title: Splicing together different regions of a gene by modified polymerase chain reaction-based site-directed mutagenesis.
abstract: A modified polymerase chain reaction (PCR)-based site-directed mutagenesis method used to splice together different regions of a gene by deleting hundreds of nucleotides of undesired sequences is described. This method was inspired by a PCR-based site-directed mutagenesis method developed by Stratagene (La Jolla, CA, USA); the procedure and primer design were modified to enable the method to generate deletions several hundreds of nucleotides in length with an efficiency of 80-100%, and to delete two DNA fragments simultaneously in a single PCR. This method should be useful for deletion of large DNA fragments from a gene.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai, 200232, People's Republic of China.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Li
authors.forename: Yafeng
authors.initials: Y
authors.lastname: Qiu
authors.forename: Yang
authors.initials: Y
authors.lastname: Shen
authors.forename: Chan
authors.initials: C
authors.lastname: Ding
authors.forename: Peihong
authors.initials: P
authors.lastname: Liu
authors.forename: Jinping
authors.initials: J
authors.lastname: Zhou
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Ma
mesh_terms: D000818:Animals; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D017353:Gene Deletion; D006801:Humans; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA