The current status of two-dimensional electrophoresis in germ cell mutation research
Previous research demonstrated that isoelectric focusing followed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis can be used to resolve hundreds of proteins from a single sample into a pattern of well-defined polypeptide spots. The possible application of this two-dimensional electrophoresis (2DE) technique to the detection of heritable mutations was recognized and demonstrated using mouse tissues. The studies demonstrated that actual implementation of 2DE for large genetic studies, however, required rigorous pattern reproducibility and methods of extracting numerical data from the patterns. The development of equipment for multiple parallel 2DE analyses and of computer software for the analysis of the resulting 2DE patterns during the 10 years since the first description of 2DE have provided the necessary tools for the application of 2DE to genetic studies. We are currently using inbred strains of mice to study the mutation detection capability of 2DE. The levels of sample and pattern reproducibility required for detection of significant changes in protein expression are being defined; the detection of mutations induced by different classes of mutagen (e.g., those causing large versus small alterations in DNA) is being assessed; and the population of proteins (and therefore genes) monitored by 2DE analysis is being characterized. 19 refs., 3 figs.
- Research Organization:
- Argonne National Lab., IL (USA)
- Sponsoring Organization:
- DOE/ER
- DOE Contract Number:
- W-31109-ENG-38
- OSTI ID:
- 5484419
- Report Number(s):
- CONF-8907113-13; ON: DE90002191
- Country of Publication:
- United States
- Language:
- English
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