tailieunhanh - Báo cáo khoa học: Altering the surface properties of baculovirus Autographa californica NPV by insertional mutagenesis of the envelope protein gp64

The envelope protein gp64 of the baculovirusAutographa californicanuclear polyhedrosis virus is essential for viral entry into insect cells, as the glycoprotein bothmediates pH-dependentmembrane fusionandbinds tohost cell receptors. Surface modification of baculovirus particles by genetic engineering of gp64 has been demonstrated by various strategies and thus has become an important and powerful tool in molecular biology. | Eur. J. Biochem. 269 4458-4467 2002 FEBS 2002 doi Altering the surface properties of baculovirus Autographa californica NPV by insertional mutagenesis of the envelope protein gp64 Alexandra Spenger Reingard Grabherr Lars Tollner Hermann Katinger and Wolfgang Ernst Institute of Applied Microbiology University of Agricultural Sciences Vienna Austria The envelope protein gp64 of the baculovirus Autographa californica nuclear polyhedrosis virus is essential for viral entry into insect cells as the glycoprotein both mediates pH-dependent membrane fusion and binds to host cell receptors. Surface modification of baculovirus particles by genetic engineering of gp64 has been demonstrated by various strategies and thus has become an important and powerful tool in molecular biology. To improve further the presentation of peptides on the surface of baculovirus particles several insertion sites within the gp64 envelope protein were selected by their theoretical maximum surface probability and investigated for efficient peptide presentation. The ELDKWA peptide of the gp41 of HIV-1 specific for the human mAb 2F5 was inserted into 17 different positions of the glycoprotein gp64. Propagation of viruses was successful in 13 cases mutagenesis at four positions did not result in production of intact virus particles. Western blotting FACS analysis and ELISA were used for characterization of the different binding properties of the mutants. Insertion of this peptide into the native envelope protein resulted in high avidity display on the surface of baculovirus particles. This approach offers the possibility of effective modification of surface properties in regard to host range specificity and antigen display. Keywords Autographa californica nuclear polyhedrosis virus baculovirus ELDKWA epitope gp64 envelope protein surface display. The baculovirus Autographa californica nuclear polyhedrosis virus has been widely used as an expression system for .