Design and establishment of a vector system that enables production of multifusion proteins and easy purification by a two-step affinity chromatography approach

作者：Marquardt Tabea; von der Heyde Amelie; Elleuche Skander^*

来源：Journal of Microbiological Methods, 2014, 105: 47-50.

DOI：10.1016/j.mimet.2014.07.008

• Gene fusion
• Multiple fusion approach
• Type IIS restriction enzyme
• Two-step affinity chromatography
• Heat-stable cellulases

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.

• 出版日期2014-10