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学科主题: 生命有机化学
题名: Highly Efficient Production of Soluble Proteins from Insoluble Inclusion Bodies by a Two-Step-Denaturing and Refolding Method
其他题名: 通过两步变复性实现了从难溶的包涵体中得到可溶性蛋白的高效表达
作者: YANG ZHONG ; ZHANG LINLIN ; Zhang Y(张岩) ; ZHANG TING ; FENG YANYE ; Lu XX(卢秀秀) ; Lan WX(蓝文贤) ; Wang Jufang ; Wu HM(吴厚铭) ; Cao CY(曹春阳) ; WANG XIAONING
通讯作者: 曹春阳 ; WANG XIAONING
刊名: PLoS One
发表日期: 2011
卷: 6, 期:7, 页:e22981
收录类别: SCI
部门归属: 复旦大学; 中科院上海有机化学研究所; 华东理工大学; 华南理工大学
英文摘要: The production of recombinant proteins in a large scale is important for protein functional and structural studies, particularly by using Escherichia coli over-expression systems; however, approximate 70% of recombinant proteins are over-expressed as insoluble inclusion bodies. Here we presented an efficient method for generating soluble proteins from inclusion bodies by using two steps of denaturation and one step of refolding. We first demonstrated the advantages of this method over a conventional procedure with one denaturation step and one refolding step using three proteins with different folding properties. The refolded proteins were found to be active using in vitro tests and a bioassay. We then tested the general applicability of this method by analyzing 88 proteins from human and other organisms, all of which were expressed as inclusion bodies. We found that about 76% of these proteins were refolded with an average of >75% yield of soluble proteins. This "two-step-denaturing and refolding" (2DR) method is simple, highly efficient and generally applicable; it can be utilized to obtain active recombinant proteins for both basic research and industrial purposes.
语种: 英语
相关网址: 查看原文
WOS记录号: WOS:000293286500062
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内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/17405
Appears in Collections:上海有机化学研究所_期刊论文

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Recommended Citation:
YANG ZHONG,ZHANG LINLIN,Zhang Y,et al. Highly Efficient Production of Soluble Proteins from Insoluble Inclusion Bodies by a Two-Step-Denaturing and Refolding Method[J]. PLoS One,2011,6(7):e22981.
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