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Intracellular centrifugal separation of organelles in Phycomyces

Zalokar, Marko (1969) Intracellular centrifugal separation of organelles in Phycomyces. Journal of Cell Biology, 41 (2). pp. 494-509. ISSN 0021-9525. http://resolver.caltech.edu/CaltechAUTHORS:ZALjcb69

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Abstract

Live sporangiophores of Phycomyces blakesleeanus were centrifuged at 35,000 rpm. The cell contents sedimented into distinct layers, and each layer was studied with an electron microscope and with cytochemical methods. The following layers were found (their volumes and their densities are shown in Fig. 3): 1. polyphosphates; 2. polyphosphates and protein crystals; 3. glycogen; 4. yellow layer with ferritin; 5. ribosomes; 6. protein crystals; 7. mitochondria; 8. mitochondria and fibrils; 9. nuclei; 10. endoplasmic reticulum; 11. vesicles, membranes, and reticulum; 12. vacuole; 13. lipoproteins, membranes; 14. fat droplet. The densities of the various layers were determined by the injection of droplets of inert oils of known density into the sporangiosphores before centrifugation. Sedimented cell organelles could be isolated. Centrifuged nuclei of a lycopene-producing mutant were injected into the intact sporangiophore of an albino host where they induced color formation. The ensuing spores, when plated, gave a mixture of white and colored colonies. It was concluded that cell organelles, sedimented by centrifugation of living sporangiophores, remain alive and can be used for biochemical studies. Microspectrophotometric examination of the layers indicated the presence of cytochromes and flavines in the mitochondria and of cytochromes in the nuclei. No pigments corresponding to the action spectrum for the light growth response were found.


Item Type:Article
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http://jcb.rupress.org/cgi/content/abstract/41/2/494PublisherUNSPECIFIED
Additional Information:© 1969 Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. Received for publication 15 October 1968, and in revised form 30 December 1968. This work was supported by the Gosney Fund and by a National Science Foundation grant GB-4642. The author wishes to thank Dr. M. Delbrück for helpful suggestions and criticism, Mrs. P.V. Burke for help in the preparation of the manuscript, and Dr. G. Meissner for the measurements of light absorption. G. Adam did some early work on centrifugation.
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Funding AgencyGrant Number
Gosney Fund, CaltechUNSPECIFIED
National Science FoundationGB-4642
Record Number:CaltechAUTHORS:ZALjcb69
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:ZALjcb69
Usage Policy:RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.
ID Code:12120
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 Nov 2008 01:40
Last Modified:26 Dec 2012 10:27

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