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          Institute: MPI für molekulare Physiologie     Collection: Sonstige wissenschaftliche Organisationseinheiten     Display Documents



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ID: 6185.0, MPI für molekulare Physiologie / Sonstige wissenschaftliche Organisationseinheiten
A non-radioactive protein truncation test for the sensitive detection of all stop and frameshift mutations
Authors:Kahmann, Sabine; Herter, Peter; Kuhnen, Cornelius; Müller, Klaus-Michael; Muhr, Gert; Martin, Dirk; Soddemann, Matthias; Müller, Oliver
Language:English
Date of Publication (YYYY-MM-DD):2002
Title of Journal:Human Mutation
Journal Abbrev.:Hum. Mutat.
Volume:19
Issue / Number:2
Start Page:165
End Page:172
Sequence Number of Article:1
Review Status:Peer-review
Audience:Experts Only
Intended Educational Use:No
Abstract / Description:A new method for mutation detection is described, which is a technical advancement of the protein truncation test. The new technique is non-radioactive and highly sensitive for detection of virtually all sequence mutations, which lead to a stop signal or to the shift of the translation frame. The method includes four steps: 1) capture of the interesting sequence copies out of the sample by binding to an immobilized complementary sequence, 2) PCR amplification of the gene fragment to be analyzed with primers coding both for amino. and carboxy-terminal tags, 3) in vitro transcription and translation, and 4) analysis of the translation products by Western blot. As an evaluation of the new method, we detected mutated gene copies at a dilution of 1 to 40 compared to the non,mutated gene. Using the method, we were able to detect a mutation in the adenomatous polyposis coli tumor suppressor gene (APC) in a stool sample of a colorectal cancer patient. This mutation could not be detected by direct sequencing of the amplified APC gene fragme
Free Keywords:adenomatous polyposis coli; APC; hybrid capture; in vitro synthesis protein assay; IVSP; mutation detection; protein truncation test; PTT; tumor suppressor
External Publication Status:published
Document Type:Article
Communicated by:Jürgen Block
Affiliations:MPI für molekulare Physiologie/Sonstige Wissenschaftliche Organisationseinheiten/AG Dr. Oliver Müller
External Affiliations:Ruhr-Universität Bochum, Berufsgenossenschaftliche Kliniken Bergmannsheil, Institut für Pathologie, D-4630 Bochum, Germany;
Ruhr-Universität Bochum, Berufsgenossenschaftliche Kliniken Bergmannsheil, Chirurgische Klinik, D-4630 Bochum, Germany
Identifiers:URL:http://www3.interscience.wiley.com/cgi-bin/fulltex... [pdf version of this article]
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