[Comparative study on protein components of ouabain resistant mutant and wild cells of Chinese hamster V79 cell line by two-dimensional gel electrophoresis].
In the sense of gene expression, the primary and direct change in mutant cells should be involved in the protein component. Two-dimensional polyacrylamide gel electrophoresis has been used to compare the protein components of ouabain resistant mutant cells (ouaR) and sensitive wild type cells (ouaS) of chinese hamster lung cell line V79. The procedure is based on O'Farrell's method and we have made some modifications. In order to make the study reliable, we took twenty polypeptide spots as a reference marker. These marker spots could always be reproduced in a series of parallel experiments with both mutant and wild cells. In addition, these markers covered different small areas, representing different isoelectric point (pI) and molecular weight (MW) on the electrophoretogram. With reference to these marker spots, some polypeptide spots showed variations in their position, size and density. It is notable that the changes appeared in the small area 4 (pI/MW:5.0/28-30kd) and the small area (pI/MW:6.3/39kd) is highly reproducible and significant, so that these changes might be specific to ouabain resistant mutant of V79 cells. Since the level of genetic polymorphism of mammalian cells on two-dimensional gel electrophoresis is quite low, these specific changes might reflect the differences in gene structure and expression. This primary study should enhance the usefulness of two-dimensional gel electrophoresis for mutagenesis research with cultured mammalian somatic cells.
Zhou
X L
XL
Fu
J L
JL
Qi
J
J
Li
D C
DC
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004351:Drug Resistance / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D005819:Genetic Markers / Q000032:analysis*; D008168:Lung / Q000166:cytology*; D009154:Mutation; D010042:Ouabain; D010455:Peptides / Q000032:analysis*; D011110:Polymorphism, Genetic
D005819:Genetic Markers; D010455:Peptides; D010042:Ouabain