title: A frequency-controlled random mutagenesis method for GC-rich genes.
abstract: A novel random mutagenesis strategy was developed by combining sodium bisulfite modification with polymerase chain reaction (PCR). This method introduced the predominant substitution of GC to AT, meaning that it was more suitable for mutagenesis of GC-rich genes and helped to decrease the GC content of target DNA. Mutation efficiency correlated with modification time and different mutation frequency could easily be obtained by controlling modification time. The results indicated that this method could yield a desired and adequate frequency of random mutation to the DNA of interest, especially GC-rich genes, and provided a powerful tool for directed molecular evolution.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, People's Republic of China.
authors.forename: Pengfu
authors.initials: P
authors.lastname: Liu
authors.forename: Yuzhi
authors.initials: Y
authors.lastname: Hong
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Lin
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Fu
authors.forename: Ling
authors.initials: L
authors.lastname: Lin
authors.forename: Congcong
authors.initials: C
authors.lastname: Li
authors.forename: Xin
authors.initials: X
authors.lastname: Meng
authors.forename: Ziduo
authors.initials: Z
authors.lastname: Liu
mesh_terms: D020862:GC Rich Sequence / Q000235:genetics*; D016296:Mutagenesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013447:Sulfites / Q000737:chemistry
chemical_list: D013447:Sulfites; C042345:hydrogen sulfite