摘要
对大引物PCR定点突变方法进行改进,主要包括:避免在一个PCR反应中同时使用扩增全长基因的常规引物,选择不同载体克隆原始基因和突变基因,通过抗性选择筛选突变子,从而完全避免原始基因的干扰.实验结果证明利用改进后的方法进行基因点突变,可以减少操作步骤,提高突变频率.
＂Megaprimer＂ PCR used to introduce site-mutations into a target gene was improved here, including mainly omitting using of a normal primer pair in one reaction for amplification of a full length gene, and choosing different vectors for cloning of wild type and mutant genes, respectively. Therefore, mutants were yielded efficiently by subsequent selection on antibiotics-containing medium, and sequencing analysis showed that interference of wild type genes was completely eliminated by this improved method.
出处
《湖北大学学报（自然科学版）》
CAS
北大核心
2009年第1期79-81,共3页
Journal of Hubei University：Natural Science
基金
国家自然科学基金(30770036)
教育部高等学校博士学科点专项科研基金(20070511004)
教育部留学回国人员科研启动基金资助
