1. Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants.
- Author
-
Zheng GG, Yang YH, Rao Q, Lin YM, Zhang B, and Wu KF
- Subjects
- Cell Line, Humans, Plant Leaves genetics, Plant Leaves metabolism, Plants, Genetically Modified metabolism, Protein Structure, Tertiary genetics, Receptor, Macrophage Colony-Stimulating Factor genetics, Receptor, Macrophage Colony-Stimulating Factor isolation & purification, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins isolation & purification, Recombinant Fusion Proteins pharmacology, Nicotiana metabolism, Gene Expression, Plants, Genetically Modified genetics, Receptor, Macrophage Colony-Stimulating Factor biosynthesis, Recombinant Fusion Proteins biosynthesis, Nicotiana genetics
- 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.
- Published
- 2006
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