title: Internal epitope tagging informed by relative lack of sequence conservation.
abstract: Many experimental techniques rely on specific recognition and stringent binding of proteins by antibodies. This can readily be achieved by introducing an epitope tag. We employed an approach that uses a relative lack of evolutionary conservation to inform epitope tag site selection, followed by integration of the tag-coding sequence into the endogenous locus in zebrafish. We demonstrate that an internal epitope tag is accessible for antibody binding, and that tagged proteins retain wild type function.
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, United States.
authors.forename: Leonard
authors.initials: L
authors.lastname: Burg
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, United States.
authors.forename: Karen
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, United States.
authors.forename: Tristan
authors.initials: T
authors.lastname: Bonawitz
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, United States.
authors.forename: Viktorija
authors.initials: V
authors.lastname: Grajevskaja
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, United States.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Bellipanni
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, United States.
authors.forename: Richard
authors.initials: R
authors.lastname: Waring
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, United States.
authors.forename: Darius
authors.initials: D
authors.lastname: Balciunas
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics*; D000939:Epitopes / Q000235:genetics*; D011506:Proteins / Q000235:genetics; D016415:Sequence Alignment / Q000379:methods; D015027:Zebrafish
chemical_list: D000906:Antibodies; D000939:Epitopes; D011506:Proteins