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Published February 1, 1978 | public
Journal Article Open

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

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.

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August 22, 2023
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October 16, 2023