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United States Department of Agriculture

Agricultural Research Service

Title: Expression of the Green Fluorescent Protein Carried by Autographa Californica Multiple Nucleopolyhedrosis in Insect Cell Lines

Authors
item Grasela, James
item McIntosh, Arthur
item Goodman, Cynthia
item Wilson, Louise - OXFORD BROOKES UNIV, UK
item King, Linda - OXFORD BROOKES UNIV, UK

Submitted to: National Entomological Society of America Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: August 6, 1999
Publication Date: N/A

Technical Abstract: A recombinant AcMNPV containing the green fluorescent protein gene (gfp) under the polyhedrin promoter was used to investigate the expression of the gfp gene as well as the production of recombinant extracellular virus in nine continuous insect cell lines including Heliothis virescens (BCIRL-HV- AM1), Helicoverpa zea (BCIRL-HZ-AM1), Anticarsia gemmatalis (BCIRL-AG-AM1), ,Trichoplusia ni (TN-CL1), Spodoptera frugiperda (IPLB-SF21), its clone Sf9 Spodoptera exigua (Se-E1; Se-E5), Bombyx mori (BMN), and five cell line clones of BCIRL-HV-AM1. Of the 14 cell lines tested, all were permissive with varying degrees to AcMNPV.GFP except BCIRL-HV-AMCL2, BCIRL-HZ-AM1, and BMN which were nonpermissive to the virus. Except for Se-E1, IPLB-SF21 and 4 of the 5 BCIRL-HV-AM1 clones, all the other cell lines began to express GFP at 48 h postinoculation in ExCell401 media. Se-E1 in serum-containing medium expressed the GFP at 48 h postinoculation. TN-CL1 cells produced the largest mean percent number of fluorescent cells both in serum-containing (76.6%) and serum-free medium (64.8%) at 120 h postinoculation. All the BCIRL-HV-AM1 clones showed no GFP expression until 96 h postinoculation, and only then about 1% of the cell population fluoresced. ECV production at 120 h postinoculation was highest in Se-E5 cells grown in serum-containing medium (37.8 TCID50/ml x 10**6) followed by BCIRL-HV-AM1 in TC199-MK (33.4 TCID50/ml x 10**6). Only BCIRL-HV-AMCL3 produced any substantial level of ECV at 120 h postinoculation (16.9 TCID50/ml x 10**6). There was no significant correlation between ECV production and the mean percent number of fluorescent cells. This study provides further information on the susceptibility of 14 insect cell lines to a recombinant AcMNPV containing the green fluorescent protein gene.

Last Modified: 7/24/2014
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