Transduction of cultured fish cells with recombinant baculoviruses

• Published in: Journal of general virology 2003-05, Vol.84 (5), p.1173-1178
• Main Authors: Leisy, Douglas J, Lewis, Teresa D, Leong, Jo-Ann C, Rohrmann, George F
• Format: Article
• Language: English
• Subjects: Animals; Baculoviridae - genetics; Baculoviridae - pathogenicity; Baculovirus; beta-Galactosidase - genetics; beta-Galactosidase - metabolism; Carps; Cell Line; Cytomegalovirus; Fishes - virology; Freshwater; Genetic Vectors; Oncorhynchus keta; Oncorhynchus mykiss; Oncorhynchus tshawytscha; Salmon; Tilapia; Transduction, Genetic; Vesicular stomatitis virus
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/vir.0.18861-0

1 Department of Microbiology, Oregon State University, 220 Nash Hall, Corvallis, OR 97331-3804, USA 2 Hawaii Institute of Marine Biology, Kaneohe, HI 96744, USA Correspondence Douglas Leisy leisyd{at}onid.orst.edu Five fish cell lines were tested for their ability to be transduced by Ac-CA lacZ , a recombinant baculovirus that is capable of expressing a -galactosidase reporter gene from the CAG promoter (consisting of a cytomegalovirus enhancer element, a chicken actin promoter and rabbit -globin termination sequences). TO ( Tilapia ovary), EPC (carp), CHH-1 (Chum salmon heart fibroblast) and CHSE-214 (chinook salmon embryo) cells were transducible, as demonstrated by an in situ -galactosidase assay, whereas RTG-2 (rainbow trout gonad) cells were not. The EPC cell line was used for more detailed studies on baculovirus transduction. The transduction frequency was found to be higher at 28 °C than at 21 °C. Addition of the histone deacetylase inhibitor sodium butyrate increased the number of blue cells detected 5- to 7-fold. The m.o.i. was positively correlated with transduction frequency, although the relationship did not appear to be strictly linear, as has been observed with mammalian cells. The temperature at which baculoviruses were adsorbed to EPC cells did not affect levels of -galactosidase expression. We also examined expression levels of -galactosidase in EPC cells after infection with a baculovirus construct that overexpresses the vesicular stomatitis virus G protein and displays it on the virion surface. Expression levels with this virus were approximately 15-fold higher than were observed with Ac-CA lacZ .