Expression of bovine leukemia virus ENV glycoprotein in insect cells by recombinant baculovirus.
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.
Institute of Microbiology and Immunology, Faculty of Veterinary Medicine, Milan, Italy.
S
S
Russo
L
L
Montermini
R
R
Berkovitz-Siman-Tov
W
W
Ponti
G
G
Poli
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D006031:Glycosylation; D002479:Inclusion Bodies / Q000737:chemistry; D007313:Insecta / Q000821:virology*; D007700:Kinetics; D037102:Lectins / Q000378:metabolism; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D012995:Solubility; D013379:Substrate Specificity; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
D037102:Lectins; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; D014759:Viral Envelope Proteins; C060711:gp30 envelope transmembrane protein, Bovine leukemia virus