Infection of cultured embryo cells of the pacific oyster, Crassostrea gigas, by pantropic retroviral vectors.
The inability to stably introduce and express foreign genes has hampered basic research in molluscan species. We cultured cells from dissociated embryos of the Pacific oyster, Crassostrea gigas, and infected these primary cultures with pantropic retroviral vectors containing the envelope glycoprotein of vesicular stomatitis virus. Luciferase transgene expression mediated by different heterologous promoters was demonstrated for at least 9 d after infection of the cells. Surprisingly, the promoter reproducibly mediating the highest level of luciferase expression was the retroviral promoter (U3 region of long terminal repeat) from the Moloney murine leukemia virus. The infection efficiency using a low multiplicity of infection (0.05) was estimated by quantitative polymerase chain reaction to be between 0.1-0.5%. This system will facilitate studies of gene expression and regulation and should be widely applicable to other molluscan species.
Boulo
V
V
IFREMER-CNRS, Universite Montpellier II, Defense et Resistance chez les Invertebres Marins, France. vboulo@ifremer.fr
Cadoret
J P
JP
Shike
H
H
Shimizu
C
C
Miyanohara
A
A
Burns
J C
JC
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D004625:Embryo, Nonmammalian / Q000166:cytology* / Q000821:virology; D017930:Genes, Reporter; D005822:Genetic Vectors; D006321:Heart / Q000196:embryology; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D009206:Myocardium / Q000166:cytology; D009997:Osmotic Pressure; D010125:Ostreidae / Q000166:cytology / Q000196:embryology* / Q000821:virology; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D012190:Retroviridae / Q000235:genetics* / Q000502:physiology
D011994:Recombinant Proteins; D008156:Luciferases