title: A method for the incremental expansion of polyglutamine repeats in recombinant proteins.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia.
authors.forename: Amy L
authors.initials: AL
authors.lastname: Robertson
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Bottomley
mesh_terms: D000818:Animals; D006801:Humans; D010455:Peptides / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D020449:Repetitive Sequences, Amino Acid; D019680:Trinucleotide Repeat Expansion
chemical_list: D010455:Peptides; D011994:Recombinant Proteins; C097188:polyglutamine