Tobacco BY2 cells containing a tomato mosaic virus vector for protein production
|Posted date||Jun 22, 2011|
|Country||United States of America|
|Date of filing||Oct 1, 2004|
|Gazette Date||Jan 15, 2009|
|Gazette Date||Aug 13, 2013|
|International application number||JP2004014487|
|International publication number||WO2005033306|
|Date of international filing||Oct 1, 2004|
|Date of international publication||Apr 14, 2005|
|Title||Tobacco BY2 cells containing a tomato mosaic virus vector for protein production|
An expression vector is constructed by transferring recombinant tomato mosaic virus (ToMV) cDNA, in which a coat protein gene of ToMV having a suppressor against a virus resistant reaction has been substituted by a GFP gene, into the downstream of a promoter capable of inducing steroid hormone-dependent transcription.
In a transformed tobacco BY-2 cell obtained by transferring the above expression vector into a tobacco BY-2 cells, steroid hormone-dependent transcription is induced, thereby enabling the amplification of mRNA of the GFP gene and induction of the expression of GFP.
|Scope of claims||
1. A process for producing a tobacco BY2 cell for protein production, comprising: a first transforming step of transfecting tobacco BY2 cells with a plasmid, the plasmid comprising a transcription factor-expressing DNA comprising a coding gene of a transcription factor, wherein the transcription factor is LexA-VP16-hER which is activated by estrogen; and
a promoter for expressing the transcription factor;
a screening step of screening tobacco BY2 cells, obtained in the first transforming step, for an individual tobacco BY2 cell expressing the transcription factor; and
a second transforming step of transfecting the individual tobacco BY2 cell, obtained in the screening step, with a plasmid, the plasmid comprising a protein-expressing DNA comprising cDNA of a RNA virus vector having incorporated therein a gene encoding a target protein, wherein the RNA virus vector is a tomato mosaic virus vector;
an inducible promoter which is OLexA-46 located upstream of the cDNA of the RNA virus vector, wherein the virus vector cDNA is transcribed under the control of the inducible promoter; and
a ribozyme sequence of satellite tobacco ringspot virus directly ligated to the 3' end of the RNA virus vector cDNA with no sequence ligated therebetween.
2. A process for producing a tobacco BY2 cell for protein production as set forth in claim 1, wherein the gene encoding the target protein replaces a viral coat protein gene.
3. A process for producing a tobacco BY2 cell for protein production as set forth in claim 1, wherein the transcription factor-expressing DNA and the protein-expressing DNA are transfected by an Agrobacterium method.
4. A plant culture tobacco BY2 cell for protein production, which is produced by the process for producing a tobacco BY2 cell for protein production as set forth in claim 1.
5. A protein producing process comprising, culturing the tobacco BY2 cell of claim 4 in the presence of estrogen.
|IPC(International Patent Classification)||
|Reference ( R and D project )||CREST Plants Function and Their Control AREA|
Contact Information for " Tobacco BY2 cells containing a tomato mosaic virus vector for protein production "
- Japan Science and Technology Agency Department of Intellectual Property Management
- URL: http://www.jst.go.jp/chizai/
- Address: 5-3, Yonbancho, Chiyoda-ku, Tokyo, Japan , 102-8666
- Fax: 81-3-5214-8476