Production of mouse granulocyte-macrophage colony-stimulating factor by gateway technology and transgenic rice cell culture

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题名:	Production of mouse granulocyte-macrophage colony-stimulating factor by gateway technology and transgenic rice cell culture
作者:	陸重安;Liu, Yu-Kuo;Huang, Li-Fen;Ho, Shin-Lon;Liao, Chun-Yu;Liu, Hsin-Yi;Lai, Ying-Hui;Yu, Su-May;Lu, Chung-An
贡献者:	生醫理工學院生命科學系
关键词:	Agrobacterium - genetics;Animals;bioreactor;Bioreactors;Biotechnology - methods;Cell culture;Cell Culture Techniques;Cloning;Crop production;Culture Media - chemistry;DNA, Bacterial;E coli;Escherichia coli;Escherichia coli - genetics;Gene expression;Genetic Vectors;Granulocyte-Macrophage Colony-Stimulating Factor - chemistry;Granulocyte-Macrophage Colony-Stimulating Factor - genetics;Granulocyte-Macrophage Colony-Stimulating Factor - metabolism;Leukemia;mGM-CSF;Mice;Molecular Weight;Oryza - genetics;Oryza - metabolism;Oryza sativa;Plants, Genetically Modified;Promoter Regions, Genetic;Proteins;recombinant protein production;Recombinant Proteins - chemistry;Recombinant Proteins - genetics;Recombinant Proteins - metabolism;rice suspension cell;Rodents;Signal transduction;sugar-depletion induced promoter;Transgenic plants
日期:	2012-05-01
上传时间:	2026-04-23 11:13:38 (UTC+8)
出版者:	Wiley-VCH Verlag;Hoboken: Wiley Subscription Services, Inc., A Wiley Company
摘要:	摘要： 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. Biotechnol. Bioeng. 2012; 109:1239–1247. © 2011 Wiley Periodicals, Inc. The authors constructed a Gateway‐compatible binary T‐DNA destination vector for the easy establishment of a production platform for recombinant proteins in rice suspension cells. Here, the 2‐L bioreactor was used for the production of recombinant mouse granulocyte‐macrophage colony‐stimulating factor (mGM‐CSF), a cytokine that functions in the growth of white blood cells, in cultured rice suspension cells. The recombinant mGM‐CSF was bioactive and could stimulate the proliferation of a murine myeloblastic leukemia cell. 其他題名： Biotechnol. Bioeng 出版者： Hoboken: Wiley Subscription Services, Inc., A Wiley Company 出版日期： 2012-05 出處： Biotechnology and bioengineering, 2012-05, Vol.109 (5), p.1239-1247 資源來源： Wiley Online Library All Journals 版權： Copyright © 2011 Wiley Periodicals, Inc. 版權： Copyright John Wiley and Sons, Limited May 2012 識別號： ISSN: 0006-3592 識別號： ISSN: 1097-0290 識別號： EISSN: 1097-0290 識別號： DOI: 10.1002/bit.24394 識別號： PMID: 22125231 識別號： CODEN: BIBIAU
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