Part:BBa_K2306005

Cytosolic-abundant heat soluble protein 106094 (CAHS 106094)

This Biobrick features the gene sequence that encodes for the production of the tardigrade intrinsically disordered protein (TDP) "CAHS 106094". This is one of the several heat soluble proteins found in the tardigrades responsible for the ability of these microscopic animals to survive the harshest conditions. Upon desiccation, tardigrade proteins form a glass matrix capable of protecting other fragile biological components. Therefore, this tardigrade protein can be combined with other perishable biological material (e.g. proteins) to increase its stability against dehydration or even enable the possibility of stabilizing material with just a previous drying step. This BioBrick is featured in the composite part BBa_K2306010 combined with a T7 promoter BBa_R0010, a ribosome binding site BBa_k2306014 and a double terminator BBa_B0015 to control its expression. Further information about our project can be found on our [http://2017.igem.org/Team:TUDelft/Results#TDP results page].

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Contribution of TUM-Straubing 2024

BBa_K2306005 is known for its protective role in both Hypsibius dujardini [1] and living E. coli cells. In our experiments, BBa_K2306005 demonstrated protective capabilities of cell free expression systems when produced internally, and externally supplemented proteins exhibited diminished protection. This reduction is possibly due to an inhibitory effect of the storage buffer, which contains 50 mM NaCl. Surprisingly, upon further validation, BBa_K2306005 performed significantly better than in earlier assessments, indicating variability between experimental batches and handling during the desiccation process.

References

[1] Boothby, T. C., Tapia, H., Brozena, A. H., Piszkiewicz, S., Smith, A. E., Giovannini, I., Rebecchi, L., Pielak, G. J., Koshland, D., & Goldstein, B. (2017). Tardigrades Use Intrinsically Disordered Proteins to Survive Desiccation. Molecular Cell, 65(6), 975. https://doi.org/10.1016/j.molcel.2017.02.018