目的：在烟草中高效表达人M-CSF可溶性受体并研究其抗M-CSF活性。方法： 采用基因重组技术构建表达C端含组氨酸标签和内质网滞留信号的M-CSF可溶性受体植物表达载体，通过根瘤土壤杆菌转化烟草，获得转基因植株。PCR、蛋白印迹实验鉴定转基因植株，ELISA方法测定重组蛋白表达水平，集落形成实验测定重组蛋白活性。结果：获得50余株转基因烟草植株，PCR证实目的基因已整合到烟草基因组中，蛋白印迹实验证实转基因植株表达目的蛋白；M-CSFsR在烟草中获得高效表达，但不同转基因植株的表达水平存在差异，其中表达水平最高的一株M-CSFsR占叶片可溶性蛋白的1.92%；集落形成实验证明M-CSFsR可以抑制J6-1细胞的集落形成。结论：在烟草中高效表达有生物学活性的重组人M-CSFsR。

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.

天津市科技发展计划项目（003119311）; 国家自然科学基金（30470897）