Part:BBa_K4765015
AnAFP
Contents
Introduction
Ammopiptanthus nanus antifreeze protein (AnAFP) is an antifreeze protein derived from Ammopiptanthus nanus. It protects the stability of membrane structures under cold conditions by binding to or dissociating from the membrane. It prevents the coagulation of macromolecules and plays a regulatory role by binding to or dissociating from calcium ions[1].
Usage and Biology
We heterologously expressed codon-optimized AnAFP in E.coli, endowing it with antifreeze capability.
Characterization
Sequencing map
|
| Figure1 Sequencing map of AnAFP
Sequencing was performed using the two primers：T7: 5-TAATACGACTCACTATAGG-3 and Kan-F: 5-ATTCTCACCGGATTCAGT-3. |
AnAFP Antifreeze Capability
|
| Figure2 Colonies at 0,24,48,72 and 96 hours of cold treatment at 0 °C
The experimental group consists of E. coli BL21 DE3 expressing AnAFP, the control group comprises E. coli BL21 DE3 transformed with the empty vector PET28. All the groups are cultured overnight. After reaching an OD value of 1.0, they are subjected to cold treatment at 0 °C.Samples are taken at 0, 24, 48, 72 and 96 hours during cold treatment and are diluted 10^5 times in ddH2O for CFU counting. CFU counting is performed with the help of imageJ. |
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 451
References
- ↑ Deng, L.-Q., Yu, H.-Q., Liu, Y.-P., Jiao, P.-P., Zhou, S.-F., Zhang, S.-Z., Li, W.-C., & Fu, F.-L. (2014). Heterologous expression of antifreeze protein gene AnAFP from Ammopiptanthus nanus enhances cold tolerance in Escherichia coli and tobacco. Gene, 539(1), 132-140.https://doi.org/10.1016/j.gene.2014.01.013
| protein |