Characterisation of an avian influenza virus nucleoprotein expressed in E. coli and in insect cells.
The nucleoprotein (NP) gene from influenza virus A/Shearwater/Australia/72 has been expressed intracellularly in both E. coli and insect cells. E. coli-derived NP was identified by Western blot analysis as a 56 kDa protein which co-migrates with virion-derived NP. This protein was purified by immunoaffinity chromatography and a nitrocellulose binding assay showed that NP formed complexes with positive- and negative-sense influenza neuraminidase RNA transcribed in vitro. ELISA and Western blot analysis revealed that recombinant NP of 56 kDa was produced in high yields in insect cells using a baculovirus vector. Immunofluorescence microscopy revealed that NP was localised to the nucleus of infected insect cells.
Harley
V R
VR
CSIRO Division of Biomolecular Engineering, Parkville, Victoria, Australia.
Mather
K A
KA
Power
B E
BE
McKimm-Breschkin
J L
JL
Hudson
P J
PJ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D009980:Influenza A virus / Q000235:genetics / Q000378:metabolism*; D007313:Insecta / Q000166:cytology; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000235:genetics; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D016372:RNA, Antisense / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014758:Viral Core Proteins; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism
C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D016372:RNA, Antisense; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D014764:Viral Proteins; D009439:Neuraminidase