Design and establishment of a vector system that enables production of multifusion proteins and easy purification by a two-step affinity chromatography approach.
Journal of Microbiological Methods（2014）
Abstract
The LE (LguI/Eco81I)-cloning procedure allows a step-wise, directional fusion of multiple DNA-fragments into a vector by utilizing two restriction enzymes generating identical non-palindromic overhangs. This strategy was applied to produce heat-stable cellulase-fusion proteins containing up to five single moieties. Terminal affinity tags enable efficient purification using a simple two-step approach.
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Key words
Gene fusion,Multiple fusion approach,Type IIS restriction enzyme,Two-step affinity chromatography,Heat-stable cellulases
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