A method for the incremental expansion of polyglutamine repeats in recombinant proteins
Amy L Robertson1, Stephen P Bottomley
1Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia.
Abstract:
The polyglutamine diseases are caused by the expansion of CAG repeats. A key step in understanding the disease mechanisms, at the DNA and protein level, is the ability to produce recombinant proteins with specific length glutamine tracts which is a time-consuming first step in setting up in vitro systems to study the effects of polyglutamine expansion. Described here is a PCR-based method for the amplification of CAG repeats, which we used to incrementally extend CAG length by 3-5 repeats per cycle. This method could be translated into various contexts where amplification of repeating elements is necessary.
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