Abstract
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
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Broome-Smith J. K., Spratt B. G. Deletion of the penicillin-binding protein 6 gene of Escherichia coli. J Bacteriol. 1982 Nov;152(2):904–906. doi: 10.1128/jb.152.2.904-906.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen S. N., Chang A. C., Boyer H. W., Helling R. B. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240–3244. doi: 10.1073/pnas.70.11.3240. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Daniels D. L., de Wet J. R., Blattner F. R. New map of bacteriophage lambda DNA. J Virol. 1980 Jan;33(1):390–400. doi: 10.1128/jvi.33.1.390-400.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herrero E., Jackson M., Bassford P. J., Sinden D., Holland I. B. 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. doi: 10.1128/jb.152.1.133-139.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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. doi: 10.1016/0022-2836(79)90014-7. [DOI] [PubMed] [Google Scholar]
- Markiewicz Z., Broome-Smith J. K., Schwarz U., Spratt B. G. Spherical E. coli due to elevated levels of D-alanine carboxypeptidase. Nature. 1982 Jun 24;297(5868):702–704. doi: 10.1038/297702a0. [DOI] [PubMed] [Google Scholar]
- Matsuhashi M., Tamaki S., Curtis S. J., Strominger J. L. 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. doi: 10.1128/jb.137.1.644-647.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nishimura Y., Suzuki H., Hirota Y., Park J. T. 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. doi: 10.1128/jb.143.1.531-534.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parkinson J. S., Huskey R. J. Deletion mutants of bacteriophage lambda. I. Isolation and initial characterization. J Mol Biol. 1971 Mar 14;56(2):369–384. doi: 10.1016/0022-2836(71)90471-2. [DOI] [PubMed] [Google Scholar]
- Pratt J. M., Holland I. B., Spratt B. G. Precursor forms of penicillin-binding proteins 5 and 6 of E. coli cytoplasmic membrane. Nature. 1981 Sep 24;293(5830):307–309. doi: 10.1038/293307a0. [DOI] [PubMed] [Google Scholar]
- Reeve J. Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol. 1979;68:493–503. doi: 10.1016/0076-6879(79)68038-2. [DOI] [PubMed] [Google Scholar]
- Rogers H. J. Biogenesis of the wall in bacterial morphogenesis. Adv Microb Physiol. 1979;19:1–62. doi: 10.1016/s0065-2911(08)60197-6. [DOI] [PubMed] [Google Scholar]
- Spratt B. G., 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. doi: 10.1128/jb.143.2.569-581.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spratt B. G. Deletion of the penicillin-binding protein 5 gene of Escherichia coli. J Bacteriol. 1980 Dec;144(3):1190–1192. doi: 10.1128/jb.144.3.1190-1192.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spratt B. G. 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. doi: 10.1073/pnas.72.8.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spratt B. G. Properties of the penicillin-binding proteins of Escherichia coli K12,. Eur J Biochem. 1977 Jan;72(2):341–352. doi: 10.1111/j.1432-1033.1977.tb11258.x. [DOI] [PubMed] [Google Scholar]
- Stoker N. G., Fairweather N. F., Spratt B. G. Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene. 1982 Jun;18(3):335–341. doi: 10.1016/0378-1119(82)90172-x. [DOI] [PubMed] [Google Scholar]
- Stoker N. G., Pratt J. M., Spratt B. G. Identification of the rodA gene product of Escherichia coli. J Bacteriol. 1983 Aug;155(2):854–859. doi: 10.1128/jb.155.2.854-859.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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. doi: 10.1128/jb.141.1.52-57.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]