Selection of Peptide Ligands Specific for Baculovirus DNA-Binding Protein from the Flitrx~(TM) Random Peptide Display Library

摘要

Autographa californica nucleopolyhedrovirus (AcNPV) is a baculovirus that is widely employed as a vector for the expression of foreign genes and pest control.Although baculoviruses,including AcNPV,efficiently replicate in the nuclei of arthropod cells,the dynamics and mechanism of DNA replication within the infected cell are still poorly understood.It has been found that the DNA-binding protein (DBP) is an early gene product and appears to be crucial for viral DNA replication.Presented here is the selection of peptide ligands that specifically bind to DBP for AcNPV from the FliTrx~(TM) random peptide display library;this entails the amplification,cloning of the DNA-binding protein (DBP) gene from AcNPV and the construction of the expression plasmid for DBP,and the expression and purification of the recombinant His.Tag AcNPV DBP that was used as a target molecule for the selection of the peptide ligands specific for AcNPV DBP.The affinity and efficiency of such peptide ligands were then measured by ELISA procedures.The beneficial aspect of this research is the monospecificity quality of the peptide ligands specific for AcNPV DBP.They could be used for the study of the dynamics of the viral genome and its replication within the infected cell,for the development of a quantitative method for the determination of the presence of baculovirus in various samples,for the development of a peptide ligand-based assay for the determination of baculovirus titers;or they could be immobilized on a chromatographic support for an improved affinity purification of AcNPV DBP.