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The Expression of M-CSF Soluble Receptor in Tobacco Plants and Its Anti-M-CSF Effect
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Abstract:
Objective:To effectively express human M-CSF soluble receptor in tobacco plants and study its anti-M-CSF activity. Methods:The plant expression plasmid expressing recombinant human M-CSF soluble receptor with C-terminal His-tag and endoplasmic reticulum retention sequence was constructed. Then the target gene was introduced into tobacco genome by agrobacterium tumefaciens-mediated transformation. PCR and Western blot were used to select the transgenic tobacco plants. ELISA was used to evaluate the expression level while colony forming assay was used to test its biological activity. Results:More than 50 transgenic tobacco plants were developed. PCR results showed the insertion of target gene into tobacco genome and Western blot results showed the expression of recombinant protein. The recombinant M-CSFsR was effectively expressed, however, the expression levels varied among different transgenic tobacco plants, with the maximum reaching 1.92% of total soluble protein in leaf tissues. The tobacco-derived M-CSFsR could inhibit the colony formation of J6-1 cells. Conclusion:Bioactive recombinant human M-CSFsR was expressed in transgenic tobacco plants at high level.
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