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PMID: 2155195 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Mapping and molecular cloning of the phn (psiD) locus for phosphonate utilization in Escherichia coli.

Journal of bacteriology ·Vol. 172 ·No. 3 ·1990-03-00 ·Pages 1186-96

Wanner BL, Boline JA

Abstract

The Escherichia coli phn (psiD) locus encodes genes for phosphonate (Pn) utilization, for phn (psiD) mutations abolish the ability to use as a sole P source a Pn with a substituted C-2 or unsubstituted hydrocarbon group such as 2-aminoethylphosphonate (AEPn) or methylphosphonate (MPn), respectively. Even though the E. coli K-12 phosphate starvation-inducible (psi) phn (psiD) gene(s) shows normal phosphate (Pi) control, Pn utilization is cryptic in E. coli K-12, as well as in several members of the E. coli reference (ECOR) collection which are closely related to K-12. For these bacteria, an activating mutation near the phn (psiD) gene is necessary for growth on a Pn as the sole P source. Most E. coli strains, including E. coli B, are naturally Phn+; a few E. coli strains are Phn- and are deleted for phn DNA sequences. The Phn+ phn(EcoB) DNA was molecularly cloned by using the mini-Mu in vivo cloning procedure and complementation of an E. coli K-12 delta phn mutant. The phn(EcoB) DNA hybridized to overlapping lambda clones in the E. coli K-12 gene library (Y. Kohara, K. Akiyama, and K. Isono, Cell 50:495-508, 1987) which contain the 93-min region, thus showing that the phn (psiD) locus was itself cloned and verifying our genetic data on its map location. The cryptic phn(EcoK) DNA has an additional 100 base pairs that is absent in the naturally Phn+ phn(EcoB) sequence. However, no gross structural change was detected in independent Phn+ phn(EcoK) mutants that have activating mutations near the phn locus.

MeSH Terms
Cloning, Molecular DNA Transposable Elements Escherichia coli/genetics,metabolism Genes, Bacterial Genotype Gram-Negative Bacteria/genetics,metabolism Mutation Organophosphonates/metabolism Plasmids Restriction Mapping Transduction, Genetic
Chemicals
DNA Transposable Elements Organophosphonates
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wanner B L
Department of Biological Sciences, Purdue University, West Layfayette, Indiana 47907.
Boline J A
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.3.1186-1196.1990
Published
1990-03-00
Pages
1186-96
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC208583
Subset
IM
Grants
NIGMS NIH HHS · GM35392 · United States
External Links
PubMed source DOI: 10.1128/jb.172.3.1186-1196.1990 PMC Full Text View journal details
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