Isolation of Temperature Sensitive Mutants of Tanapox virus
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Authors
Grinfeld, Sonja
Issue Date
2009
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Tanapox virus (TPV) is a member of the Poxviridae family. Because it
causes transient limiting disease, the biogenesis of TPV is unclear. Use of
temperature sensitive (ts mutants), unknown gene functions and the biogenesis
can be determined. Such knowledge would be useful in engineering a safe and
effective TPV oncolytic virus. In addition, ts mutants might be able to serve as
an additional mechanism in case an oncolytic virus mutated again. In this
experiment, our objective is to isolate a number of temperature sensitive (ts)
mutants with a permissive temperature of 32.5 °C and a restrictive temperature of
37°C using nitrosoguanidine (N-G) as a mutagen. We used 100X-concentrated
TPV-GFP with N-G and diluted it 10-fold. Infections of TPV-GFP in OMK cells
were effective, and dilutions of10-7 from 100X concentration of mutated TPV-GFP
produced the ideal amount of isolated plaques. None of the 108 plaques
was ts, and the majority of the first 23 plaques grew better at the restrictive
temperature. More plaques need to be isolated to obtain a pool of ts TPV
mutants. Once obtained, these mutants would be useful in upcoming
experiments, such as creating a second block for a TPV oncolytic virus and
determining the biogenesis of TPV.
Description
vi, 20 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.