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United States Department of Agriculture

Agricultural Research Service

Title: Assessment of the Permissive Nature of Insect Cell Lines from Various Orders Challenged with Acmnpv Carrying a Fluorescent Marker Gene

Authors
item McIntosh, Arthur
item Grasela, James
item Popham, Holly

Submitted to: Baculovirus and Insect Cell Culture Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: January 12, 2005
Publication Date: February 21, 2005
Citation: Mcintosh, A.H., Grasela, J.J., Popham, H.J. 2005. Assessment of the permissive nature of insect cell lines from various orders challenged with acmnpv carrying a fluorescent marker gene [abstract]. Baculovirus and Insect Cell Culture Conference Proceedings. p. 31.

Technical Abstract: Insect cell lines from Arthropoda represented by Lepidoptera, Coleoptera, Diptera and Homoptera were evaluated for their ability to support replication of AcMNPV. In addition, some of the cell lines that were refractive to AcMNPV were tested with AcMNPVhsp70 Red, a recombinant carrying the red fluorescent protein (RFP), for their ability to express this protein following inoculation. Of the 9 lepidopteran cell lines tested, only 3 cell lines from Helicoverpa zea (BCIRL -HZ-AM1), Lymantri dispar ( IPLB-LD65) and Cydia pomonella (CP-169) failed to support appreciative viral replication as measured by TCID50 assay. Heliothis virescens (BCIRL-HV-AM1) produced the highest viral titer of 2.32 x 10**7 + or - 5.19 x 10**-2 followed by Heliothis subflexa (BCIRL-HS-AM1) of 4.69 x 10**6 + or - 6.48 x 10**-2 and Spodoptera frugiperda (IPLB-SF21) with 4.13 x 10**6 + or - 10**-2. None of the Coleopteran, Dipteran or Homopteran cell lines supported AcMNPV replication. However, when studies were performed employing AcMNPVhsp70 Red, the Dipteran cell lines Aedes aegypti (ATC-10) and Drosophila melanogaster (Line2) both expressed the RFP as well as the refractive Lepidopteran cell lines from H. zea and L. dispar. No RFP expression was observed in any of the Coleopteran or Homopteran cell lines. Cell lines refractive to AcMNPV did not appear to be adversely affected by the virus as judged by their ability to multiply, nor was there any indication of induced apoptosis as assessed by DNA fragmentation profiles.

Last Modified: 11/28/2014
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