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Plant Physiology 47:499-503 (1971)
© 1971 American Society of Plant Biologists

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Mass Isolation of Pea Nuclei

Kestutis J. Tautvydas1,2

a Department of Biology, Yale University, New Haven, Connecticut 06520

Improvements in conventional filtration and centrifugation procedures made it possible to increase the yield of intact cytoplasm-free pea (Pisum sativum) nuclei from the usual 3 to 10% to 32% (6 x 108 nuclei per 7 grams fresh weight of pea apices) and to complete the isolation in 80 to 90 minutes. The isolated nuclei appeared to retain their structural integrity as revealed in electron photomicrographs, and remained intact for at least 5 hours at 20 Celsius.

The DNA:RNA: protein ratio of isolated pea nuclei was found to be 3.1:1:9.9. Their RNA polymerase activity, monitored by incorporation of 14C into RNA from 14C-UTP, was linear for about 10 minutes, and then gradually declined over the next 15 to 20 minutes.

1 The author was aided by a Title IV NDEA fellowship. This represents a portion of a thesis submitted to the Graduate School at Yale University in 1969, in partial fulfillment of the requirements for a Ph.D. degree.

2 Present address: Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Colorado 80302.






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Copyright © 1971 by the American Society of Plant Biologists