Dr. Kornberg used his in vitro system to study the properties of DNA replication.  The final proof of the Watson and Crick Model was the Nearest Neighbor model.

In vitro replication of DNA using alpha P label- Arthur Kornberg

dAPPP +
dCPPP +
dGPPP + Primer + Template --> DNA + nPP's
(d)TPPP +

Digestion of DNA Bovine Spline Phosphodiesterase  (BSP) and Snake Venome Phosphodiesterase (SnVP)

• with BSP yields a 3'dAMP, 3'dCMP, 3' dGMP, and 3' TMP
• with SnVP yields a 5'dAMP, 5'dCMP, 5' dGMP, and 5' TMP

How can you get 3'monophosphates?

Kornberg added radioactive isotopes one at a time and measured the products after digestion with Bovine Spline Phosphodiesterase  (BSP) and Snake Venome Phosphodiesterase (SnVP). He found in every case that the results were compatable with the W&C Model for DNA ie the strands were antiparallel.

Text iGenetics by Peter J. Russell

This web site is provided for instruction in Botany and Zoology 342

by Kenneth G. Wilson,
Professor of Botany
Miami University
wilsonkg@muohio.edu