title: A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC.
abstract: Not all proteins are tolerable to mutations. Whether a specific protein can be a mutable target is of importance in the biotechnology and pharmaceutical industry. This study reported a novel mutagenesis assay using tandem NNT and NNC oligonucleotides to test the mutability of a candidate gene. These two tandem oligonucleotides avoid the risk of forming nonsense mutations and render flexibility of truncating or expanding the insertion size. As a reporter gene, ZeoR (zeocin resistance gene) was confirmed to have a high tolerance for mutagenesis by this new assay.
authors.affiliation: Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
authors.forename: Huifen
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Human Anatomy, University of South China, Hengyang 421001, China.
authors.forename: Cuilan
authors.initials: C
authors.lastname: Zhou
authors.affiliation: Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
authors.forename: Andy K
authors.initials: AK
authors.lastname: Zhang
authors.affiliation: Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D017930:Genes, Reporter / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics*; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids / Q000235:genetics
chemical_list: D009841:Oligonucleotides