Using a Sonication-Assisted Agrobacterium Transformation System to Produce Mouse Granulocyte Macrophage Colony-Stimulating Factor in Rice Suspension Cells

• Main Authors: HSIEH,SHIH HAN, 謝世涵
• Other Authors: LIU,YU KUO
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/76685631137392360059

碩士 === 長庚大學 === 生化與生醫工程研究所 === 95 === Transgenic plants offer several advantages such as safety, cheaper cost and scalable production, as vectors for expression of the recombinant proteins. GM-CSF is a cytokine that functions as a white blood cell growth factor. GM-CSF stimulates stem cells to produce granulocytes and monocytes. In this study, we attempt to produce transgenic rice expressing mouse GM-CSF. In the expressing cassette, we used an constitutive-expression modified GluB-1 promoter and fused a signal peptide gene to develop a secretary system. Rice cells were transformed by using an Agrobacterium-mediated system. We select and culture the transformed cell lines in the medium including hygromycin. The high expression cell line could be selective and be proceeded to the protein analysis. We utilized SAAT (sonication-assisted Agrobacterium transformation) method to transform rice suspension cells directly. In this study, the productivity of mGM-CSF could reach to 1.523 ng/L under sucrose provision medium for 5 days and to 0.03 ng/L under sugar starvation medium for 3 days.