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Gene mapping and gene enrichment by the avidin-biotin interaction: use of cytochrome-c as a polyamine bridge

Sodja, Ann and Davidson, Norman (1978) Gene mapping and gene enrichment by the avidin-biotin interaction: use of cytochrome-c as a polyamine bridge. Nucleic Acids Research, 5 (2). pp. 385-401. ISSN 0305-1048. http://resolver.caltech.edu/CaltechAUTHORS:SODnar78

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Abstract

A modification of previously described methods of electron microscopic gene mapping and of gene enrichment based on the avidin-biotin interaction is presented. The modification consists of coupling cytochrome-c instead of pentane tane diamine to the oxidized 2', 3' terminus of an RNA by Schiff base formation and reduction. The RNA-cytochrome-c conjugate is purified by a simple chromatographic procedure; several biotins are attached to the cytochrome moiety by acylation. The extended arm between biotin and RNA gives efficient electron microscopic gene mapping of DNA:RNA-biotin hybrids with avidin-ferritin and avidin-polymethacylate sphere labels and efficient gene enrichment by buoyant banding of DNA:RNA-biotin:avidin-spheres in CsCl. A 1400 fold enrichment (thus, 25% pure) and a 90% yield of long Drosophila DNA strands with 5S RNA genes is achieved.


Item Type:Article
Additional Information:Copyright © 1978 Oxford University Press. Received October 31, 1977. We would like to thank Dr. Maria Pellegrini for helpful discussions on the enrichment experiments. A.S. was the recipient of a fellowship from the California Section of the American Cancer Society. This research has been supported by grant GM 10991 from the United States Public Health Service.
Record Number:CaltechAUTHORS:SODnar78
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:SODnar78
Alternative URL:http://nar.oxfordjournals.org/cgi/content/abstract/5/2/385
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4043
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:25 Jul 2006
Last Modified:26 Dec 2012 08:57

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