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Tobacco BY2 cells containing a tomato mosaic virus vector for protein production

Foreign code F110003325
File No. A181-07WO
Posted date Jun 22, 2011
Country United States of America
Application number 57411204
Gazette No. 20090017490
Gazette No. 8507220
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
Priority data
  • P2003-343747 (Oct 1, 2003) JP
  • P2003-350091 (Oct 8, 2003) JP
  • P2004-056912 (Mar 1, 2004) JP
  • 2004WO-JP14487 (Oct 1, 2004) WO
Title Tobacco BY2 cells containing a tomato mosaic virus vector for protein production
Abstract (US8507220)
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 [claim1]
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.
[claim2]
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.
[claim3]
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.
[claim4]
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.
[claim5]
5. A protein producing process comprising, culturing the tobacco BY2 cell of claim 4 in the presence of estrogen.
  • Inventor, and Inventor/Applicant
  • MORI MASASHI
  • DOHI KOJI
  • ISHIKAWA MASAYUKI
  • MESHI TETSUO
  • NISHIKIORI MASAKI
  • TAMAI ATSUSHI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) CREST Plants Function and Their Control AREA
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