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Title: BACULOVIRUS-BASED EXPRESSION OF AN INSECT VIRAL PROTEIN IN 12 DIFFERENT INSECT CELL LINES.

Author
item Chen, Yanping - Judy
item Gundersen-Rindal, Dawn
item Lynn, Dwight

Submitted to: In Vitro Cellular and Developmental Biology - Animal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/11/2005
Publication Date: 2/11/2005
Citation: Chen, Y.P., Gundersen-Rindal, D.E., Lynn, D.E. 2005.Baculovirus-based expression of an insect viral protein in 12 different insect cell lines. In Vitro Cellular and Developmental Biology - Animals.41(1):43-49.

Interpretive Summary: Viruses that attack insects are becoming an important tool in insect control programs. Evaluating the ability of viruses to kill insects, though, can be difficult if live insects are required for testing. Cell lines, derived from insect tissues, have proven to be a useful alternative in virus testing, eliminating the need for live insects. We determined the ability of 12 different insect cell lines to support virus growth and found all to be suitable. The results of this research demonstrate that a wide variety of cell lines can be used to grow viruses. This information will be used by other scientists involved in insect virus research and by companies that market viruses for insect control.

Technical Abstract: The ability of 12 unique lepidopteran insect cell lines from Anticarsia gemmatalis, Heliothis virescens, Lymantria dispar (two lines), Mamestra brassica, Plutella xylostella, Spodoptera frugiperda (two lines), and Trichoplusia ni (three lines) to support production of a recombinant polydnavirus protein (GiPDV 1.1) expressed using the Bac-to-Bac' baculovirus expression system was examined. Polydnavirus gene GiPDV 1.1 was cloned into the pFastBac baculovirus vector under the control of the polyhedron promoter, followed by generation of recombinant bacmid-GiPDV 1.1 by site-specific transposition. The ability of each insect cell line to support recombinant polydnavirus gene expression was estimated using RT-PCR and Western blot. Each insect cell line infected with recombinant bacmid-GiPDV 1.1 and tested in this study was capable of supporting and producing recombinant protein. Time course expression analysis showed 72 to 96 hour post transfection to be the optimal time for harvest of recombinant protein for each insect cell line.