title: Multiple Site-Directed and Saturation Mutagenesis by the Patch Cloning Method.
abstract: Constructing protein-coding genes with desired mutations is a basic step for protein engineering. Herein, we describe a multiple site-directed and saturation mutagenesis method, termed MUPAC. This method has been used to introduce multiple site-directed mutations in the green fluorescent protein gene and in the moloney murine leukemia virus reverse transcriptase gene. Moreover, this method was also successfully used to introduce randomized codons at five desired positions in the green fluorescent protein gene, and for simple DNA assembly for cloning.
authors.affiliation: PeptiDream Inc., 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8505, Japan.
authors.forename: Naohiro
authors.initials: N
authors.lastname: Taniguchi
authors.affiliation: Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan. murah@apchem.nagoya-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Murakami
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D003062:Codon / Q000235:genetics; D004247:DNA / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D008979:Moloney murine leukemia virus / Q000235:genetics; D016296:Mutagenesis / Q000235:genetics*; D016297:Mutagenesis, Site-Directed / Q000379:methods; D009154:Mutation / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods; D012194:RNA-Directed DNA Polymerase / Q000235:genetics
chemical_list: D003062:Codon; D049452:Green Fluorescent Proteins; D004247:DNA; D012194:RNA-Directed DNA Polymerase
keywords: Cloning; Protein engineering; Saturation mutagenesis; Site-directed mutagenesis