본 발명은 목적 단백질을 발현하는 식물세포, 이의 제조방법 및 이를 이용한 목적 단백질의 생산방법에 대한 것이다. 상기 식물세포에는 목적 단백질을 코딩하는 유전자를 함유하는 재조합 벡터가 도입되어 있고, 상기 식물세포는 식물 형성층 유래 줄기세포(CMC) 또는 캘러스를 포함한다. 이때, 상기 식물 형성층 유래 줄기세포(CMC)는 식물로부터 분리된 선천적으로 미분화된 세포들을 함유하는 식물 유래 세포주로써, 상기 세포주는 식물의 형성층 조직으로부터 분리되고, 캘러스로의 탈분화과정을 거치치 아니한 분열조직적 연속성을 가지는 것을 특징으로 한다. 본 발명에 따른 목적 단백질 발현 시스템은 종래 식물세포 배양의 문제점을 해소하며, 획기적인 형질전환율로 인하여 바이오의약 단백질을 포함하는 목적 단백질을 대량생산 할 수 있게 하여 식물 유래 단백질 제품 등의 바이오의약품의 상업화를 가능하게 하는 바, 유용하다.

Virus like particles comprising a fusion protein and substantially free of nucleic acid, wherein the fusion protein comprises a plant viral coat protein and a target protein, are provided. Immunogenic compositions comprising the virus like particles can be administered to subjects to induce protective immune responses in the subjects. Methods of producing the virus like particles are also provided.

Provided is a method for producing a target protein via cultivation of transgenic plant cells comprising a promoter capable of expressing the protein under sugar-free conditions or in response to the depletion of sugar and a gene encoding the target protein, without exchange of a cell growth medium with a sugar-depleted medium. The method comprises 1 ) culturing transgenic plant cells in a sugar-rich medium to grow plant cells; and 2) culturing the transgenic plant cells with addition of an amino acid mixture to the culture of Step 1 without exchange of a cell growth medium with a sugar-depleted medium, thereby expressing a target protein. The method of the present invention enables commercial-scale production of recombinant proteins via establishment of optimized culture conditions by addition of an amino acid mixture to induce protein expression without exchange of a cell growth medium with a sugar- free medium, and addition of pyruvic acid during the induction period of protein expression to thereby enhance the production yield of target proteins.

Plant genes regulated by transcription factors that control the gene network response to an environmental perturbation or signal are described. This class of genes responds to the perturbation of a transcription factor and the signal it transduces, but surprisingly, without stable binding of the transcription factor. These genes represent members of the "dark matter" of metabolic regulatory circuits. The invention involves the transgenic manipulation of these "response genes" and/or the genes encoding their regulatory transcription factors in plants so that their respective gene products are either overexpressed or underexpressed in the plant in order to confer a desired phenotype. The invention also relates to a rapid technique named "TARGET" ( T ransient A ssay R eporting G enome-wide E ffects of T ranscription factors) for determining such "response genes" and their transcription factors by perturbation of the expression of the transcription factors of interest in protoplasts of any plant species.

　植物細胞において、相同組換えによりゲノムＤＮＡ内に挿入され、その両末端にヌクレアーゼ認識部位が付加されているマーカー遺伝子を、対応するヌクレアーゼにより除去することができ、さらにヌクレアーゼ認識部位に隣接する少なくとも３０ヌクレオチドの配列を一致させることにより、該認識部位も痕跡残すことなく除去できることを見出した。そして、相同組換えにより植物細胞のゲノム上の標的ＤＮＡに変異を導入する方法において、該変異が導入された植物細胞の選択を、マーカー遺伝子の発現を指標として安定的に行うことが可能となり、さらに選択された細胞において、マーカー遺伝子等の不要な配列を除去し、標的ＤＮＡに必要な変異のみを導入することが可能なった。

[00408] Plant genes regulated by transcription factors that control the gene network response to an environmental perturbation or signal are described. This class of genes responds to the perturbation of a transcription factor and the signal it transduces, but surprisingly, without stable binding of the transcription factor. These genes represent members of the "dark matter" of metabolic regulatory circuits. [00409] The invention involves the transgenic manipulation of these "response genes" and/or the genes encoding their regulatory transcription factors in plants so that their respective gene products are either overexpressed or underexpressed in the plant in order to confer a desired phenotype. [00410] The invention also relates to a rapid technique named "TARGET" ( T ransient A ssay R eporting G enome-wide E ffects of T ranscription factors) for determining such "response genes" and their transcription factors by perturbation of the expression of the transcription factors of interest in protoplasts of any plant species.

본 발명은 번역 효율을 증진시키기 위하여 합성한 DNA 단편 및 이를 포함하는 재조합 벡터에 관한 것으로서, 보다 구체적으로 목적 단백질의 번역 능력을 향상시킬 수 있는 서열번호 1로 표시되는 21개의 염기서열로 구성된 DNA 단편 및 상기 DNA 단편을 포함하는 재조합 벡터에 관한 것이다. 본 발명에 따른 DNA 단편은 목적 단백질의 상류에 첨가되어 재조합 벡터로 제작됨으로써, 상기 재조합 벡터로 형질 전환된 식물체는 목적 단백질의 번역을 향상시킬 수 있고, 식물체 내의 특정 장소로 타겟팅을 유도하여 목적 단백질이 안정하게 축적될 수 있도록 하며, 이로 인해 식물체로부터 목적 단백질을 대량으로 생산할 수 있는 효과가 있다.