• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Aug 1983; 155(2): 847–853.
PMCID: PMC217758

Organization and subcloning of the dacA-rodA-pbpA cluster of cell shape genes in Escherichia coli.


The transducing bacteriophage lambda pBS10 carries a small cluster of Escherichia coli penicillin-binding protein/cell shape genes, including pbpA, rodA, and dacA. Deletion mapping and subcloning showed that these genes, and the gene for a cytoplasmic membrane protein of molecular weight 54,000, are located within a 5.6-kilobase region and are probably contiguous. The dacA gene, which codes for penicillin-binding protein 5, was cloned on a 1.5-kilobase fragment into a low-copy-number plasmid vector, but insertion into high-copy-number plasmids produced deleterious effects on bacterial growth, and the plasmids could not be stably maintained. The direction of transcription of dacA was determined. The rodA gene was cloned on a 1.6-kilobase fragment into both low- and high-copy-number plasmids, and the identification of its gene product is described in the accompanying paper (Stoker et al., J. Bacteriol. 155:854-859). The pbpA gene, which codes for penicillin-binding protein 2, was cloned on a 3.7-kilobase fragment in low-copy-number plasmids, but insertion of the fragment into high-copy-number plasmids resulted in deleterious effects on bacterial growth, and the plasmids could not be stably maintained.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bachmann BJ, Low KB. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. [PMC free article] [PubMed]
  • Broome-Smith JK, Spratt BG. Deletion of the penicillin-binding protein 6 gene of Escherichia coli. J Bacteriol. 1982 Nov;152(2):904–906. [PMC free article] [PubMed]
  • Cohen SN, Chang AC, Boyer HW, Helling RB. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240–3244. [PMC free article] [PubMed]
  • Daniels DL, de Wet JR, Blattner FR. New map of bacteriophage lambda DNA. J Virol. 1980 Jan;33(1):390–400. [PMC free article] [PubMed]
  • Herrero E, Jackson M, Bassford PJ, Sinden D, Holland IB. Insertion of a MalE beta-galactosidase fusion protein into the envelope of Escherichia coli disrupts biogenesis of outer membrane proteins and processing of inner membrane proteins. J Bacteriol. 1982 Oct;152(1):133–139. [PMC free article] [PubMed]
  • Levine A, Bailone A, Devoret R. Cellular levels of the prophage lambda and 434 repressors. J Mol Biol. 1979 Jul 5;131(3):655–661. [PubMed]
  • Markiewicz Z, Broome-Smith JK, Schwarz U, Spratt BG. Spherical E. coli due to elevated levels of D-alanine carboxypeptidase. Nature. 1982 Jun 24;297(5868):702–704. [PubMed]
  • Matsuhashi M, Tamaki S, Curtis SJ, Strominger JL. Mutational evidence for identity of penicillin-binding protein 5 in Escherichia coli with the major D-alanine carboxypeptidase IA activity. J Bacteriol. 1979 Jan;137(1):644–647. [PMC free article] [PubMed]
  • Nishimura Y, Suzuki H, Hirota Y, Park JT. A mutant of Escherichia coli defective in penicillin-binding protein 5 and lacking D-alanine carboxypeptidase IA. J Bacteriol. 1980 Jul;143(1):531–534. [PMC free article] [PubMed]
  • Parkinson JS, Huskey RJ. Deletion mutants of bacteriophage lambda. I. Isolation and initial characterization. J Mol Biol. 1971 Mar 14;56(2):369–384. [PubMed]
  • Pratt JM, Holland IB, Spratt BG. Precursor forms of penicillin-binding proteins 5 and 6 of E. coli cytoplasmic membrane. Nature. 1981 Sep 24;293(5830):307–309. [PubMed]
  • Reeve J. Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol. 1979;68:493–503. [PubMed]
  • Rogers HJ. Biogenesis of the wall in bacterial morphogenesis. Adv Microb Physiol. 1979;19:1–62. [PubMed]
  • Spratt BG. Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2999–3003. [PMC free article] [PubMed]
  • Spratt BG. Properties of the penicillin-binding proteins of Escherichia coli K12,. Eur J Biochem. 1977 Jan;72(2):341–352. [PubMed]
  • Spratt BG. Deletion of the penicillin-binding protein 5 gene of Escherichia coli. J Bacteriol. 1980 Dec;144(3):1190–1192. [PMC free article] [PubMed]
  • Spratt BG, Boyd A, Stoker N. Defective and plaque-forming lambda transducing bacteriophage carrying penicillin-binding protein-cell shape genes: genetic and physical mapping and identification of gene products from the lip-dacA-rodA-pbpA-leuS region of the Escherichia coli chromosome. J Bacteriol. 1980 Aug;143(2):569–581. [PMC free article] [PubMed]
  • Stoker NG, Fairweather NF, Spratt BG. Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene. 1982 Jun;18(3):335–341. [PubMed]
  • Stoker NG, Pratt JM, Spratt BG. Identification of the rodA gene product of Escherichia coli. J Bacteriol. 1983 Aug;155(2):854–859. [PMC free article] [PubMed]
  • Tamaki S, Matsuzawa H, Matsuhashi M. Cluster of mrdA and mrdB genes responsible for the rod shape and mecillinam sensitivity of Escherichia coli. J Bacteriol. 1980 Jan;141(1):52–57. [PMC free article] [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)


Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...