Transduction of cultured fish cells with recombinant baculoviruses.
Five fish cell lines were tested for their ability to be transduced by Ac-CAlacZ, a recombinant baculovirus that is capable of expressing a beta-galactosidase reporter gene from the CAG promoter (consisting of a cytomegalovirus enhancer element, a chicken actin promoter and rabbit beta-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 beta-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 degrees C than at 21 degrees 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 beta-galactosidase expression. We also examined expression levels of beta-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-CAlacZ.
Leisy
Douglas J
DJ
Department of Microbiology, Oregon State University, 220 Nash Hall, Corvallis, OR 97331-3804, USA.
Lewis
Teresa D
TD
Hawaii Institute of Marine Biology, Kaneohe, HI 96744, USA.
Leong
Jo-Ann C
JC
Hawaii Institute of Marine Biology, Kaneohe, HI 96744, USA.|Department of Microbiology, Oregon State University, 220 Nash Hall, Corvallis, OR 97331-3804, USA.
Rohrmann
George F
GF
Department of Microbiology, Oregon State University, 220 Nash Hall, Corvallis, OR 97331-3804, USA.
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000472:pathogenicity*; D002347:Carps; D002460:Cell Line; D005399:Fishes / Q000821:virology*; D005822:Genetic Vectors; D017686:Oncorhynchus mykiss; D012474:Salmon; D017210:Tilapia; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
D001616:beta-Galactosidase