实现人源FGF-21高效可溶性表达的两种策略

Two strategies for efficient expression of soluble recombinant human FGF-21 protein

采用分子伴侣协助蛋白表达的方法及乳糖自诱导的策略,实现人源FGF-21在大肠杆菌中的高效可溶性表达.结果显示,在分子伴侣Tig的协同表达下,融合蛋白TrxA-FGF-21可溶性达到92.4%.采用乳糖自诱导策略,融合蛋白TrxA-FGF-21表达量达到17.4%左右,菌体OD600达到12.4,可溶性TrxA-FGF-21相对产量是IPTG诱导发酵条件的5.5倍.该方法制备的重组FGF-21在C57BL/6小鼠体内具有降低血糖的生物学活性,甘油三酯水平与对照组相比也表现极显著差异(P<0.01).

The expression of recombinant soluble FGF-21 protein was achieved through transferring another plasmid pTf16 which expressed molecular chaperone Tig.On the other hand,the yield of FGF-21 was greatly improved with the method of lactose auto-induction.The result showed that the soluble TrxA-FGF-21 fusion protein reached 92.4% by molecular chaperone co-expression.With lactose auto-induction,the expression quantity of fusion protein was 17.4% of the total proteins,and the OD600 was 12.4;thus relative yield was 5.5 times higher than IPTG induction.The biological activities of recombinant FGF-21 were studied in vivo in healthy C57BL/6 mice.It showed that recombinant FGF-21 can lower blood glucose obviously in mice（P 0.05 or P 0.01）.The recombinant FGF-21 treatment also led to a significant decrease in concentration of triglyceride in C57BL/6 mice（P 0.01）.