Effect of location of the His-tag on the production of soluble and functional Buthus martensii Karsch insect toxin

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• 作者：Cheng-Gang Xu ; Xiao-Jun Fan ; Yue-Jun Fu ; Ai-Hua Liang
• 关键词：Cysteine-rich proteins ; Buthus martensii Karsch insect toxin (BmK IT) ; Oxidative refolding ; Thermostability ; Circular dichroism spectroscopy
• 刊名：Protein Expression and Purification
• 出版年：2008
• 期刊代码：184_10465928
• 类别：bio
• 出版时间：May 2008
• 卷：59
• 期：1
• 页码：103-109
• 文件大小：502 K

摘要

The low yield and poor folding efficiency in vivo of soluble and active recombinant cysteine-rich proteins expressed in Escherichia coli are a major challenge for large-scale protein production and purification. Expression vectors containing Buthus martensii Karsch insect toxin (BmK IT) fused to the C terminus of the intein Ssp DnaB were constructed in an attempt to overcome this problem. Following purification and intein self-cleavage, the fusion protein His₆-intein-IT produced insoluble BmK IT, while intein-IT-His₆ generated soluble and properly folded BmK IT. This result indicated that the positioning of the His₆ tag has a key role in the production of soluble and functional BmK IT.