Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1984 Apr 11;12(7):3219–3234. doi: 10.1093/nar/12.7.3219

Two improved promoter sequences for the beta-lactamase expression arising from a single base-pair substitution.

S T Chen, R C Clowes
PMCID: PMC318740  PMID: 6326054

Abstract

A mutation in the transposon Tn2660 derived from the plasmid R6K, and resulting in an approximate tenfold increase in penicillin resistance is shown to be a single GC to AT substitution 177 base pairs 'upstream' of the initiation codon of the structural beta-lactamase (bla) gene. This substitution leads to the transcription of two new mRNAs which can be ascribed to the creation of two new overlapping promoter sequences. All the sequences (450bp) examined in the wild-type Tn2660 are identical to those reported in Tn3.

Full text

PDF
3219

Images in this article

3226Image
on p.3226
3228Image
on p.3228

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ambler R. P., Scott G. K. Partial amino acid sequence of penicillinase coded by Escherichia coli plasmid R6K. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3732–3736. doi: 10.1073/pnas.75.8.3732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Brosius J., Cate R. L., Perlmutter A. P. Precise location of two promoters for the beta-lactamase gene of pBR322. S1 mapping of ribonucleic acid isolated from Escherichia coli or synthesized in vitro. J Biol Chem. 1982 Aug 10;257(15):9205–9210. [PubMed] [Google Scholar]
  5. Chaconas G., van de Sande J. H. 5'-32P labeling of RNA and DNA restriction fragments. Methods Enzymol. 1980;65(1):75–85. doi: 10.1016/s0076-6879(80)65012-5. [DOI] [PubMed] [Google Scholar]
  6. Chiang S. J., Clowes R. C. Intramolecular transposition and inversion in plasmid R6K. J Bacteriol. 1980 May;142(2):668–682. doi: 10.1128/jb.142.2.668-682.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chiang S. J., Clowes R. C. Recombination between two TnA transposon sequences oriented as inverse repeats is found less frequently than between direct repeats. Mol Gen Genet. 1982;185(1):169–175. doi: 10.1007/BF00333809. [DOI] [PubMed] [Google Scholar]
  8. Chiang S. J., Jordan E., Clowes R. C. Intermolecular and intramolecular transposition and transposition immunity in Tn3 and Tn2660. Mol Gen Genet. 1982;187(2):187–194. doi: 10.1007/BF00331116. [DOI] [PubMed] [Google Scholar]
  9. Clowes R. C., Holmans P. L., Chiang S. J. Intramolecular transposition of beta-lactamase sequence and related genetic rearrangements. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):167–171. doi: 10.1101/sqb.1981.045.01.026. [DOI] [PubMed] [Google Scholar]
  10. Datta N., Kontomichalou P. Penicillinase synthesis controlled by infectious R factors in Enterobacteriaceae. Nature. 1965 Oct 16;208(5007):239–241. doi: 10.1038/208239a0. [DOI] [PubMed] [Google Scholar]
  11. Dempsey W. B., Willetts N. S. Plasmid co-integrates of prophage lambda and R factor R100. J Bacteriol. 1976 Apr;126(1):166–176. doi: 10.1128/jb.126.1.166-176.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dennis P. P., Nomura M. Regulation of the expression of ribosomal protein genes in Escherichia coli. J Mol Biol. 1975 Sep 5;97(1):61–76. doi: 10.1016/s0022-2836(75)80022-2. [DOI] [PubMed] [Google Scholar]
  13. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heffron F., McCarthy B. J., Ohtsubo H., Ohtsubo E. DNA sequence analysis of the transposon Tn3: three genes and three sites involved in transposition of Tn3. Cell. 1979 Dec;18(4):1153–1163. doi: 10.1016/0092-8674(79)90228-9. [DOI] [PubMed] [Google Scholar]
  15. Heffron F., Sublett R., Hedges R. W., Jacob A., Falkow S. Origin of the TEM-beta-lactamase gene found on plasmids. J Bacteriol. 1975 Apr;122(1):250–256. doi: 10.1128/jb.122.1.250-256.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Holmans P. L., Clowes R. C. Transposition of a duplicate antibiotic resistance gene and generation of deletions in plasmid R6K. J Bacteriol. 1979 Feb;137(2):977–989. doi: 10.1128/jb.137.2.977-989.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kingsbury D. T., Helinski D. R. Temperature-sensitive mutants for the replication of plasmids in Escherichia coli: requirement for deoxyribonucleic acid polymerase I in the replication of the plasmid ColE 1 . J Bacteriol. 1973 Jun;114(3):1116–1124. doi: 10.1128/jb.114.3.1116-1124.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kolter R. Replication properties of plasmic R6K. Plasmid. 1981 Jan;5(1):2–9. doi: 10.1016/0147-619x(81)90073-1. [DOI] [PubMed] [Google Scholar]
  20. Kontomichalou P., Mitani M., Clowes R. C. Circular R-factor molecules controlling penicillinase synthesis, replicating in Escherichia coli under either relaxed or stringent control. J Bacteriol. 1970 Oct;104(1):34–44. doi: 10.1128/jb.104.1.34-44.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matthew M., Hedges R. W. Analytical isoelectric focusing of R factor-determined beta-lactamases: correlation with plasmid compatibility. J Bacteriol. 1976 Feb;125(2):713–718. doi: 10.1128/jb.125.2.713-718.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  23. Oka A., Nomura N., Morita M., Sugisaki H., Sugimoto K., Takanami M. Nucleotide sequence of small ColE1 derivatives: structure of the regions essential for autonomous replication and colicin E1 immunity. Mol Gen Genet. 1979 May 4;172(2):151–159. doi: 10.1007/BF00268276. [DOI] [PubMed] [Google Scholar]
  24. Pribnow D. Bacteriophage T7 early promoters: nucleotide sequences of two RNA polymerase binding sites. J Mol Biol. 1975 Dec 15;99(3):419–443. doi: 10.1016/s0022-2836(75)80136-7. [DOI] [PubMed] [Google Scholar]
  25. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  26. Russell D. R., Bennett G. N. Characterization of the beta-lactamase promoter of pBR322. Nucleic Acids Res. 1981 Jun 11;9(11):2517–2533. doi: 10.1093/nar/9.11.2517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Selzer G., Som T., Itoh T., Tomizawa J. The origin of replication of plasmid p15A and comparative studies on the nucleotide sequences around the origin of related plasmids. Cell. 1983 Jan;32(1):119–129. doi: 10.1016/0092-8674(83)90502-0. [DOI] [PubMed] [Google Scholar]
  28. Som T., Tomizawa J. Regulatory regions of ColE1 that are involved in determination of plasmid copy number. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3232–3236. doi: 10.1073/pnas.80.11.3232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  31. von Gabain A., Belasco J. G., Schottel J. L., Chang A. C., Cohen S. N. Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts. Proc Natl Acad Sci U S A. 1983 Feb;80(3):653–657. doi: 10.1073/pnas.80.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES