Secretion of green fluorescent protein from recombinant baculovirus-infected insect cells.
Trichoplusia ni (High Five) and Spodoptera frugiperda (Sf21) cells were engineered for expression of epitope (Flag)-tagged signal peptide-green fluorescent protein (GFP) fusions to examine the suitability of GFP as a secretory marker. The recombinant baculovirus-infected cells became fluorescent, and the High Five cells but not Sf21 cells secreted GFP in the culture medium as detected by the presence in the culture supernatant of a Flag-immunoreactive 30-kDa species and the characteristic 510-nm GFP fluorescence peak. Signal peptides derived from ecdysteroid UDP-glucosyltransferase of Autographa californica nuclear polyhedrosis virus and from rat brain glutamate receptor were both able to promote secretion of GFP. GFP may thus be used as a research tool in the study of the secretory process in insect cells both in cell biology and in biotechnological applications.
Laukkanen
M L
ML
VTT Biotechnology and Food Research, Espoo, Finland.
Oker-Blom
C
C
Keinänen
K
K
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D015870:Gene Expression; D005964:Glucosyltransferases / Q000096:biosynthesis; D049452:Green Fluorescent Proteins; D007313:Insecta; D008164:Luminescent Proteins / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000096:biosynthesis; D051381:Rats; D017470:Receptors, Glutamate / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D016324:Sequence Tagged Sites; D018411:Spodoptera; D014162:Transfection
D017931:DNA Primers; D008164:Luminescent Proteins; D021382:Protein Sorting Signals; D017470:Receptors, Glutamate; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005964:Glucosyltransferases; C086659:ecdysteroid UDP-glucosyltransferase