Suitability of two-dimensional electrophoretic protein separations for quantitative detection of mutations
Separation of proteins by two-dimensional electrophoresis (2DE) provides a powerful method for mutagenesis studies, since hundreds of proteins can be monitored simultaneously. In previous mutation studies in which 2DE has been used, only qualitative protein differences were monitored; quantitative protein variations were not evaluated. Although significant differences in protein abundance can be detected by eye, the large number of protein spots present in 2DE patterns together with the large number of individual patterns required for a mutagenesis study would necessitate the use of a computerized analysis system to detect the rare quantitative protein changes indicative of gene deletions or inactivation of genes by point mutations in regulatory genes. A pilot study to search for heritable mutations induced by treatment of mice with either ethylnitrosourea or gamma radiation is underway. Samples are being monitored for quantitative changes that reduce the amount of protein by about 50%. The results of this study indicate that the key methods to improve the application of 2DE to mutation screening are to increase the number of measurable spots (i.e., improve stain sensitivity) and to decrease the spread of values for the volume measurements. Even small improvements in these areas could greatly increase the number of monitorable spots. 9 refs., 4 figs.
- Research Organization:
- Argonne National Lab., IL (USA); Proteus Technologies, Inc., Rockville, MD (USA)
- DOE Contract Number:
- W-31109-ENG-38
- OSTI ID:
- 6847198
- Report Number(s):
- CONF-8609239-4; ON: DE87004757
- Country of Publication:
- United States
- Language:
- English
Similar Records
Two-dimensional gel electrophoresis for the detection of heritable mutations: Normal quantitive variability related to age
The current status of two-dimensional electrophoresis in germ cell mutation research
Related Subjects
560120 -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
560152 -- Radiation Effects on Animals-- Animals
560300 -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMALS
ELECTROMAGNETIC RADIATION
ELECTROPHORESIS
GAMMA RADIATION
GENE MUTATIONS
IMAGE PROCESSING
IONIZING RADIATIONS
MAMMALS
MICE
MUTATIONS
NITROSO COMPOUNDS
NITROSOUREAS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PROCESSING
PROTEINS
RADIATIONS
RODENTS
SCREENING
VERTEBRATES