Effect of location of the His-tag on the production of soluble and functional Buthus martensii Karsc
Effect of location of the His-tag on the production of soluble and functional Buthus martensii Karsch insect toxin.
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 His(6)-intein-IT produced insoluble BmK IT, while intein-IT-His(6) generated soluble and properly folded BmK IT. This result indicated that the positioning of the His(6) tag has a key role in the production of soluble and functional BmK IT.
Xu CG, Fan XJ, Fu YJ, Liang AH.
Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Wucheng Road 36, Taiyuan, Shanxi 030006, PR China.