RRC ID 30052
Author Asoh S, Matsuzawa H, Matsuhashi M, Ohta T.
Title Molecular cloning and characterization of the genes (pbpA and rodA) responsible for the rod shape of Escherichia coli K-12: analysis of gene expression with transposon Tn5 mutagenesis and protein synthesis directed by constructed plasmids.
Journal J. Bacteriol.
Abstract Two cell shape-determining genes of Escherichia coli K-12, pbpA, the structural gene for penicillin-binding protein 2, and rodA, whose protein is unknown, were subcloned into plasmid vectors from the transducing phage lambda MAd lip24, which carries the lip-leuS region of the E. coli chromosome. Plasmids with restriction enzyme-created deletions or transposon Tn5 insertions were isolated, and studies of genetic complementation of these plasmids with chromosomal mutations were carried out. Thus, a physical and genetic map of the rodA-pbpA region was established. The genes rodA and pbpA lie side by side within a 4.4-kilobase-pair region. The size of the rodA gene has been shown to be between 0.86 and 1.6 kilobase pairs; such DNA would encode a protein with a molecular weight between 32,000 and 59,000. Since Tn5 mutagenesis of the rodA gene did not affect the expression of the pbpA gene and vice versa, the genes rodA and pbpA seem to have independent promoters. Analysis of the proteins synthesized from the constructed plasmids in maxicells revealed that the plasmid carrying the pbpA gene encoded penicillin-binding protein 2 and amplification of the protein occurred. The product of the rodA gene was not identified.
Volume 154(1)
Pages 10-6
Published 1983-4
PMID 6300030
PMC PMC217424
MeSH Bacterial Proteins / genetics Carrier Proteins / genetics Chromosome Deletion Chromosomes, Bacterial / analysis Cloning, Molecular* DNA Restriction Enzymes DNA Transposable Elements DNA, Bacterial / analysis Escherichia coli / genetics* Gene Expression Regulation Genes Genes, Bacterial* Hexosyltransferases* Muramoylpentapeptide Carboxypeptidase* Mutation Penicillin-Binding Proteins Peptidyl Transferases* Plasmids
IF 3.219
Times Cited 15
Prokaryotes E. coli ME8275