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.
Department of Molecular Biology, University of Siena, Italy.
M G
MG
Cusi
S
S
Bianchi
L
L
Cioè
P E
PE
Valensin
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