Investigation of Tandem Mass Spectrometry for Identifying Single and Mass-Compensating Double Base Substitutions in p53 and Bacterial Ribosomal PCR Products
EMSL Project ID
1791
Abstract
The p53 gene has been recognized in the past few years as an important cell cycle regulatory gene implicated in the production of cancers. One form of the altered gene implicated in cancers possesses a single base mutation. However, a common base substitution has also been identified within the p53 gene as a source of naturally occurring sequence variability within the population. Therefore, it is important to establish methods to examine forms of the gene that possess bases substitutions that may arise by both natural variability and induced mutations. The previous work by both groups indicates that mass spectrometry is capable of examining PCR products from each type of gene. The focus of this project is to investigate the use of tandem mass spectrometry to provide additional identifying information on sequence variations between PCR products. In situations where slight mass differences between relatively large products are difficult to identify, dissociation of the products into smaller fragment ions might make these differences more easily recognizable. A further aspect of this study is to examine products that have more than one base substitution, but have an identical mass as a result. This could happen if the second substitution is the opposite base switch which will result in each strand having identical base compositions, but different sequences. This situation will make it impossible to differentiate the products in the first stage of the mass spectrometer. Mass analysis after dissociation of each product should yield fragments of different masses allowing for differentiation of these products.
Project Details
Project type
Exploratory Research
Start Date
2000-01-06
End Date
2000-06-01
Status
Closed
Released Data Link
Team
Principal Investigator