Varga-Orvos Zoltán, Nagy Zsolt B, Mészáros Attila, Kökény Szabolcs, Gergely Péter, Tamás László, Poór Gyula
Laboratory of Functional Genomics, Biological Research Centre, Hungarian Academy of Sciences, P.O. Box 521, Szeged, 6701, Hungary.
Biotechnol Lett. 2007 Dec;29(12):1921-5. doi: 10.1007/s10529-007-9474-7. Epub 2007 Aug 9.
Site-directed mutagenesis is of great importance for probing the structure/function relationship of proteins. Developing our previous method (Nagy et al. Anal Biochem 324:301-303, 2004), here we report a multiplex strategy for site-directed mutagenesis using PCR in one tube to introduce a single mutation into three or more genes at the same time. DNA fragments carrying the desired mutation can be distinguished from each other in a standard antibiotic selection step of the transformed bacteria. Due to this strategy the mutagenesis procedure for several genes can be accelerated.
定点诱变对于探究蛋白质的结构/功能关系非常重要。在我们之前方法（Nagy等人，《分析生物化学》324:301 - 303，2004年）的基础上，我们在此报告一种在一管中使用聚合酶链式反应（PCR）进行定点诱变的多重策略，可同时将单个突变引入三个或更多基因。在转化细菌的标准抗生素筛选步骤中，可以区分携带所需突变的DNA片段。由于该策略，几个基因的诱变过程可以加快。
