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实现人源FGF-21高效可溶性表达的两种策略

, PP. 114-121

Keywords: 成纤维生长因子21,分子伴侣,乳糖自诱导

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Abstract:

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).

References

[1]  IZUMIYA Y, BINA H A, OUCHI N, et al. FGF-21 is an Akt-regulated myokine[J]. FEBS Lett, 2008, 582(17): 3805-3810.
[2]  NISHIMURA T, NAKATAKE Y, KONISHI M, et al. Identification of a nove FGF, FGF-21, preferentially expressed in the liver[J]. Biochim Biophys Acta, 2000, 1492(1): 203-206.
[3]  KHARTONENKOV A, SHIYANOVA T L, KOESTER A, et al. FGF-21 as a novel metabolic regulator[J]. J Clin Invest, 2005, 115(6): 1627-1635.
[4]  WENTE W, EFANOV A M, BRENNER M, et al. Fibroblast growth factor-21 improves pancreatic beta-cell function and survival by activation of extracellular signal-regulated kinase 1/2 and Akt signaling pathways[J]. Diabetes, 2006, 55(9): 2470-2478.
[5]  COSKUN T, BINA H A, SCHNEIDER M A, et al. Fibroblast growth factor 21 corrects obesity in mice[J]. Endocrinology, 2008, 149(12): 6018-6027.
[6]  姜媛媛,尹成凯,李晋南,等.SUMO 融合系统高效表达可溶性重组蛋白的研究[J].东北农业大学学报,2008, 39(10): 57-62.
[7]  崔硕硕,林学政,沈继红.分子伴侣共表达对低温脂肪酶Lip-837异源可溶性表达的影响[J].海洋科学进展,2011, 29(1):105-111.
[8]  ARAYO H, KAZUNORI H, NOBUTADA T, et al. Scalable purification and characterization of the extracellular domain of human autotaxin from prokaryotic cells[J]. Protein Expression and Purification, 2008, 59(1): 9-17.
[9]  顾娟,劳勋,金明飞,等.人胰高血糖素样肽-1突变体融合蛋白在大肠杆菌中的自诱导表达优化[J].微生物学通报,2010, 37(5): 726-731.
[10]  冯杉.T7表达系统及自诱导蛋白产出策略[J].北京教育学院学报:自然科学版,2009, 4(3): 10-15. 
[11]  COFFEY R J Jr, DERYNCK R, WILCOX J N, et al. Production and auto-induction of transforming growth factor-alpha in human keratinocyte[J]. Nature, 1987, 328(6133): 817-820.
[12]  STUDIER F W. Protein production by auto-induction in high density shaking cultures[J]. Protein Expr Purif, 2005, 41(1): 207-234.
[13]  LU Z S, CHEN W T, DING Y, et al. A novel method for high-level production of psychrophilic TAB5 alkaline phosphatase[J]. Protein Expression and Purification, 2010, 74(2): 217-222.
[14]  刘爽,胡宝成.原核系统可溶性表达策略[J].生物技术通讯,2005(2): 172-175.

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