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Messenger RNA prevalence in sea urchin embryos measured with cloned cDNAs

Lasky, Laurence A. and Lev, Ze'ev and Xin, Ji-Hou and Britten, Roy J. and Davidson, Eric H. (1980) Messenger RNA prevalence in sea urchin embryos measured with cloned cDNAs. Proceedings of the National Academy of Sciences of the United States of America, 77 (9). pp. 5317-5321. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:LASpnas80

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Abstract

mRNA prevalence during sea urchin development was measured by treating cDNA clone colonies with labeled cDNAs transcribed from unfertilized egg and embryo poly(A)-RNAs. The number of cytoplasmic transcripts per embryo complementary to several clones was determined independently by titration with poly(A)-RNA in solution, and the amount of cDNA bound to these clones in colony hybridizations was shown to be proportional to the concentration of the respective poly(A)-RNAs in the embryo cytoplasm. At the gastrula stage, the most prevalent mRNA species occur in about 106 molecules per embryo. If all cells were equivalent, this would be a few hundred molecules per cell. By pluteus stage, the prevalence of some sequences has increased more than 10-fold. Most, though not all, sequences prevalent in later embryos are also present in the maternal RNA of the unfertilized egg. For most poly(A)-RNA sequences, the prevalence levels determined during oogenesis are maintained through the pluteus stage, whereas a minority of sequences display sharp stage-specific changes in representation during development.


Item Type:Article
Additional Information:© 1980 by the National Academy of Sciences. Contributed by Roy J. Britten, June 13, 1980. Reverse transcriptase was kindly provided by Dr. J.W. Beard at Life Sciences (under a contract from the National Cancer Institute). We thank Drs. Norman Davidson and Elliot Meyerowitz for their helpful and critical reviews of the manuscript. This research was supported by National Institutes of Health Grant HD-05753. L.A.L. was supported by National Institutes of Health Fellowship GM-06717, Z.L. by a Chaim Weizmann Fellowship and by a Gosney Fellowship, and J.-H.X. by the Shanghai Institute of Biochemistry and by the Church Fund of the Division of Biology. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:RNA titration; cDNA clone library; colony hybridization
Record Number:CaltechAUTHORS:LASpnas80
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:LASpnas80
Alternative URL:http://www.pnas.org/cgi/content/abstract/77/9/5317
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:9185
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:19 Nov 2007
Last Modified:14 Nov 2014 19:20

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