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Characterization of the Epitope Recognized by a Monoclonal Antibody Directed against the Largest Subunit ofDrosophilaRNA Polymerase II by Roland E. Kontermann; Zhihong Liu; Rüdiger A. Schulze; Kai A. Sommer; Iris Queitsch; Stefan Dübel; Sergey M. Kipriyanov; Frank Breitling; Ekkehard K.F. Bautz is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Characterization of the Epitope Recognized by a Monoclonal Antibody Directed against the Largest Subunit ofDrosophilaRNA Polymerase II about?
The epitope recognized by monoclonal antibody MAb215 generated previously against Drosophila melanogaster RNA polymerase II was mapped to amino acid residues 806-820 of the largest, 215 kDa, subunit located in a region conserved within the largest subunits of pro-and eukaryotic RNA polymerases. The affinities of MAb215 and of a recombinant single-chain Fv fragment (scFv215) were determined for binding to the enzyme as well as the fusion protein and synthetic peptides used for epitope mapping. In addition, amino acid residues of the epitope important for binding to MAb215 were identified using peptides carrying single amino acid substitutions. The epitope is not involved in the polymerization reaction or the DNA unwinding process since no inhibitory effects of the monoclonal antibody were observed in nonspecific in vitro transcription using denatured calf thymus DNA or double stranded oligo dC-tailed T7 DNA as template. In contrast, MAb215 inhibits accurate in vitro transcription from the Krüppel gene promoter and from the adenovirus-2 major late promoter. Preincubation of template DNA with the nuclear extract had no effects on inhibition supporting the notion that the epitope does
Who reads Characterization of the Epitope Recognized by a Monoclonal Antibody Directed against the Largest Subunit ofDrosophilaRNA Polymerase II?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Roland E. Kontermann; Zhihong Liu; Rüdiger A. Schulze; Kai A. Sommer; Iris Queitsch; Stefan Dübel; Sergey M. Kipriyanov; Frank Breitling; Ekkehard K.F. Bautz
- Published
- 1995
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)