Larionov O A, Nikiforov V G
Genetika. 1982 Mar;18(3):349-59.
Methods for generating mutations in preselected genes of prokaryotes and eukaryotes are reviewed. Directed mutagenesis is based on mutagenic treatment of genome fragments in vitro instead of a whole genome in vivo. The cloning methods make it possible to perform directed mutagenesis even using conventional nonspecific mutagens. Methods have been elaborated for selective modification of specific sequences in cloned genes, based on the specific DNA recognition properties of restriction endonucleases and complementary poly- and oligonucleotides. Chemically synthesized oligonucleotides may be used for constructing mutant genes containing any base substitutions, small insertions and deletions.
本文综述了在原核生物和真核生物的预选基因中产生突变的方法。定向诱变是基于对体外基因组片段而非体内全基因组进行诱变处理。克隆方法使得即使使用常规非特异性诱变剂也能够进行定向诱变。基于限制性内切酶以及互补的多核苷酸和寡核苷酸的特定DNA识别特性,已经开发出了对克隆基因中的特定序列进行选择性修饰的方法。化学合成的寡核苷酸可用于构建包含任何碱基替换、小片段插入和缺失的突变基因。