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Inorganic binding peptide‐mediated immobilization based on baculovirus surface display system
Author(s) -
Song Lei,
Liu Yingying,
Chen Jinchun
Publication year - 2010
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.200900359
Subject(s) - autographa californica , sf9 , recombinant dna , nuclear polyhedrosis virus , gene delivery , microbiology and biotechnology , peptide , gene , mcherry , biophysics , biology , infectivity , chemistry , transfection , virus , virology , green fluorescent protein , biochemistry , spodoptera
The biomolecule‐mediated assembly of novel composites has been the subject of numerous investigations during the last years, providing new insights into material science and engineering. Via molecular biology technology, we were able to introduce the genetically engineered polypeptide for inorganics (GEPI) as a molecular binder into biomolecules such as phage viruses, to assemble hybrid functional nanoarchitectures. In the present work, we introduced a novel nanocomposite comprising the Autographa californica nuclear polyhedrosis virus (AcNPV) and nanoparticles bound to it. Our results show that a GEPI‐encoding gene was successfully introduced by recombination into a eukaryotic expression bacmid and finally displayed outside of the AcNPV after transfection into Sf9 insect cells using the Bac‐to‐Bac baculovirus expression system. The recombinant baculovirus maintained both the viral infectivity and the specific binding activity of the GEPI. The construction of the gene in the recombinant plasmid was examined by polymerase chain reaction analysis and enzymatic digestion identification, and verified by gene sequencing. Surface display of the fused peptide was revealed by Western blot analysis in dissolution studies and determined by immuno‐ gold electron microscopy. Adherence of nanoparticles to the recombinant baculovirus was visualized by transmission electron microscopy analysis. Here, we demonstrated the possibilities of combining peptide‐mediated immobilization with baculovirus surface display technology. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)