Ex vivo monitoring of protein production in baculovirus-infected Trichoplusia ni larvae with a GFP-specific optical probe.
Trichoplusia ni larvae were infected with baculoviruses containing genes for the expression of ultraviolet optimized green fluorescent protein (GFPuv) and several product proteins. A GFP-specific optical probe was used to both excite the green fluorescent protein (lambda(ex) = 385 nm), and subsequently monitor fluorescence emission (lambda(em) = 514 nm) from outside the infected larvae. The probe's photodetector was connected to a voltmeter, which was used to quantify the amount of GFPuv expressed in infected larvae. Voltage readings were significantly higher for infected vs. uninfected larvae and, by Western analysis, linear with the amount of GFPuv produced. In addition, the probe sensitivity and range were sufficient to delineate infection efficiency and recombinant protein production for model proteins, chloramphenicol acetyltransferase and human interleukin-2. This work represents a critical step in developing an automated process for the production of recombinant proteins in insect larvae.
Kramer
Shannon F
SF
Center for Biosystems Research, University of Maryland Biotechnology Institute, and Department of Chemical Engineering, University of Maryland, College Park, Maryland 20742, USA.
Kostov
Yordan
Y
Rao
Govind
G
Bentley
William E
WE
D000818:Animals; D016367:Baculoviridae / Q000472:pathogenicity*; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D007814:Larva / Q000821:virology; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D009036:Moths / Q000821:virology*; D013050:Spectrometry, Fluorescence / Q000379:methods; D018411:Spodoptera / Q000254:growth & development; D014162:Transfection / Q000379:methods
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D002374:Catalase