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. 1984 Apr;4(4):749–754. doi: 10.1128/mcb.4.4.749

Expression of complete chicken thymidine kinase gene inserted in a retrovirus vector.

P K Bandyopadhyay, H M Temin
PMCID: PMC368793  PMID: 6325895

Abstract

The chicken thymidine kinase (tk) gene was inserted into spleen necrosis virus. Thymidine kinase activity was expressed even when the promoter and terminator sequences for tk RNA synthesis were retained. When the promoter was present in the same orientation as the promoter in the long terminal repeat of the virus, deletions occurred both in the virus and in the tk gene, and the thymidine kinase-transforming activity of the recovered virus was low. Splicing of apparent intervening sequences in the tk gene was also observed. When the orientation of the tk promoter was opposite to the promoter in the long terminal repeat, virus synthesis was diminished, whereas thymidine kinase activity was expressed at an elevated level compared with virus in which the promoter was in the same orientation. However, when the apparent tk promoter was deleted from virus with the tk gene in the opposite orientation, a high level of virus synthesis was observed, probably as a result of absence of interference of RNA synthesis from converging promoters. The intervening sequences in the virus in which the promoters were in opposite orientation were not spliced.

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Selected References

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

  1. Emerman M., Temin H. M. High-frequency deletion in recovered retrovirus vectors containing exogenous DNA with promoters. J Virol. 1984 Apr;50(1):42–49. doi: 10.1128/jvi.50.1.42-49.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Hamer D. H., Leder P. Splicing and the formation of stable RNA. Cell. 1979 Dec;18(4):1299–1302. doi: 10.1016/0092-8674(79)90240-x. [DOI] [PubMed] [Google Scholar]
  3. Hamer D. H., Smith K. D., Boyer S. H., Leder P. SV40 recombinants carrying rabbit beta-globin gene coding sequences. Cell. 1979 Jul;17(3):725–735. doi: 10.1016/0092-8674(79)90279-4. [DOI] [PubMed] [Google Scholar]
  4. Kwoh T. J., Zipser D., Wigler M. Mutational analysis of the cloned chicken thymidine kinase gene. J Mol Appl Genet. 1983;2(2):191–200. [PubMed] [Google Scholar]
  5. Perkins A. S., Kirschmeier P. T., Gattoni-Celli S., Weinstein I. B. Design of a retrovirus-derived vector for expression and transduction of exogenous genes in mammalian cells. Mol Cell Biol. 1983 Jun;3(6):1123–1132. doi: 10.1128/mcb.3.6.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Perucho M., Hanahan D., Lipsich L., Wigler M. Isolation of the chicken thymidine kinase gene by plasmid rescue. Nature. 1980 May 22;285(5762):207–210. doi: 10.1038/285207a0. [DOI] [PubMed] [Google Scholar]
  7. Santangelo G. M., Cole C. N. Preparation of a "functional library" of African green monkey DNA fragments which substitute for the processing/polyadenylation signal in the herpes simplex virus type 1 thymidine kinase gene. Mol Cell Biol. 1983 Apr;3(4):643–653. doi: 10.1128/mcb.3.4.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Shimotohno K., Temin H. M. Formation of infectious progeny virus after insertion of herpes simplex thymidine kinase gene into DNA of an avian retrovirus. Cell. 1981 Oct;26(1 Pt 1):67–77. doi: 10.1016/0092-8674(81)90034-9. [DOI] [PubMed] [Google Scholar]
  9. Shimotohno K., Temin H. M. Loss of intervening sequences in genomic mouse alpha-globin DNA inserted in an infectious retrovirus vector. Nature. 1982 Sep 16;299(5880):265–268. doi: 10.1038/299265a0. [DOI] [PubMed] [Google Scholar]
  10. Sorge J., Hughes S. H. Splicing of intervening sequences introduced into an infectious retroviral vector. J Mol Appl Genet. 1982;1(6):547–559. [PubMed] [Google Scholar]
  11. Spandidos D. A., Wilkie N. M. Host-specificities of papillomavirus, Moloney murine sarcoma virus and simian virus 40 enhancer sequences. EMBO J. 1983;2(7):1193–1199. doi: 10.1002/j.1460-2075.1983.tb01566.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Tabin C. J., Hoffmann J. W., Goff S. P., Weinberg R. A. Adaptation of a retrovirus as a eucaryotic vector transmitting the herpes simplex virus thymidine kinase gene. Mol Cell Biol. 1982 Apr;2(4):426–436. doi: 10.1128/mcb.2.4.426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ward D. F., Murray N. E. Convergent transcription in bacteriophage lambda: interference with gene expression. J Mol Biol. 1979 Sep 15;133(2):249–266. doi: 10.1016/0022-2836(79)90533-3. [DOI] [PubMed] [Google Scholar]
  14. Wei C. M., Gibson M., Spear P. G., Scolnick E. M. Construction and isolation of a transmissible retrovirus containing the src gene of Harvey murine sarcoma virus and the thymidine kinase gene of herpes simplex virus type 1. J Virol. 1981 Sep;39(3):935–944. doi: 10.1128/jvi.39.3.935-944.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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