Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants |
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Authors: | Zheng Guo-Guang Yang Ying-Hua Rao Qing Lin Yong-Min Zhang Bin Wu Ke-Fu |
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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 |
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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. |
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Keywords: | Bioactive Macrophage colony-stimulating factor Soluble receptor Tobacco Transgenic plants |
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