A biotinylated MutS fusion protein and its use in a rapid mutation screening technique.
The Escherichia coli DNA mismatch repair protein, MutS, binds single base 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 biotinylated peptide domain has been constructed (MutSb). The biotinylation tag facilitates MutS detection and binding by avidin without significantly altering the DNA mismatch binding properties of MutS in vitro. We describe a novel and rapid mutation detection method with MutSb using streptavidin-coated magnetic beads and demonstrate that MutSb can also be used to remove mismatch containing DNA fragments from a mixture of DNA fragments in solution.
Geschwind
D H
DH
Department of Neurology, UCLA School of Medicine 90025, USA.
Rhee
R
R
Nelson
S F
SF
D000251:Adenosine Triphosphatases; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D001710:Biotin; D004260:DNA Repair; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D051719:MutS DNA Mismatch-Binding Protein; D015345:Oligonucleotide Probes; D017354:Point Mutation
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D015345:Oligonucleotide Probes; D001710:Biotin; D000251:Adenosine Triphosphatases; D051719:MutS DNA Mismatch-Binding Protein; C497175:MutS protein, E coli