Identification of epitope on DNA-binding protein expressed in insect cell infected by baculovirus.
DNA-binding protein (DBP) is an early gene product produced during viral replication. Polyclonal anti-DBP was produced using rabbit by intradermal injections of Escherichia coli-expressed purified recombinant DBP. Prepared anti-DBP completely blocked the replication of baculovirus in insect cells. The anti-DBP binding to DBP was confirmed by both Western blotting with Tn-5B1-4 insect cell lysates as well as immunostained baculovirus-infected Tn-5B1-4 insect cells. To determine the anti-DBP epitope 12 peptides were synthesized and their specific-binding activities were measured using ELISA. Based on specific-binding activity against anti-DBP the epitope was predicted to be between amino acid residues 248-265 (QRMSVEDFDRLFEMDKID). Especially from 18 amino acid residues it was further to be narrowed between amino acid residues 260-265 (EMDKID) which showed a critical role in specific-binding activity.
Suzuki
Mami
M
Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka, 422-8529, Japan.
Kohsaka
Tetsuya
T
Park
Enoch Y
EY
D000818:Animals; D000906:Antibodies / Q000502:physiology; D016367:Baculoviridae / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D000939:Epitopes / Q000737:chemistry*; D005822:Genetic Vectors / Q000235:genetics*; D007313:Insecta / Q000737:chemistry*; D008969:Molecular Sequence Data; D014779:Virus Replication
D000906:Antibodies; D004268:DNA-Binding Proteins; D000939:Epitopes