Combined hydrophobic-metal binding fusion tags for applications in aqueous two-phase partitioning |
| |
Authors: | Bernaudat Florent Bülow Leif |
| |
Institution: | Department of Pure and Applied Biochemistry, Centre for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, 221 00 Lund, Sweden. florent.bernaudat@tbiokem.lth.se |
| |
Abstract: | In this work, we studied the influence of fusion affinity tags containing both hydrophobic and histidines residues on the partitioning of the green fluorescent protein, GFPuv, in aqueous two-phase system. The tags were fused to the N-terminal of GFPuv and tested by immobilized metal affinity partitioning, in a PEG/salt system. The presence of both types of residues in the tag increased the partitioning greatly. Particularly, four engineered tags (H6, FH6, WH6, and YH6) containing a hexa-histidine sequence as well as different hydrophobic residues, all increased partitioning more than twice, reaching K values around 20, as compared to another construct (His6-GFP) containing an isolated hexa-histidine sequence. YH6, also proved be beneficial for protein expression. |
| |
Keywords: | Peptide library Escherichia coli Screening Immobilized metal affinity chromatography Affinity gel electrophoresis Green fluorescent protein Aqueous two-phase system Surface plasmon resonance |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|