Transfection of a Temperature Sensitive V-MOS Oncogene in Chinese Hamster Ovary Cells
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Authors
Crumm, Emily
Issue Date
1988
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
The viral mos oncogene derived from the Moloney murine
sarcoma virus, is capable of forming cancerous tumors in suitable
animal tissues. The Chinese hamster ovary cell line has been used
to study v-mos protein kinase activity in vitro. One
significant effect of v-mos transfection into Chinese hamster
ovary cells is the increased presence of a 25 kilodalton
phosphoprotein. The object of this study was to use a temperature
sensitive cell line to induce the activity of the v-mos gene
and correlate the phosphorylation of the 25 kilodalton protein
with v-mos expression. A temperature sensitive v-mos gene
was used to study v-mos expression and the phosphorylation of
the 25 kilodalton phosphoprotein. By using a temperature
sensitive gene, the expression of the v-mos oncogene could be
suppressed and then reinstated. The temperature sensitive 159
neo (ts159neo) plasmid was transfected into both the wild
type and a mutant (10248) cell line deficient in cyclic AMP-dependent
protein kinase activity. The morphology, growth rates,
and 8-bromo-cyclic AMP sensitivity were determined for each of
the clones at both the permissive (32°C) and nonpermissive (39°C)
temperatures for v-mos expression. One and two dimensional
gel electrophoresis procedures were used to determine the
presence of the 25 kilodalton phosphoprotein at both
temperatures. One clone (10248 3D1) was found to be temperature
sensitive in the phosphorylation of the 25 kilodalton protein.
These results suggest that the v-mos oncogene may be acting
directly to phosphorylate the 25 kilodalton protein.
Description
vii, 45 p.
Citation
Publisher
Kalamazoo College
License
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