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Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants
Authors:Zheng Guo-Guang  Yang Ying-Hua  Rao Qing  Lin Yong-Min  Zhang Bin  Wu Ke-Fu
Institution:State Key Laboratory for Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, PR China. zhengggtj@sohu.com
Abstract:The cDNA encoding N-terminal three immunoglobin-like domains of human M-CSFR was linked to His-tag and endoplasmic reticulum retention sequence (KDEL) before being inserted into the genome of tobacco plant, Nicotiana tabacum cv. NC-89, by Agrobacterium tumefaciens-mediated transformation. The insertion and expression of target gene were confirmed by PCR, ELISA, and Western blot. The recombinant M-CSFsR reached a maximum expression level of 1.92% of total soluble protein in transgenic tobacco plant leaf tissues. The recombinant M-CSFsR could be purified through a one-step IMAC process and its bioactivity was confirmed by the inhibition of colony formation of J6-1 cells. The results suggested that we successfully expressed a high level of bioactive human M-CSFsR in tobacco plants.
Keywords:Bioactive  Macrophage colony-stimulating factor  Soluble receptor  Tobacco  Transgenic plants
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