Semliki Forest virus capsid protein expressed by a baculovirus recombinant.
We have constructed a recombinant baculovirus which expressed the Semliki Forest (SFV) capsid (C) gene as a fusion protein under the control of the polyhedrin gene promoter. The sequence coding for C and part of the envelope E3 region were expressed as a polyprotein precursor. Spodoptera frugiperda (Sf9) insect cells infected with the recombinant virus produced a protein reacting with polyclonal rabbit antiserum that had been raised against the 33 kDa authentic C protein purified from SFV. Primer extension analyses showed that transcription from the polyhedrin promoter started in the late/very late consensus initiation motif. Due to its autoprotease activity, the recombinant C protein was contranslationally cleaved within its C-terminus and the mature protein remained undegraded for long periods of time. Spodoptera frugiperda cells infected with recombinant virus synthesized large amounts of C protein. The recombinant protein was karyophilic as is authentic SFV C protein and it was translocated into the nucleus; there, it was associated with nucleolus-like structures.
Favre
D
D
Institute of Medical Microbiology, University of Berne, Switzerland.
Studer
E
E
Nishimura
T
T
Weitz
M
M
Michel
M R
MR
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics*; D002451:Cell Compartmentation; D010450:Endopeptidases / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology / Q000382:microbiology; D000077183:Occlusion Body Matrix Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins; D014771:Virion / Q000235:genetics
D000077183:Occlusion Body Matrix Proteins; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D010450:Endopeptidases