基于快速定点突变技术的药学专业实验设计

Design of pharmacy experimental teaching project based on fast site-directed mutagenesis techniques

聚合酶链式反应(polymerase chain reaction,PCR)介导的快速定点突变技术是被广泛用于药物靶标发现及确证的关键技术,该技术可按照预定设计,对某些已知基因的指定碱基进行定点增删或替换。通过将PCR介导的快速定点突变这一新技术引入到药学专业药物化学生物学本科实验教学环节,设计了包含引物设计、PCR、核酸电泳及序列鉴定四个操作模块的定点碱基置换的教学综合实验。教学实践表明该实验拓展了药物化学生物学本科实验教学内容,加深了学生对靶标蛋白定点修饰和改造的认识与了解,培养了学生对复杂问题的主动探索精神,提高了学生理解运用先进技术解决实际问题的综合能力。

Fast site-directed mutagenesis is a new rapid and efficient polymerase chain reaction(PCR)-based site-directed mutagenesis method, which can do insertion, single-point mutation, and deletion rapidly and efficiently. After integrate this technique into the existing medicinal chemical biology experimental curriculum, a laboratory-based course with the goal of achieving DNA point substitution is designed. This laboratory-based class composed with four operation parts including the primer design, PCR, gel electrophoresis and sequence identification. Students’ feedback showed this student-centered and inquiry-based instruction laboratory class provide students with first-hand experience of drug targets modification and with the opportunity to explore methods used by scientists in their discipline, help students to gain new knowledge and understanding of group work with questions, improve students’ abilities in understanding and application of advanced technology for problem solving.