de.mpg.escidoc.pubman.appbase.FacesBean
English
 
Help Guide Disclaimer Contact us Login
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

A non-radioactive protein truncation test for the sensitive detection of all stop and frameshift mutations

MPS-Authors

Kahmann,  Sabine
Max Planck Institute of Molecular Physiology, Max Planck Society;

Herter,  Peter
Max Planck Institute of Molecular Physiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons98712

Müller,  Oliver
Sonstige Wissenschaftliche Organisationseinheiten, Max Planck Institute of Molecular Physiology, Max Planck Society;

Locator
There are no locators available
Fulltext (public)
There are no public fulltexts available
Supplementary Material (public)
There is no public supplementary material available
Citation

Kahmann, S., Herter, P., Kuhnen, C., Müller, K.-M., Muhr, G., Martin, D., et al. (2002). A non-radioactive protein truncation test for the sensitive detection of all stop and frameshift mutations. Human Mutation, 19(2): 1, pp. 165-172. Retrieved from http://www3.interscience.wiley.com/cgi-bin/fulltext?ID=89012847&PLACEBO=IE.pdf.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0014-0F1B-A
Abstract
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