Inducible virus-mediated expression of a foreign protein in suspension-cultured plant cells.
Although suspension-cultured plant cells have many potential merits as sources of useful proteins, the lack of an efficient expression system has prevented using this approach. In this study, we established an inducible tomato mosaic virus (ToMV) infection system in tobacco BY-2 suspension-cultured cells to inducibly and efficiently produce a foreign protein. In this system, a modified ToMV encoding a foreign protein as replacement of the coat protein is expressed from stably transformed cDNA under the control of an estrogen-inducible promoter in transgenic BY-2 cells. Estrogen added to the culture activates an estrogen-inducible transactivator expressed constitutively from the transgene and induces transcription and replication of viral RNA. In our experiments, accumulation of viral RNA and expression of green fluorescent protein (GFP) encoded in the virus were observed within 24 h after induction. The amount of GFP reached approximately 10% of total soluble protein 4 d after induction. In contrast, neither viral RNA nor GFP were detected in uninduced cells. The inducible virus infection system established here should be utilized not only for the expression of foreign proteins, but also for investigations into the viral replication process in cultured plant cells.
Dohi
K
K
Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Ishikawa, Japan.
Nishikiori
M
M
Tamai
A
A
Ishikawa
M
M
Meshi
T
T
Mori
M
M
D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D004958:Estradiol / Q000494:pharmacology; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012367:RNA, Viral / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014026:Nicotiana / Q000166:cytology / Q000235:genetics / Q000821:virology*; D017898:Tobamovirus / Q000235:genetics*; D015534:Trans-Activators / Q000187:drug effects / Q000235:genetics; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D019076:Transgenes; D014779:Virus Replication
D036022:Capsid Proteins; D012367:RNA, Viral; D011994:Recombinant Proteins; D015534:Trans-Activators; D049452:Green Fluorescent Proteins; D004958:Estradiol