Production of mouse granulocyte-macrophage colony-stimulating factor by gateway technology and transgenic rice cell culture |
| |
Authors: | Liu Yu-Kuo Huang Li-Fen Ho Shin-Lon Liao Chun-Yu Liu Hsin-Yi Lai Ying-Hui Yu Su-May Lu Chung-An |
| |
Institution: | Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Kwei-Shan, Taoyuan County, Taiwan, ROC. ykliu@mail.cgu.edu.tw |
| |
Abstract: | To establish a production platform for recombinant proteins in rice suspension cells, we first constructed a Gateway-compatible binary T-DNA destination vector. It provided a reliable and effective method for the rapid directional cloning of target genes into plant cells through Agrobacterium-mediated transformation. We used the approach to produce mouse granulocyte-macrophage colony-stimulating factor (mGM-CSF) in a rice suspension cell system. The promoter for the αAmy3 amylase gene, which is induced strongly by sugar depletion, drove the expression of mGM-CSF. The resulting recombinant protein was fused with the αAmy3 signal peptide and was secreted into the culture medium. The production of rice-derived mGM-CSF (rmGM-CSF) was scaled up successfully in a 2-L bioreactor, in which the highest yield of rmGM-CSF was 24.6 mg/L. Due to post-translational glycosylation, the molecular weight of rmGM-CSF was larger than that of recombinant mGM-CSF produced in Escherichia coli. The rmGM-CSF was bioactive and could stimulate the proliferation of a murine myeloblastic leukemia cell line, NSF-60. |
| |
Keywords: | bioreactor mGM‐CSF recombinant protein production rice suspension cell sugar‐depletion induced promoter |
本文献已被 PubMed 等数据库收录! |
|