Abstract
The aim of this work was to construct short analogues of the repetitive water-binding domain of the Pseudomonas syringae ice nucleation protein, InaZ. Structural analysis of these analogues might provide data pertaining to the protein-water contacts that underlie ice nucleation. An artificial gene coding for a 48-mer repeat sequence from InaZ was synthesized from four oligodeoxyribonucleotides and ligated into the expression vector, pGEX2T. The recombinant vector was cloned in Escherichia coli and a glutathione S-transferase fusion protein obtained. This fusion protein displayed a low level of ice-nucleating activity when tested by a droplet freezing assay. The fusion protein could be cleaved with thrombin, providing a means for future recovery of the 48-mer peptide in amounts suitable for structural analysis by nuclear magnetic resonance spectroscopy.
| Original language | English |
|---|---|
| Pages (from-to) | 73-78 |
| Number of pages | 6 |
| Journal | Gene |
| Volume | 142 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 13 May 1994 |
Keywords
- gene synthesis
- fusion protein
- protein purification
- synthetic oligodeoxyribonucleotides
- Western analysis
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