Expression of recombinant E2 and C proteins of rubella virus in insect cells.
We have constructed a recombinant baculovirus expressing the rubella virus E2 (42-45 KDa) and C (34 KDa) proteins. Sf9 cells infected with recombinant virus were able to synthesize and process the two proteins coded by a unique precursor gene. By immunoblot and immunoprecipitation analysis with polyclonal and monoclonal antibodies, a precursor polyprotein (66 KDa) and two other proteins migrating with an apparent molecular weight of 42 KDa and 36 KDa were recognized as E2 glycoprotein and C protein, respectively. The recombinant E2 protein appeared to be glycosylated since it was susceptible to tunicamycin. The results indicate that the RV polyprotein coding for E2 and C is expressed and proteolytically cleaved in insect cells. This baculovirus expression system provides a useful alternative approach for the production of rubella virus antigens and should allow the purification of large quantities of the RV proteins for further biochemical and immunological studies.
Cusi
M G
MG
Department of Molecular Biology, University of Siena, Italy.
Bianchi
S
S
Cioè
L
L
Valensin
P E
PE
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006031:Glycosylation; D015151:Immunoblotting; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D012412:Rubella virus / Q000378:metabolism*; D018411:Spodoptera; D014162:Transfection; D014758:Viral Core Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification
D000906:Antibodies; D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; C074573:nucleocapsid protein (C), rubella virus; C061827:E2 envelope protein, Rubella virus