A BIOTINYLATED MUTS FUSION PROTEIN AND ITS USE IN A RAPID MUTATION SCREENING TECHNIQUE

The Escherichia coli DNA mismatch repair protein, MutS, binds single b ase pair mismatches and short deletions in vivo and in vitro. To adapt this protein for mutation detection, a fusion protein of E. coli MutS with a biotinylaled peptide domain has been constructed (MutSb). The biotinylation lag facilitates MutS detection and binding by avidin wit hout significantly altering the DNA mismatch binding properties of Mut S in vitro. We describe a novel and rapid mutation detection method wi th MutSb using streptavidin-coated magnetic beads and demonstrate that MutSb can also be used to remove mismatch containing DNA fragments fr om a mixture of DNA fragments in solution.