Merlo D J, Thompson D V
Anal Biochem. 1987 May 15;163(1):79-87. doi: 10.1016/0003-2697(87)90095-9.
Sodium bisulfite treatment of single-stranded DNA deaminates exposed cytosine residues to form uracil, resulting in cytosine-to-thymidine transition mutations following DNA replication. We have used this reaction in vitro to destroy the recognition sequences for the restriction endonucleases HindIII and XmaI in the aminoglycoside 3'-phosphotransferase I coding region of plasmid pUC4K. This procedure should be applicable to the mutation of any recognition sequence of restriction endonucleases which generate cytosine-containing single-stranded ends. The possibility of mutagenesis of restriction sites to generate stop codons in coding regions is discussed.
亚硫酸氢钠处理单链DNA会使暴露的胞嘧啶残基脱氨基形成尿嘧啶，导致DNA复制后胞嘧啶到胸腺嘧啶的转换突变。我们已在体外利用此反应破坏质粒pUC4K的氨基糖苷3'-磷酸转移酶I编码区中限制性内切酶HindIII和XmaI的识别序列。该方法应适用于产生含胞嘧啶单链末端的任何限制性内切酶识别序列的突变。文中还讨论了在编码区将限制性酶切位点诱变产生终止密码子的可能性。
