title: Rapid gene splicing and multi-sited mutagenesis by one-step overlap extension polymerase chain reaction.
abstract: Gene splicing and site-directed mutagenesis (SDM) are important to introduce desired sequences in target DNA. However, introducing mutations at multiple sites requires multiple steps of DNA manipulation, which is time-consuming and labor-intensive. Here, we present a rapid efficient gene splicing and multi-sited mutagenesis method that introduces mutations at two distant sites via sequential connection of DNA fragments by one-step overlap extension polymerase chain reaction (OE-PCR). This bottom-up approach for DNA engineering can be broadly used to study protein structure-function, to optimize codon use for protein expression, and to assemble genes of interest.
authors: Lauren Biotechnology, Nanjing 210019, China.
authors: Huilin
authors: H
authors: Wei
authors: Jie
authors: J
authors: Hu
authors: Lin
authors: L
authors: Wang
authors: Feng
authors: F
authors: Xu
authors: ShuQi
authors: S
authors: Wang
mesh_terms: D004274:DNA, Recombinant / Q000235:genetics*; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
chemical_list: D004274:DNA, Recombinant