Expression of bovine leukemia virus ENV glycoprotein in insect cells by recombinant baculovirus

FEBS Letters

Berkovitz-Siman-Tov, Revital²  Ponti, Wilma³  Montermini, Laura¹  Poli, Giorgio³  Russo, Silvia³ 

[1]Department of Neurology, Centre de Recherche L.C. Simard, Hôpital Notre Dame, Institute du Cancer, 1560 rue Sherbrooke East, Montreal, Que. H2L 4MI, Canada

[2]Department of Post-Harvest Science of Fresh Products, ARO, The Volcani Center, Bet Dagan 50250, Israel

[3]Institute of Microbiology and Immunology, Faculty of Veterinary Medicine, Via Celoria, 10, 20133 Milan, Italy

关键词: Baculovirus;    Biotechnology;    Bovine leukemia virus;    Glycosylation;    Recombinant glycoprotein;    AcMNPV;    Autographa californica multiple nuclear polyhedrosis virus;    BCA;    bichinchoninic acid protein quantitative assay;    BEVS;    baculovirus expression vector system;    BLV;    bovine leukemia virus;    bp;    base pair;    BSA;    bovine serum albumin;    DEPC;    diethyl-pyrocarbonate;    FCS;    fetal calf serum;    FLK;    fetal lamb kidney cell line;    FLK-gp51;    gp51 purified from persistently infected FLK cell;    gp51-p30;    glycoproteins constituting BLV envelope;    kbp;    103 base pairs;    kD;    103 daltons;    MAb;    monoclonal antibody;    MOI;    multiplicity of infection;    occ −;    AcMNPV plaques showing polh−phenotype;    PBS;    phosphate buffered saline;    PCR;    polymerase chain reaction;    p.i.;    post infection;    polh;    polyhedrin coding gene of AcMNPV;    PRINS;    primed in situ labelling of nucleic acids;    SDS-PAGE;    sodium dodecylsulfate polyacrylamide gel electrophoresis;   

DOI  :  10.1016/S0014-5793(98)00951-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The gp51-p30 glycoprotein constituting BLV envelope was expressed in Sf-21 insect cells by means of recombinant baculoviruses. Post-infection cell lysates were analyzed, in order to define the immunologic reactivity of recombinant products. Oligosaccharide chains, containing N-acetylglucosamine, mannose, galactose and sialic acid were found on recombinant gp51-p30. In order to investigate the timing of transcription and translation of the glycoprotein, kinetic assays were carried out on cell lysates and directly in situ on Sf-21 cells during the course of baculovirus infection. The use of different solubilizing reagents was also evaluated in order to rescue recombinant glycoprotein from its subcellular location.

【 授权许可】

Unknown