Expression of influenza neuraminidase in baculovirus-infected cells.
Recombinant influenza neuraminidase (NA, subtype 2, A/NT/60/68) was produced by recombinant baculovirus-infected insect cells. The recombinant NA retained enzyme activity and was located on the cell surface. Enzyme activity was both cell-associated and in the cell free supernatant; maximal NA activity was found in the supernatant. Recombinant NA was recognised by polyclonal antisera and by three monoclonal antibodies specific for NA (subtype 2). Enzyme activity was also neutralised by polyclonal antisera. Recombinant NA thus retains most of the immunological and activity properties of authentic influenza NA. Immunoprecipitation of [35S]Methionine-labelled cells and supernatant and partial purification of NA indicated that a approximately 50-kDa form of NA was present in the supernatant, whilst the expected size (approximately 67-kDa) was cell-associated. Purified recombinant extracellular virus was also enzymatically active, and contained the 67-kDa NA which was located on the membrane capsule of the virus. This suggests that the virus had acquired the cell-associated form of recombinant NA during the budding process from the cell.
Mather
K A
KA
Biomolecular Research Institute, Parkville, Australia.
White
J F
JF
Hudson
P J
PJ
McKimm-Breschkin
J L
JL
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D015870:Gene Expression; D007313:Insecta; D008854:Microscopy, Electron; D009439:Neuraminidase / Q000235:genetics* / Q000302:isolation & purification; D009975:Orthomyxoviridae / Q000201:enzymology* / Q000235:genetics* / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D011995:Recombination, Genetic
D011994:Recombinant Proteins; D009439:Neuraminidase