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RANKL/RANK/OPG轴在骨代谢过程中起到中心调节作用,也是近年来骨相关疾病治疗研究的热点之一。RANKL蛋白在RANKL/RANK/OPG轴信号传递过程中起到关键作用,在骨代谢相关实验研究中用途广泛。但是,使用大肠杆菌Escherichia coli可溶表达重组人源RANKL蛋白(hRANKL)时产量远低于鼠源RANKL(mRANKL)。本研究通过将LB培养基pH值调整并稳定在7.5、降低诱导表达温度至16℃并优化细菌裂解条件,成功地将可溶hRANKL产量增加到了对照组的5-12倍。该方法有效提高了hRANKL在大肠杆菌中可溶表达的产量,同时也是研究重组蛋白在大肠杆菌内的可溶表达策略的有益尝试。
RANKL / RANK / OPG axis plays a central role in the process of bone metabolism, and is also one of the hot spots in the research of the treatment of bone-related diseases in recent years. RANKL protein plays a key role in RANKL / RANK / OPG axis signal transduction, and is widely used in the experimental research of bone metabolism. However, the yield of recombinant human RANKL protein (hRANKL) soluble in E. coli Escherichia coli was much lower than that of murine RANKL (mRANKL). In this study, the yield of soluble hRANKL was successfully increased to 5-12 times of that of the control by adjusting and stabilizing the LB medium pH at 7.5, reducing the induction temperature to 16 ° C and optimizing bacterial lysis conditions. The method effectively increases the yield of soluble expression of hRANKL in E. coli and is also a beneficial attempt to study the soluble expression strategy of the recombinant protein in E. coli.