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.
Russo
S
S
Institute of Microbiology and Immunology, Faculty of Veterinary Medicine, Milan, Italy.
Montermini
L
L
Berkovitz-Siman-Tov
R
R
Ponti
W
W
Poli
G
G
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