Functional domain mapping using M13 deletions.
We describe a convenient approach for concomitant functional characterization of peptide domains (monoclonal antibody epitopes, receptor-ligand, DNA-protein, and protein-protein interaction sites, etc) encoded by the sequential series of overlapping M13 subclones generated for nucleotide sequence determination of a new cDNA. We have employed this method to rapidly map the location and amino acid sequence of an epitope-containing domain within a polypeptide encoded by a newly isolated cDNA.
Department of Anatomy and Cellular Biology, Tufts University School of Medicine, Boston, MA 02111, USA.
N
N
Grammatikakis
B P
BP
Toole
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry; D017104:Bacteriophage M13 / Q000201:enzymology / Q000235:genetics; D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary; D004587:Electrophoresis, Agar Gel; D000939:Epitopes / Q000032:analysis* / Q000737:chemistry; D005814:Genes, Viral; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D017384:Sequence Deletion; D001616:beta-Galactosidase / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D000939:Epitopes; D010455:Peptides; D011506:Proteins; D011994:Recombinant Proteins; D001616:beta-Galactosidase