Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1990 Jul 25;18(14):4139–4142. doi: 10.1093/nar/18.14.4139

On the use of T7 RNA polymerase transcripts for physical investigation.

A A Szewczak 1, S A White 1, D T Gewirth 1, P B Moore 1
PMCID: PMC331170  PMID: 1696001

Abstract

A few years ago we made some observations which raised questions about the accuracy with which T7 RNA polymerase transcribes templates in vitro, and the suitability of its in vitro products for biophysical study (1). The experiments described below demonstrate that there is no reason for concern; the products of T7 RNA polymerase transcription in vitro are as suitable for biophysical characterization as RNAs synthesized in vivo. It is likely that aggregation involving the transcribed portions of the T7 RNA polymerase promoter caused our initial observations.

Full text

PDF
4139

Selected References

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

  1. Davanloo P., Rosenberg A. H., Dunn J. J., Studier F. W. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2035–2039. doi: 10.1073/pnas.81.7.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dunn J. J., Studier F. W. Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol. 1983 Jun 5;166(4):477–535. doi: 10.1016/s0022-2836(83)80282-4. [DOI] [PubMed] [Google Scholar]
  3. Gewirth D. T., Moore P. B. Effects of mutation on the downfield proton nuclear magnetic resonance spectrum of the 5S RNA of Escherichia coli. Biochemistry. 1987 Sep 8;26(18):5657–5665. doi: 10.1021/bi00392a012. [DOI] [PubMed] [Google Scholar]
  4. Gewirth D. T., Moore P. B. Exploration of the L18 binding site on 5S RNA by deletion mutagenesis. Nucleic Acids Res. 1988 Nov 25;16(22):10717–10732. doi: 10.1093/nar/16.22.10717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hall K. B., Sampson J. R., Uhlenbeck O. C., Redfield A. G. Structure of an unmodified tRNA molecule. Biochemistry. 1989 Jul 11;28(14):5794–5801. doi: 10.1021/bi00440a014. [DOI] [PubMed] [Google Scholar]
  6. Kime M. J., Moore P. B. Physical evidence for a domain structure in Escherichia coli 5 S RNA. FEBS Lett. 1983 Mar 7;153(1):199–203. doi: 10.1016/0014-5793(83)80147-1. [DOI] [PubMed] [Google Scholar]
  7. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Milligan J. F., Groebe D. R., Witherell G. W., Uhlenbeck O. C. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. Nucleic Acids Res. 1987 Nov 11;15(21):8783–8798. doi: 10.1093/nar/15.21.8783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Puglisi J. D., Wyatt J. R., Tinoco I., Jr A pseudoknotted RNA oligonucleotide. Nature. 1988 Jan 21;331(6153):283–286. doi: 10.1038/331283a0. [DOI] [PubMed] [Google Scholar]
  10. Sampson J. R., Uhlenbeck O. C. Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1033–1037. doi: 10.1073/pnas.85.4.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sen D., Gilbert W. A sodium-potassium switch in the formation of four-stranded G4-DNA. Nature. 1990 Mar 29;344(6265):410–414. doi: 10.1038/344410a0. [DOI] [PubMed] [Google Scholar]
  12. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  13. Varani G., Wimberly B., Tinoco I., Jr Conformation and dynamics of an RNA internal loop. Biochemistry. 1989 Sep 19;28(19):7760–7772. doi: 10.1021/bi00445a036. [DOI] [PubMed] [Google Scholar]
  14. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 1983;100:468–500. doi: 10.1016/0076-6879(83)00074-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES