Abstract
Cellular phenotypes resulting from decreased function of a specific gene are manifested as recessive, since they are suppressed in the presence of a normal allele of the corresponding gene. The powerful gene-transfer techniques, which have played a key role in the studies of dominant phenotypes, are not readily applicable to recessive traits, since expression of a recessive allele does not generally affect the cellular phenotype. In haploid organisms, random gene disruption can be used as a general method of cloning recessive genes, but this approach is limited to genes that are not essential for cell growth and is not, at least presently, applicable to diploid cells. This explains why studies of recessive genes of higher eukaryotes (e.g., tumor suppressor genes) are in many cases lagging far behind the analysis of dominant genes (e.g., oncogenes).
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References
Roninson, I. B., Gudkov, A. V., Holzmayer, T. A., Kirschling, D. J., Kazarov, A. R., Zelinck, C. R., Mazo, I. A., Axenovich, S. A., and Thimmapaya, R. (1995) Genetic suppressor elements: new tools for molecular oncology. Cancer Res. 55, 4023–4028.
Holzmayer, T. A., Pestov, D. G., and Roninson, I. B. (1992) Isolation of dominant negative mutants and inhibitory antisense RNA sequences by expression selection of random DNA fragments. Nuclerc Acids Res. 20, 711–717.
Gudkov, A. V., Zelnick, C., Kazarov, A. R., Thimmapaya, R., Suttle, D. P., Beck, W. T., and Roninson, I. B. (1993) Isolation of genetic suppressor elements, inducing resistance to topoisomerase II-interactive cytotoxic drugs, from human topoisomerase II cDNA. Proc. Natl. Acad. Sci. USA 90, 3231–3235.
Ossovskaya, V. S., Mazo, I. A., Strezoska, Z., Chernova, O. B., Chernov, M. V., Stark, G. R., Chumakov, P. M., and Gudkov, A. V. (1996) Use of genetic suppressor elements to dissect distinct biological effects of separate p53_domains. Proc. Natl. Acad. Sci. USA 93, in press.
Tarasewich, D. G., Schott, B., Xuan, Y., Chang, B., Salov, S., Holzmayer, T. A., and Roninson, I. B. (1996) Isolation of a genetic element inhibiting BCL-2 expression and sensitizing tumor cells to chemotherapeutic drugs. In preparation.
Pestov, D. G. and Lau, L. F. (1994) Genetic selection of growth inhibitory sequences in mammalian cells. Proc. Natl. Acad. Sci. USA 91, 12,549–12,553.
Gudkov, A. V., Kazarov A. R., Thimmapaya, R., Mazo, I. A., Axenovrch, S., and Roninson, I. B. (1994) Isolation of genetic suppressor elements from a retroviral normalized cDNA library: identification of a kinesin associated with drug sensitivity and senescence. Proc. Natl. Acad. Sci. USA 91, 3744–3748.
Miller, A. D. and Rosman, G. J. (1989) Improved retroviral vectors for gene transfer and expression. BioTechniques 7, 980–986.
Pear, W. S., Nolan, G. P., Scott, M. L., and Baltimore, D. (1993) Production of high titer helper-free retroviruses by transient transfection. Proc. Natl. Acad. Sci. USA 90, P8392–8396.
Pear, W. S., Scott, M. L., Baltimore, D., and Nolan, G. (1995) Production of high-titer helper-free retroviruses with an amphotropic host range by transient transfection. In preparation.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Patanjali, S. R., Parimoto, S., and Weissman, S. M. (1991) Construction of a uniform abundance (normalized) cDNA library. Proc. Natl. Acad. Sci. USA 88, 1943–1947.
Markowitz, D., Gaff, S., and Bank, A. (1988) Construction and use of a safe and efficient amphotropic packaging cell line. Virology 167, 400–406.
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© 1997 Humana Press Inc., Totowa, NJ
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Gudkov, A.V., Roninson, I.B. (1997). Isolation of Genetic Suppressor Elements (GSEs) from Random Fragment cDNA Libraries in Retroviral Vectors. In: Cowell, I.G., Austin, C.A. (eds) cDNA Library Protocols. Methods in Molecular Biology™, vol 69. Humana Press. https://doi.org/10.1385/0-89603-383-X:221
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DOI: https://doi.org/10.1385/0-89603-383-X:221
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