Sendal virus envelopes (SVE) were isolated from Sendal virus particles by Triton X-100 solubilization and ultracentrifugation. The envelopes were reconstituted in the presence of the fluorescent dye calcein by gradual removal of the detergent with Bio-beads SM-2. The internal volume of reconstituted Sendal virus envelopes (RSVE) was determined by quenching the fluorescence of calcein with cobalt (II) ions. The internal volume of RSVE was found to be proportional to the initial SVE protein concentration in the recon-stitution mixture, reaching about 18% of the total volume with 5.6 mg of SVE protein per ml. When radiolabelled cloned Epstein-Barr virus DNA fragment was included in the reconstitution mixture, the proportion of DNA associated with the vesicles much exceeded the trapping volume, indicating adsorption of DNA to the internal surface of RSVE. These deter-minations will allow optimization of the use of RSVE as gene-transfer vehicles.
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July 01 1984
Determination of the internal volume of reconstituted Sendai virus envelopes by quenching of calcein fluorescence
Ronald L. Bartzatt;
Ronald L. Bartzatt
1Department of Pathology and Laboratory Medicine and Eppley Institute for Research in Cancer, University of Nebraska Medical Center, Omaha, Nebraska 68105, USA
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David J. Volsky
David J. Volsky
1Department of Pathology and Laboratory Medicine and Eppley Institute for Research in Cancer, University of Nebraska Medical Center, Omaha, Nebraska 68105, USA
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Publisher: Portland Press Ltd
Received:
June 12 1984
Online ISSN: 1573-4935
Print ISSN: 0144-8463
© 1984 The Biochemical Society
1984
Biosci Rep (1984) 4 (7): 551–557.
Article history
Received:
June 12 1984
Citation
Ronald L. Bartzatt, David J. Volsky; Determination of the internal volume of reconstituted Sendai virus envelopes by quenching of calcein fluorescence. Biosci Rep 1 July 1984; 4 (7): 551–557. doi: https://doi.org/10.1007/BF01121911
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