Recombinant protein production in insect larvae: host choice, tissue distribution, and heterologous gene instability.
The expression of the fluorescent protein, DsRed, facilitates the optimization of protein production in orally-infected whole larvae. Trichoplusia ni was shown to be a much better host for recombinant AcMNPV compared to four other noctuid Lepidopteran species achieving 100% infectivity at the minimal tested dose. The highest density of marker protein was found in endothelial and tracheal cells, fat body, and hemocytes. Trichoplusia ni larvae possessed visually detected color over sequential passages of oral infection until the sixth round. Western blot analysis confirmed the progressive decrease of both tetramer and monomer forms of DsRed. The intact DsRed gene and promoter region was present in late passages, but viral population carrying the heterologous gene had dropped more than 2-logs after the fifth round while the amount of total viral DNA remained unchanged over sequential passages.
Kovaleva
Elena S
ES
Chesapeake PERL Inc., Protein Expression and Recovery Laboratories, 8510A Corridor Road, Savage, MD 20763, USA. ekovaleva@c-perl.com
O'Connell
Kevin P
KP
Buckley
Patricia
P
Liu
Zhi
Z
Davis
David C
DC
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000254:growth & development*; D042783:Endothelial Cells / Q000737:chemistry; D005216:Fat Body / Q000737:chemistry; D006434:Hemocytes / Q000737:chemistry; D007814:Larva / Q000821:virology; D007915:Lepidoptera / Q000821:virology*; D008164:Luminescent Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014132:Trachea / Q000737:chemistry
D008164:Luminescent Proteins; D011994:Recombinant Proteins; C413663:fluorescent protein 583