Incorporation of tritiated cytidine into ribonucleic acid by isolated pea nuclei

Creators: Rho, Joon H.; Chipchase, Margaret I.

Abstract

A new method for the isolation of the nuclei from plant tissue has been devised. This method consists in passing the plant tissue through a set of spring-loaded, counter-rotating rollers and collecting the liberated nuclei in sucrose solution containing calcium ions. A semi-automatic machine which couples the rollers with a special tissue chopping device has been constructed. It has been shown that nuclei obtained in this way actively incorporate cytidine-H3 into RNA. This incorporation is increased in the presence of nucleoside triphosphates and an energy-regenerating system.

Additional Information

© 1962 by Rockefeller University Press Received for publication, February 1, 1962. Report of work supported in part by Grant No. RG-5143, National Institutes of Health, United States Public Health Service, and Grant No. A-3102, United States Public Health Service. Dr. Rho is supported by the Visiting Scientists Program, National Academy of Sciences. We wish to thank Professor James Bonner and Dr. Max Birnstiel for their valuable advice and interest in this work, and Mr. Robert Hayes and Mr. Frank Ostrander for their technical assistance. The authors are indebted to Mr. Frank Ostrander, instrument maker, Division of Biology, California Institute of Technology, for the development and construction of this semi-automatic machine, and to Dr. Clifford Sato, for his original suggestion of the squeezing method.

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