Expression of two heterologous proteins depends on the mode of expression: comparison of in vivo and in vitro methods.
The yield of two proteins, avidin and green fluorescent protein (GFP), expressed from a modified Autographa californica nucleopolyhedrovirus (AcMNPV), was compared in Sf9 cell culture monolayer, Sf21 cell suspension culture and intact Spodoptera litura larvae. GFP expressed from the p10 promoter yielded up to 1.5% of total soluble protein in larvae, 20-fold higher than that in monolayer suspension culture. Avidin, expressed from the polh promoter, yielded up to 2.3% of total soluble protein in larvae, 10-fold higher than that in suspension culture and 40-fold higher than that in monolayers. Avidin expression did not affect amounts of GFP in dual-expressing baculovirus compared with those detected from a GFP-only expressing AcMNPV. A biotin-binding assay showed that all avidin expressed in larvae was fully active. Glycosylation patterns of chicken-avidin and Spodoptera-avidin were very similar, though the latter showed a proportion of partially glycosylated material.
Gatehouse
Laurence N
LN
Horticulture and Food Research Institute of NZ, Tennant Drive, Palmerston North 4474, New Zealand. lgatehouse@hortresearch.co.nz
Markwick
Ngaire P
NP
Poulton
Joanne
J
Young
Vivienne L
VL
Ward
Vernon K
VK
Christeller
John T
JT
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D005786:Gene Expression Regulation / Q000502:physiology; D005822:Genetic Vectors / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism*; D018411:Spodoptera / Q000502:physiology*
D011994:Recombinant Proteins