Di Simone, Christopher (1992) The membrane fusion activities of native and reconstituted mumps and sendai viruses. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechTHESIS:09132011-085913260
The structure and activities of whole and reconstituted Sendai and mumps viruses were examined using a number of physical and biological techniques: electron microscopy, photon correlation spectroscopy, perturbed angular correlation spectroscopy, gel electrophoresis, hemagglutination assays, plaque assays, fluorescence microscopy and fluorescent assays of membrane fusion. The fluorescent probe octadecyl rhodamine (RIB) was found to have a proximal transfer behavior which reduced usage of the probe as a membrane fusion indicator to short time periods. Simple bireactant and mass action kinetic models were sufficent to fit the data provided by the fluorescent assays on viral fusion with cell membranes. The reconstitution of mumps virus was optimized by using the detergent Triton X-IOO. Reconstituted virus envelopes which were the same size as whole virus and which were active in binding and fusing to cells were produced. The reconstituted Sendai and mumps virus accumulated in the reticuloendothelial system when injected into mice and hamsters, respectively. The kinetics of the membrane fusion activity of mumps with ghost erythrocytes and CV-I cells was measured and analysed. A general rate of reaction of 3 (±1) x 10^9 M^-l sec^-1 was found. The membrane fusion activity of reconstituted Sendai with HL60, U937, Cos, H9 and PBL cell lines was also measured and analyzed. The fusion activity was utilized to deliver plasmids for transfection to the interiors of the HL60 and Cos cells. Luciferase expression was found in the Cos cells but not in the HL60 cells. Loading of plasm ids into the vesicle interiors was enhanced by a factor of ten by complexing the DNA with the positive proteins polylysine, lysozyme and protamine.
|Item Type:||Thesis (Dissertation (Ph.D.))|
|Degree Grantor:||California Institute of Technology|
|Division:||Chemistry and Chemical Engineering|
|Thesis Availability:||Restricted to Caltech community only|
|Defense Date:||17 October 1991|
|Default Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Dan Anguka|
|Deposited On:||13 Sep 2011 17:45|
|Last Modified:||26 Dec 2012 04:38|
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