. Gene products or open reading frames were identified for seven unknown psi::lacZ(Mu d1) transcriptional fusions by searching DNA data bases with the sequences adjacent and upstream of the Mu d1s. One psiC::lacZ(Mu d1) lies in the ugpB gene of the ugpBAEC operon, which encodes a periplasmic sn-glycerol-3-phosphate-binding protein; two psiQ::lacZ(Mu d1)s lie in the gltB gene, and one psiQ::lacZ(Mu d1) lies in the gltD gene of the gltBDF operon, encoding the large and small subunits of glutamate synthase, respectively; and the psi-51::lacZ(Mu d1) lies in the glpB gene of the glpABC operon, which codes for the anaerobically regulated glycerol-3-phosphate dehydrogenase. psiE and psiF::lacZ(Mu d1)s lie in uncharacterized open reading frames near the xylE and phoA genes, respectively. Six other psi::lacZ(Mu d1)s lie in yet unreported Escherichia coli sequences.

MeSH Terms
Bacteriophage mu/genetics Base Sequence Cloning, Molecular DNA Transposable Elements DNA, Bacterial/analysis Escherichia coli/genetics Genes, Bacterial Lac Operon Molecular Sequence Data Operon Phosphates/metabolism Polymerase Chain Reaction Promoter Regions, Genetic Transcription, Genetic
Chemicals
DNA Transposable Elements DNA, Bacterial Phosphates
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Metcalf W W
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
Steed P M
Wanner B L
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.6.3191-3200.1990
Published
1990-06-00
Pages
3191-200
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC209124
Subset
IM
Grants
NIAID NIH HHS · AI27713 · United States
NIGMS NIH HHS · GM35392 · United States
Databases
GENBANK
M3328, M33723, M33724, M33725, M33726, M33727, M33728, M33729, M33730, M33731, M33732, M33733, M33734, M33735, M33736, M33737, M33738, M33739, M33740, M33741, M33742, M33743, M33744, M33745
External Links
PubMed source DOI: 10.1128/jb.172.6.3191-3200.1990 PMC Full Text View journal details
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