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

外国特許コード F110003325
整理番号 A181-07WO
掲載日 2011年6月22日
出願国 アメリカ合衆国
出願番号 57411204
公報番号 20090017490
公報番号 8507220
出願日 平成16年10月1日(2004.10.1)
公報発行日 平成21年1月15日(2009.1.15)
公報発行日 平成25年8月13日(2013.8.13)
国際出願番号 JP2004014487
国際公開番号 WO2005033306
国際出願日 平成16年10月1日(2004.10.1)
国際公開日 平成17年4月14日(2005.4.14)
優先権データ
  • 特願2003-343747 (2003.10.1) JP
  • 特願2003-350091 (2003.10.8) JP
  • 特願2004-056912 (2004.3.1) JP
  • 2004WO-JP14487 (2004.10.1) WO
発明の名称 (英語) Tobacco BY2 cells containing a tomato mosaic virus vector for protein production
発明の概要(英語) (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.
特許請求の範囲(英語) [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.
  • 発明者/出願人(英語)
  • MORI MASASHI
  • DOHI KOJI
  • ISHIKAWA MASAYUKI
  • MESHI TETSUO
  • NISHIKIORI MASAKI
  • TAMAI ATSUSHI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
国際特許分類(IPC)
米国特許分類/主・副
  • C12N015/82B24
参考情報 (研究プロジェクト等) CREST Plants Function and Their Control AREA
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