Abstract
Interaction defective alleles (IDAs) are alleles that contain mutations affecting their ability tointeract with their wild type binding partners. The locations of the mutations may lead to the identificationof protein interaction domains and interaction interfaces. IDAs may also distinguish different bindinginterfaces of multidomain proteins that are part of large complexes, thus shedding light on large proteinstructures that have yet to be determined. IDAs may also be used in conjunction with RNAi to dissect proteininteraction networks. Here, the wild type allele is knocked down and replaced with an IDA that has lostthe ability to interact with a specific binding partner. As a result, interactions are disruptedrather than knocking out the entire gene. Thus, IDAs have the potential to be extremely valuable toolsin protein interaction network analysis. IDAs can be isolated by reverse two-hybrid analysis, which wasdemonstrated over a decade ago, but high background levels caused by truncated IDAs have preventedits widespread adoption. We recently described a novel method for full-length allele library generationthat eliminates this background and increases the efficiency of the reverse two-hybrid protocol (and IDAisolation) significantly. Here we discuss our strategy for allele library generation, the potential usesof IDAs as outlined above, and additional applications of allele libraries.
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Chappell, T.G., Gray, P.N. (2008). Protein Interactions: Analysis Using Allele Libraries. In: Werther, M., Seitz, H. (eds) Protein – Protein Interaction. Advances in Biochemical Engineering/Biotechnology, vol 110. Springer, Berlin, Heidelberg. https://doi.org/10.1007/10_2008_102
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DOI: https://doi.org/10.1007/10_2008_102
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