Gene mapping and gene enrichment by the avidin-biotin interaction: use of cytochrome-c as a polyamine bridge
- Creators
- Sodja, Ann
- Davidson, Norman
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.
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.Files
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Additional details
- Eprint ID
- 4043
- Resolver ID
- CaltechAUTHORS:SODnar78
- Created
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2006-07-25Created from EPrint's datestamp field
- Updated
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2019-10-02Created from EPrint's last_modified field