Part:BBa_K1896018
INP_RC-mSA2 generator
This part contains the INPRC-mSA2 fusion protein controlled by a weak to medium constitutive promoter and RBS.
Usage and Biology
INPRC is a truncated Ice Nucleating Protein from which the N-terminal domain has been removed. The truncated INP is still capable of catalysing the formation of ice crystals, but is no longer localised to the outer cell membrane.[1] The mSA2 protein has been fused to INPRC for use in protein adhesion experiments performed by the UGent Belgium 2016 iGEM team. MSA2 is a monomeric variant of streptavidin that retains the biotin-binding capabilities and is better suited for protein fusions since it is less likely to cause aggregation.[2]
When crude cell lysate of a culture of E. coli expressing mGFPuv2 is added to supercooled water, no reaction takes place. When crude cell lysate containing INP_RC-mSA2 is added, ice is formed instantly. This demonstrates that the cytoplasmatic INP_RC fusion protein is still active as an ice nucleating protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 203
Illegal NgoMIV site found at 1715
Illegal NgoMIV site found at 1859
Illegal NgoMIV site found at 2123
Illegal NgoMIV site found at 2267
Illegal NgoMIV site found at 2315
Illegal NgoMIV site found at 2435
Illegal AgeI site found at 1379
Illegal AgeI site found at 3254
Illegal AgeI site found at 3314 - 1000COMPATIBLE WITH RFC[1000]
References
- Green, R. L., Corotto, L. V., & Warren, G. J. (1988). Deletion mutagenesis of the ice nucleation gene from Pseudomonas syringae S203. Molecular and General Genetics MGG, 215(1), 165-172.
- Mann, J. K., Demonte, D., Dundas, C. M., & Park, S. (2016). Cell labeling and proximity dependent biotinylation with engineered monomeric streptavidin. Technology, 1-7.
| None |