Molecular Basis of Aging by A. Roy (Eds.)

By A. Roy (Eds.)

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T h u s , an i n c r e a s e o f e r r o r s in p r o t e i n synthesis during a g i n g has not been u n e q u i v o c a l l y e s t a b l i s h e d and e v e n when i n d i r e c t l y i n f e r r e d h a s b e e n l i m i t e d t o particular proteins. III. METHODS TO STUDY THE F I D E L I T Y OF DNA SYNTHESIS Two d i f f e r e n t a p p r o a c h e s h a v e b e e n d e v e l o p e d t o s t u d y t h e a c c u r a c y o f c a t a l y s i s b y DNA p o l y m e r a s e s . B o t h o f t h e s e h a v e b e e n u t i l i z e d i n s t u d i e s o n DNA polymerase from e u c a r y o t i c c e l l s .

Shmookler R e i s , R . J . A c a d . S e i . USA 7 9 , Macieira-Coelho, Α . , Exp. C e l l R e s . 4 2 , and Goldstein, and G o l d s t e i n , 3949 (1982). P o n t e n , J . and 673 (1966). Macieira-Coelho, Α . , Mech. , Cell Proc. 21, Natl. , D e v . C>, 341 (1977). Y a n i s h e v s k y , R . , M e n d e l s o h n , M . L . , M a y a l l , B . H . and C r i s t o f a l o , V . , J. C e l l P h y s i o l . 8 4 , 165 ( 1 9 7 4 ) . H a r l e y , C . B . , S h m o o k l e r R e i s , R . J . and G o l d s t e i n , S .

A. Use o f Polynucleotide Templates Polynucleotide templates offer a very simple t e c h n i q u e t o a s s a y t h e f i d e l i t y o f DNA s y n t h e s i s . In p r i n c i p l e one c o p i e s a t e m p l a t e such as p o l y d ( A - T ) , w h i c h c o n t a i n s o n l y r e s i d u e s o f d e o x y a d e n o s i n e and deoxythymidine in a l t e r n a t i n g s e q u e n c e s , and o n e measures the i n c o r p o r a t i o n o f e i t h e r complementary n u c l e o t i d e s u b s t r a t e , d ΑΤΡ o r d ΤΤΡ and a n o n - c o m p l e m e n t a r y s u b s t r a t e , e i t h e r dCTP o r d G T P .

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