Abstract
THE availability of methods for the amplification of nucleic acid sequences1,2 allows the genetic analysis in vitro of the structural and functional properties of many nucleic acids. We have now developed an in vitro selection and amplification system, and used it to analyse the self-splicing Tetrahymena ribozyme. A much wider range of selective conditions can be used in vitro than is possible with standard in vivo methods, and many more variants can be handled in vitro than in vivo. This method can be used to isolate the wild-type ribozyme, and structural variants that are as active as the wild type, from a pool of over 250,000 variants in only three cycles of selection and amplification.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Saiki, R. K. et al. Science 239, 487–491 (1988).
Kwoh, D. Y. et al. Proc. natn. Acad. Sci. U.S.A. 86, 1173–1177 (1989).
Cech, T. R. & Bass, B. L. A. Rev. Biochem. 55, 599–630 (1986).
Davies, R. W., Waring, R. B., Ray, J. A., Brown, T. A. & Scazzochio, C. Nature 300, 719–724 (1982).
Couture, S. et al. J. molec. Biol. (in the press).
Kim, S.-H. & Cech, T. R. Proc. natn. Acad Sci. U.S.A 84, 8788–8792 (1987).
Michel, F. & Westhof, E. J. molec. Biol. (in the press).
Robertson, D. L. & Joyce, G. F. Nature 344, 467–468 (1990).
Burke, J. M. et al. Nucleic Acids Res. 15, 7217–7221 (1987).
Milligan, J. F., Groebe, D. R., Witherell, G. W. & Uhlenbeck, O. C. Nucleic Acids Res. 15, 8783–8798 (1987).
Doudna, J. A., Gerber, A. S., Cherry, J. M. & Szostak, J. W. Cold Spring Harb. Symp. quant. Biol. 52, 173–180 (1986).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Green, R., Ellington, A. & Szostak, J. In vitro genetic analysis of the Tetrahymena self-splicing intron. Nature 347, 406–408 (1990). https://doi.org/10.1038/347406a0
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1038/347406a0
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.