Harrington, Michael G. and Coffman, James A. and Calzone, Frank J. and Hood, Leroy E. and Britten, Roy J. and Davidson, Eric H. (1992) Complexity of Sea Urchin Embryo Nuclear Proteins That Contain Basic Domains. Proceedings of the National Academy of Sciences of the United States of America, 89 (14). pp. 6252-6256. ISSN 0027-8424 http://resolver.caltech.edu/CaltechAUTHORS:HARpnas92
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We describe a quantitative two-dimensional gel electrophoretic analysis of nuclear extract from 24-hr sea urchin embryos. The extract was fractionated by using a weak cation-exchange resin, and eight known DNA-binding proteins were shown to be entirely included in a salt eluate that releases proteins containing basic domains. This fraction and a lower-salt fraction containing the majority of the protein species were mapped two-dimensionally by using new algorithms that permit reproducible spot identification, storage of intensity and map-position data, and subtractive comparison of one pattern with respect to another. By reference to a previously characterized DNA-binding factor, spot intensity could be interpreted in terms of the number of molecules per embryo nucleus. A map was constructed displaying all nuclear proteins containing basic domains that are present within the concentration range per nucleus of a set of known DNA-binding factors of the sea urchin embryo. The map includes 265 spots that fulfill both of these criteria, probably representing about 100 different protein species.
|Additional Information:||Copyright © 1992 by National Academy of Sciences Contributed by Eric H. Davidson, April 6, 1992 All 2-D electrophoretic gels were skillfully and reproducibly run by Miki Yun, to whom we are very grateful. We also thank Robert W. Zeller for his generous donation of the P3A1- and P3A2-specific antisera, and Wei Wu for help with computer programming. J.A.C. was supported by National Institutes of Health Training Grant HD-07257. This work was supported by National Science Foundation Grant STCDIR 8809719 (to L.E.H. and M.G.H.), a grant from the Ralph M. Parsons Foundation (to L.E.H. and E.H.D.), National Institutes of Health Grant HD05753 (to E.H.D.) and National Science Foundation Grant DCB8912530 (to F.J.C.). 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:||nuclear extract; cation-exchange chromatography; image analysis; two-dimensional mapping|
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|Deposited By:||Archive Administrator|
|Deposited On:||05 Sep 2006|
|Last Modified:||26 Dec 2012 09:01|
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