实现人源FGF-21高效可溶性表达的两种策略

李娟; 刘雯; 肖磊; 劳勋; 赵丽芬; 黄静; 吴自荣

实现人源FGF-21高效可溶性表达的两种策略

1.
华东师范大学生命科学学院,上海 200062

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出版历程
- 收稿日期: 2011-09-01
- 修回日期: 2011-12-01
- 刊出日期: 2012-11-25

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

1.
School of Life Sciences, East China Normal University, Shanghai 200062， China

摘要

摘要: 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〖WTBX〗OD〖WTBZ〗600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(〖WTBX〗P〖WTBZ〗0.01).
- 成纤维生长因子21 /
- 分子伴侣 /
- 乳糖自诱导
Abstract: 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 〖WTBX〗OD600 〖WTBZ〗 was 12.4; thus relative yield was 5.5 times higher than IPTG induction. The biological activities of recombinant FGF-21 were studied 〖WTBX〗in vivo〖WTBZ〗 in healthy C57BL/6 mice. It showed that recombinant FGF-21 can lower blood glucose obviously in mice (〖WTBX〗P〖WTBZ〗 0.05 or 〖WTBX〗P 〖WTBZ〗0.01). The recombinant FGF-21 treatment also led to a significant decrease in concentration of triglyceride in C57BL/6 mice (〖WTBX〗P〖WTBZ〗 0.01).
- fibroblast growth factor 21 /
- molecular chaperone /
- lactose auto-induction

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参考文献(1)

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