| 大肠杆菌表达的vMIP-Ⅱ包涵体的纯化与复性研究 |
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| 引用本文： | 张少恩 孙晗笑 张光 杨清玲 沙文琼 刘俊云 龚莉桂. 大肠杆菌表达的vMIP-Ⅱ包涵体的纯化与复性研究[J]. 中国生物制品学杂志, 2005, 18(3): 247-251 |
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| 作者姓名： | 张少恩 孙晗笑 张光 杨清玲 沙文琼 刘俊云 龚莉桂 |
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| 作者单位： | 暨南大学药学院基因组药物研究所 广州510632(张少恩，孙晗笑，张光，杨清玲，沙文琼，刘俊云)，暨南大学药学院基因组药物研究所 广州510632(龚莉桂) |
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| 基金项目： | 国家“八六三”计划项目(No.2003AA219060)，国家自然科学基金(No.30170881)，广东省重点科技攻关计划项目(No.2002A1090211). |
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| 摘 要： | 目的纯化、复性大肠杆菌表达的vMIPⅡ包涵体蛋白。方法用8molL尿素缓冲液变性溶解vMIPⅡ包涵体,然后加入十二烷基磺酸钠(SDS)进一步促溶,利用金属亲和层析和分子筛层析纯化,再利用柱层析去除SDS,纯化后透析复性;通过荧光和单核淋巴细胞趋化抑制实验检测复性蛋白的结构和抑制单核淋巴细胞趋化的活性。结果重组vMIPⅡ纯化和复性后,仍具有完整的高级结构和抑制单核细胞趋化的活性。结论已获得了重组vMIPⅡ活性蛋白,为进一步研究应用奠定基础。
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| 关 键 词： | vMIP-Ⅱ包涵体 十二烷基磺酸钠 环氧糊精 |
| 修稿时间： | 2004-07-23 |
Purification and Renaturation of vMIP- Ⅱ Inclusion Body Expressed in E. coli |
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| ZHANG Shao-en,SUN Han-xiao,ZHANG Guang,et al. Purification and Renaturation of vMIP- Ⅱ Inclusion Body Expressed in E. coli[J]. Chinese Journal of Bilogicals, 2005, 18(3): 247-251 |
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| Authors: | ZHANG Shao-en SUN Han-xiao ZHANG Guang et al |
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| Abstract: | Objective To purify and re-naturalize the vMIP-Ⅱ inclusion body protein expressed in E.coli.Methods The vMIP-Ⅱ inclusion body expressed in E.coli was dissolved with 8 mol/L urea,then further dissolved with SDS and purified by metal affinity chromatography and molecular sieve chromatography.Revmove the SDS bound to protein molecules by affinity column chromatography and re-naturalize the vMIP-Ⅱ inclusion body by dialysis.Examine the structure of renaturalized protein by fluorometric assay and determine its activity by chemotaxis-inhibiting test of mononuclear lymphocytes.Results The recombinant vMIP-Ⅱ with intact higher-order structure was purified,re-naturalized,and showed activity in inhibiting the chemotaxis of mononuclear lymphocyte.Conclusion The recombinant vMIP-Ⅱ protein with activity was obtained.It laid a foundation of further application of the protein. |
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| Keywords: | vMIP-Ⅱ inclusion body SDS Cyclodextrins |
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