Protein expression in Escherichia coli minicells containing recombinant plasmids specifying trimethoprim-resistant dihydrofolate reductases.

Journal of bacteriology ·Vol. 141 ·No. 2 ·1980-02-00 ·Pages 779-85

Fling ME, Elwell LP

Abstract

Deoxyribonucleic acid fragments containing the structural genes for several trimethoprim-resistant dihydrofolate reductases from naturally occurring plasmids were inserted into small cloning vehicles. The genetic expression of these hybrid plasmids was studied in purified Escherichia coli minicells. The type I dihydrofolate reductase, encoded by plasmid R483 and residing within transposon 7 (Tn7), had a subunit molecular weight of 18,000. The type II dihydrofolate reductase, specified by plasmid R67, had a subunit molecular weight of 9,000. These two enzymes were antigenically distinct in that anti-type II dihydrofolate reductase (R67) antibody did not cross-react with the type I (R483) protein. The trimethoprim-resistant reductase specified by plasmid R388 had a subunit molecular weight of about 10,500 and was immunologically related to the type II (R67) enzyme. A 9,000 subunit of the dihydrofolate encoded by the transposition element Tn402 was also antigenically related to the R67 reductase.

MeSH Terms

Bacterial Proteins/biosynthesis Escherichia coli/drug effects,genetics,metabolism Molecular Weight Peptide Biosynthesis Plasmids R Factors Recombination, Genetic Tetrahydrofolate Dehydrogenase/genetics,metabolism Trimethoprim/pharmacology

Chemicals

Bacterial Proteins Trimethoprim Tetrahydrofolate Dehydrogenase

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Fling M E

Elwell L P

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25 references, click to expand

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Article Info

Journal

Journal of bacteriology

Abbr.

J Bacteriol

ISSN

0021-9193

DOI

10.1128/jb.141.2.779-785.1980

Published

1980-02-00

Pages

779-85

Language

English

Region

United States

NLM ID

2985120R

PMCID

PMC293688

Subset

External Links

PubMed source DOI: 10.1128/jb.141.2.779-785.1980 PMC Full Text View journal details

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Protein expression in Escherichia coli minicells containing recombinant plasmids specifying trimethoprim-resistant dihydrofolate reductases.

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