Geschwind D H, Rhee R, Nelson S F
Department of Neurology, UCLA School of Medicine 90025, USA.
Genet Anal. 1996 Oct;13(4):105-11. doi: 10.1016/s1050-3862(95)00160-3.
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.
大肠杆菌DNA错配修复蛋白MutS在体内和体外均可结合单碱基对错配和短缺失。为使该蛋白适用于突变检测，构建了大肠杆菌MutS与生物素化肽结构域的融合蛋白（MutSb）。生物素化标签便于抗生物素蛋白检测和结合MutS，且在体外不会显著改变MutS的DNA错配结合特性。我们描述了一种使用链霉抗生物素蛋白包被磁珠的、基于MutSb的新型快速突变检测方法，并证明MutSb还可用于从溶液中的DNA片段混合物中去除含错配的DNA片段。
