Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1981 Apr 24;9(8):1973–1989. doi: 10.1093/nar/9.8.1973

The construction of new vehicles for the cloning of transcription termination signals.

B E Enger-Valk, J van Rotterdam, P H Pouwels
PMCID: PMC326817  PMID: 6264402

Abstract

We have constructed new plasmids that can be used to clone transcription terminator containing DNA fragments between the first gene of the tryptophan (trp) operon and the tetracycline resistance (tet) gene. Both genes are under control of the trp promotor. Therefore the presence of a transcription termination signal on cloned fragments can be monitored by a decrease in expression of the tet gene. The plasmids contain cloning sites at different distances from the translation start signal. Consequently a cloned DNA fragment can be translated in the three possible reading frames, offering the opportunity to distinguish terminators from translation polarity (pseudo-terminators). The usefulness of the plasmids was shown by the cloning of the trp terminator and of a pseudo-terminator located in the trpB gene.

Full text

PDF
1973

Selected References

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

  1. Bolivar F., Rodriguez R. L., Betlach M. C., Boyer H. W. Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9. Gene. 1977;2(2):75–93. doi: 10.1016/0378-1119(77)90074-9. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  4. Crawford I. P., Nichols B. P., Yanofsky C. Nucleotide sequence of the trpB gene in Escherichia coli and Salmonella typhimurium. J Mol Biol. 1980 Oct 5;142(4):489–502. doi: 10.1016/0022-2836(80)90259-4. [DOI] [PubMed] [Google Scholar]
  5. Enger-Valk B. E., Heyneker H. L., Oosterbaan R. A., Pouwels P. H. Construction of new cloning vehicles with genes of the tryptophan operon of Escherichia coli as genetic markers. Gene. 1980 Apr;9(1-2):69–85. doi: 10.1016/0378-1119(80)90167-5. [DOI] [PubMed] [Google Scholar]
  6. Guarente L. P., Mitchell D. H., Beckwith J. Transcription termination at the end of the tryptophan operon of Escherichia coli. J Mol Biol. 1977 May 25;112(3):423–436. doi: 10.1016/s0022-2836(77)80190-3. [DOI] [PubMed] [Google Scholar]
  7. Hallewell R. A., Emtage S. Plasmid vectors containing the tryptophan operon promoter suitable for efficient regulated expression of foreign genes. Gene. 1980 Apr;9(1-2):27–47. doi: 10.1016/0378-1119(80)90165-1. [DOI] [PubMed] [Google Scholar]
  8. Mitchell D. H., Reznikoff W. S., Beckwith J. Genetic fusions that help define a transcription termination region in Escherichia coli. J Mol Biol. 1976 Mar 15;101(4):441–457. doi: 10.1016/0022-2836(76)90239-4. [DOI] [PubMed] [Google Scholar]
  9. Morse D. E., Yanofsky C. Polarity and the degradation of mRNA. Nature. 1969 Oct 25;224(5217):329–331. doi: 10.1038/224329a0. [DOI] [PubMed] [Google Scholar]
  10. Nichols B. P., Miozzari G. F., van Cleemput M., Bennett G. N., Yanofsky C. Nucleotide sequences of the trpG regions of Escherichia coli, Shigella dysenteriae, Salmonella typhimurium and Serratia marcescens. J Mol Biol. 1980 Oct 5;142(4):503–517. doi: 10.1016/0022-2836(80)90260-0. [DOI] [PubMed] [Google Scholar]
  11. Pannekoek H., Perbal B., Pouwels P. The specificity of transcription in vitro of the tryptophan operon of Escherichia coli. II. The effect of Rho factor. Mol Gen Genet. 1974;132(4):291–306. doi: 10.1007/BF00268570. [DOI] [PubMed] [Google Scholar]
  12. Roberts J. W. Termination factor for RNA synthesis. Nature. 1969 Dec 20;224(5225):1168–1174. doi: 10.1038/2241168a0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Sadler J. R., Betz J. L., Tecklenburg M., Goeddel D. V., Yansura D. G., Caruthers M. H. Cloning of chemically synthesized lactose operators. II. EcoRI-linkered operators. Gene. 1978 May;3(3):211–232. doi: 10.1016/0378-1119(78)90033-1. [DOI] [PubMed] [Google Scholar]
  15. Shimizu N., Hayashi M. In vitro transcription of the tryptophan operon integrated into a transducing phage genome. J Mol Biol. 1974 Apr 5;84(2):315–335. doi: 10.1016/0022-2836(74)90587-7. [DOI] [PubMed] [Google Scholar]
  16. Tait R. C., Rodriguez R. L., Boyer H. W. Altered tetracycline resistance in pSC101 recombinant plasmids. Mol Gen Genet. 1977 Mar 16;151(3):327–331. doi: 10.1007/BF00268797. [DOI] [PubMed] [Google Scholar]
  17. de Crombrugghe B., Mudryj M., DiLauro R., Gottesman M. Specificity of the bacteriophage lambda N gene product (pN): nut sequences are necessary and sufficient for antitermination by pN. Cell. 1979 Dec;18(4):1145–1151. doi: 10.1016/0092-8674(79)90227-7. [DOI] [PubMed] [Google Scholar]

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

RESOURCES