Download PDF by Masayori Inouye: Molecular Biology of RNA. New Perspectives

By Masayori Inouye

ISBN-10: 012372483X

ISBN-13: 9780123724830

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R. (1986a). Science 231, 470-475. Zaug, A. , and Cech, T. R. (1986b). Biochemistry 25, 4478-4482. Zaug, A. , Grabowski, P. , and Cech, T. R. (1983). Nature {London) 301, 578-583. Zaug, A. , Kent, J. , and Cech, T. R. (1984). Science 224, 574-578. Zaug, A. , Kent, J. , and Cech, T. R. (1985). Biochemistry 24, 6211-6218. Zaug, A. , Been, M. , and Cech, T. R. (1986). Nature {London) 324, 429-433. 4 Processing and Genetic Characterization of Self-Splicing RNAs of Bacteriophage T4 MARLENE BELFORT,* JOAN PEDERSEN-LANE,* KAREN EHRENMAN,*t DWIGHT H.

Synthetic oligodeoxynucleotides c o m p l e m e n t a r y to strategic regions of the primary transcript or R N A splice products w e r e used as p r o b e s (Fig. 5). T h e intron-specific p r o b e (probe 1) that hybridizes to premessage and excised intron gave a signal of roughly equal intensity with wild-type and m u t a n t R N A s . On the o t h e r h a n d , p r o b e s 2 and 3, specific to the 5' and 3' p r e m e s s a g e splice sites, respectively, gave m o r e intense signals with the m u t a n t R N A s than with R N A from the wild-type infection.

T h e facile m e t h o d s described for isolating and mapping td mutations u n d e r physiological conditions, as well as the assay developed to screen splicing-defective m u t a n t s , places us in a position to analyze exhaustively libraries of td m u t a t i o n s . This saturation mutagenesis a p p r o a c h should facilitate a division of d o m a i n s for splicing into functional subdomains and should eventually yield a complete genetically authenticated secondary structure model for the td r i b o z y m e .

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Molecular Biology of RNA. New Perspectives by Masayori Inouye


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