Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1982 Aug 11;10(15):4609–4626. doi: 10.1093/nar/10.15.4609

Expression of a X. laevis tRNATyr gene in mammalian cells.

F A Laski, B Alzner-DeWeerd, U L RajBhandary, P A Sharp
PMCID: PMC321116  PMID: 6290987

Abstract

Expression of a X. laevis tRNATyr gene has been studied in mammalian cells. This tRNATyr gene has a 13 base intervening sequence adjacent to its anticodon. A fragment containing the tRNATyr gene was cloned into the late region of SV40. Cells infected with a recombinant virus stock vastly overproduce a tRNATyr that is properly spliced, processed and modified. It was also found that the X. laevis tRNATyr is identical or nearly identical to an endogenous tRNATyr of monkey kidney cells. The possibility of using the X. laevis tRNATyr gene to create an amber suppressor for mammalian cells is discussed.

Full text

PDF
4609

Images in this article

4615Image
on p.4615
4616Image
on p.4616
4618Image
on p.4618
4619Image
on p.4619

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., 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]
  2. Brambilla R., Rogg H., Staehelin M. Unexpected occurrence of an aminoacylated nucleoside in mammalian tRNATyr. Nature. 1976 Sep 9;263(5573):167–169. doi: 10.1038/263167a0. [DOI] [PubMed] [Google Scholar]
  3. Chu G., Sharp P. A. SV40 DNA transfection of cells in suspension: analysis of efficiency of transcription and translation of T-antigen. Gene. 1981 Mar;13(2):197–202. doi: 10.1016/0378-1119(81)90008-1. [DOI] [PubMed] [Google Scholar]
  4. Clarkson S. G., Kurer V., Smith H. O. Sequence organization of a cloned tDNA met fragment from Xenopus laevis. Cell. 1978 Jul;14(3):713–724. doi: 10.1016/0092-8674(78)90253-2. [DOI] [PubMed] [Google Scholar]
  5. Cremer K. J., Bodemer M., Summers W. P., Summers W. C., Gesteland R. F. In vitro suppression of UAG and UGA mutants in the thymidine kinase gene of herpes simplex virus. Proc Natl Acad Sci U S A. 1979 Jan;76(1):430–434. doi: 10.1073/pnas.76.1.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  7. Gilmore R. A. Super-suppressors in Saccharomyces cerevisiae. Genetics. 1967 Aug;56(4):641–658. doi: 10.1093/genetics/56.4.641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goff S. P., Berg P. Construction, propagation and expression of simian virus 40 recombinant genomes containing the Escherichia coli gene for thymidine kinase and a Saccharomyces cerevisae gene for tyrosine transfer RNA. J Mol Biol. 1979 Sep 25;133(3):359–383. doi: 10.1016/0022-2836(79)90398-x. [DOI] [PubMed] [Google Scholar]
  9. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  10. HAWTHORNE D. C., MORTIMER R. K. Super-suppressors in yeast. Genetics. 1963 Apr;48:617–620. doi: 10.1093/genetics/48.4.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  12. Hutchison C. A., 3rd, Phillips S., Edgell M. H., Gillam S., Jahnke P., Smith M. Mutagenesis at a specific position in a DNA sequence. J Biol Chem. 1978 Sep 25;253(18):6551–6560. [PubMed] [Google Scholar]
  13. Knapp G., Ogden R. C., Peebles C. L., Abelson J. Splicing of yeast tRNA precursors: structure of the reaction intermediates. Cell. 1979 Sep;18(1):37–45. doi: 10.1016/0092-8674(79)90351-9. [DOI] [PubMed] [Google Scholar]
  14. Kudo I., Leineweber M., RajBhandary U. L. Site-specific mutagenesis on cloned DNAs: generation of a mutant of Escherichia coli tyrosine suppressor tRNA in which the sequence G-T-T-C corresponding to the universal G-T-pseudouracil-C sequence of tRNAs is changed to G-A-T-C. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4753–4757. doi: 10.1073/pnas.78.8.4753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mulligan R. C., Howard B. H., Berg P. Synthesis of rabbit beta-globin in cultured monkey kidney cells following infection with a SV40 beta-globin recombinant genome. Nature. 1979 Jan 11;277(5692):108–114. doi: 10.1038/277108a0. [DOI] [PubMed] [Google Scholar]
  16. Müller F., Clarkson S. G. Nucleotide sequence of genes coding for tRNAPhe and tRNATyr from a repeating unit of X. laevis DNA. Cell. 1980 Feb;19(2):345–353. doi: 10.1016/0092-8674(80)90509-7. [DOI] [PubMed] [Google Scholar]
  17. Philipson L., Andersson P., Olshevsky U., Weinberg R., Baltimore D., Gesteland R. Translation of MuLV and MSV RNAs in nuclease-treated reticulocyte extracts: enhancement of the gag-pol polypeptide with yeast suppressor tRNA. Cell. 1978 Jan;13(1):189–199. doi: 10.1016/0092-8674(78)90149-6. [DOI] [PubMed] [Google Scholar]
  18. Rawlins D. R., Muzyczka N. Construction of a specific amber codon in the simian virus 40 T-antigen gene by site-directed mutagenesis. J Virol. 1980 Nov;36(2):611–616. doi: 10.1128/jvi.36.2.611-616.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Razin A., Hirose T., Itakura K., Riggs A. D. Efficient correction of a mutation by use of chemically synthesized DNA. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4268–4270. doi: 10.1073/pnas.75.9.4268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Silberklang M., Gillum A. M., RajBhandary U. L. Use of in vitro 32P labeling in the sequence analysis of nonradioactive tRNAs. Methods Enzymol. 1979;59:58–109. doi: 10.1016/0076-6879(79)59072-7. [DOI] [PubMed] [Google Scholar]
  21. Temple G. F., Dozy A. M., Roy K. L., Kan Y. W. Construction of a functional human suppressor tRNA gene: an approach to gene therapy for beta-thalassaemia. Nature. 1982 Apr 8;296(5857):537–540. doi: 10.1038/296537a0. [DOI] [PubMed] [Google Scholar]
  22. Weiner A. M. An abundant cytoplasmic 7S RNA is complementary to the dominant interspersed middle repetitive DNA sequence family in the human genome. Cell. 1980 Nov;22(1 Pt 1):209–218. doi: 10.1016/0092-8674(80)90169-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES