Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Oct;70(10):6810–6815. doi: 10.1128/jvi.70.10.6810-6815.1996

A replication-competent promoter-trap retrovirus.

J Coulombe 1, Y Avis 1, D A Gray 1
PMCID: PMC190726  PMID: 8794320

Abstract

A promoter-trap retrovirus has been constructed in which a promoterless polyomavirus middle T antigen gene was inserted in the U3 region of the long terminal repeat of a replication-competent Moloney murine leukemia virus. The resulting virus, designated PyT, was used to infect mouse mammary glands in situ. As expected, mammary tumors appeared in some infected animals. These tumors were found to contain PyT proviruses of the predicted structure. From one such tumor, the PyT provirus and surrounding sequences from the integration site were cloned. The provirus was found to have integrated adjacent to the promoter of a novel mouse gene (TRAP1) that was expressed at low levels in various mouse tissues. These data show that the PyT retrovirus provides a sensitive means of detecting active promoters in vivo.

Full Text

The Full Text of this article is available as a PDF (350.7 KB).

Selected References

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

  1. Adams M. D., Dubnick M., Kerlavage A. R., Moreno R., Kelley J. M., Utterback T. R., Nagle J. W., Fields C., Venter J. C. Sequence identification of 2,375 human brain genes. Nature. 1992 Feb 13;355(6361):632–634. doi: 10.1038/355632a0. [DOI] [PubMed] [Google Scholar]
  2. Chang W., Hubbard S. C., Friedel C., Ruley H. E. Enrichment of insertional mutants following retrovirus gene trap selection. Virology. 1993 Apr;193(2):737–747. doi: 10.1006/viro.1993.1182. [DOI] [PubMed] [Google Scholar]
  3. Friedrich G., Soriano P. Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes Dev. 1991 Sep;5(9):1513–1523. doi: 10.1101/gad.5.9.1513. [DOI] [PubMed] [Google Scholar]
  4. Gossler A., Joyner A. L., Rossant J., Skarnes W. C. Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes. Science. 1989 Apr 28;244(4903):463–465. doi: 10.1126/science.2497519. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Guy C. T., Cardiff R. D., Muller W. J. Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol Cell Biol. 1992 Mar;12(3):954–961. doi: 10.1128/mcb.12.3.954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. McBurney M. W., Sutherland L. C., Adra C. N., Leclair B., Rudnicki M. A., Jardine K. The mouse Pgk-1 gene promoter contains an upstream activator sequence. Nucleic Acids Res. 1991 Oct 25;19(20):5755–5761. doi: 10.1093/nar/19.20.5755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Reik W., Weiher H., Jaenisch R. Replication-competent Moloney murine leukemia virus carrying a bacterial suppressor tRNA gene: selective cloning of proviral and flanking host sequences. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1141–1145. doi: 10.1073/pnas.82.4.1141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Rohdewohld H., Weiher H., Reik W., Jaenisch R., Breindl M. Retrovirus integration and chromatin structure: Moloney murine leukemia proviral integration sites map near DNase I-hypersensitive sites. J Virol. 1987 Feb;61(2):336–343. doi: 10.1128/jvi.61.2.336-343.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Rowe W. P., Pugh W. E., Hartley J. W. Plaque assay techniques for murine leukemia viruses. Virology. 1970 Dec;42(4):1136–1139. doi: 10.1016/0042-6822(70)90362-4. [DOI] [PubMed] [Google Scholar]
  11. Skarnes W. C., Auerbach B. A., Joyner A. L. A gene trap approach in mouse embryonic stem cells: the lacZ reported is activated by splicing, reflects endogenous gene expression, and is mutagenic in mice. Genes Dev. 1992 Jun;6(6):903–918. doi: 10.1101/gad.6.6.903. [DOI] [PubMed] [Google Scholar]
  12. Vijaya S., Steffen D. L., Robinson H. L. Acceptor sites for retroviral integrations map near DNase I-hypersensitive sites in chromatin. J Virol. 1986 Nov;60(2):683–692. doi: 10.1128/jvi.60.2.683-692.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wang B. C., Kennan W. S., Yasukawa-Barnes J., Lindstrom M. J., Gould M. N. Carcinoma induction following direct in situ transfer of v-Ha-ras into rat mammary epithelial cells using replication-defective retrovirus vectors. Cancer Res. 1991 May 15;51(10):2642–2648. [PubMed] [Google Scholar]
  14. von Melchner H., DeGregori J. V., Rayburn H., Reddy S., Friedel C., Ruley H. E. Selective disruption of genes expressed in totipotent embryonal stem cells. Genes Dev. 1992 Jun;6(6):919–927. doi: 10.1101/gad.6.6.919. [DOI] [PubMed] [Google Scholar]
  15. von Melchner H., Ruley H. E. Identification of cellular promoters by using a retrovirus promoter trap. J Virol. 1989 Aug;63(8):3227–3233. doi: 10.1128/jvi.63.8.3227-3233.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES