Difference between revisions of "Part:BBa K3782000"
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=Usage and Biology=
 
=Usage and Biology=
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The FfIBP protein coding region was used in the following composite parts (add links). It was expressed in <i>E. coli</i> strain BL21 (DE3) and purified. Various tests and assays were performed to characterize and verify the functionality of this ice-binding protein. FfIBP has moderate TH and IRI activity, and it can bind to ice crystals and inhibit their growth. The aim of our project was to use it on its own or in a mixture of other antifreeze proteins to develop a solution which could be applied on sensitive plant tissues and thereby protect crops from frost damage.
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FfIBP is naturally produced by the Gram-negative Antarctic bacterium <i>Flavobacterium frigoris</i> PS1. Its TH activity is around 2.5 K at 50 µM<ref>Do H, Lee JH, Lee SG, Kim HJ. Crystallization and preliminary X-ray crystallographic analysis of an ice-binding protein (FfIBP) from Flavobacterium frigoris PS1. Acta Crystallogr Sect F Struct Biol Cryst Commun [Internet]. 2012 Jul [cited 2021 Jul 7];68(Pt 7):806. Available from: /pmc/articles/PMC3388927/</ref>.
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'''!!!ADD Biological context and overall mechanism of the part!!!'''
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=Characterization=
 
=Characterization=
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<h3>Cloning</h3>
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<h3>Expression and Purification</h3>
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<h3>ISF Assay</h3>
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<h3>FDT Assay</h3>
 
=Sequence and Features=
 
=Sequence and Features=
 
<partinfo>BBa_K3782000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3782000 SequenceAndFeatures</partinfo>
 
=References=
 
=References=
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{{reflist}}
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FfIBP is naturally produced by the Gram-negative Antarctic bacterium <i>Flavobacterium frigoris</i> PS1. Its TH activity is around 2.5 K at 50 µM.
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Revision as of 13:57, 9 October 2021

Flavobacterium frigoris strain PS1 ice-binding protein gene

FfIBP

FfIBP is a protein coding region that codes for an ice binding protein (IBP). IBPs, or more specifically antifreeze proteins (AFP), can bind to ice crystals and thereby prevent further ice growth. They are produced by organisms to survive in extremely cold environments. Activities of AFPs can be characterized by their thermal hysteresis (TH) or by their ice recrystallization inhibition (IRI). TH activity corresponds to the lowering of the freezing point without changing the melting point of a solution. IRI activity inhibits the growth of large ice crystals at the expense of smaller ones. The combination of these activities, which vary depending on the protein structure, prevents the freezing of body fluids and cell damage in organisms that live in environments with extremely cold temperatures.

Profile

Name FfIBP
Base pairs 744
Number of amino acids 247
Molecular weight 25.35 kDa
Origin Flavobacterium frigoris strain PS1, synthetic

Usage and Biology

The FfIBP protein coding region was used in the following composite parts (add links). It was expressed in E. coli strain BL21 (DE3) and purified. Various tests and assays were performed to characterize and verify the functionality of this ice-binding protein. FfIBP has moderate TH and IRI activity, and it can bind to ice crystals and inhibit their growth. The aim of our project was to use it on its own or in a mixture of other antifreeze proteins to develop a solution which could be applied on sensitive plant tissues and thereby protect crops from frost damage.

FfIBP is naturally produced by the Gram-negative Antarctic bacterium Flavobacterium frigoris PS1. Its TH activity is around 2.5 K at 50 µM[1].

!!!ADD Biological context and overall mechanism of the part!!!

Characterization

Cloning

Expression and Purification

ISF Assay

FDT Assay

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 269
    Illegal PstI site found at 350
    Illegal PstI site found at 530
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 269
    Illegal PstI site found at 350
    Illegal PstI site found at 530
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 269
    Illegal PstI site found at 350
    Illegal PstI site found at 530
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 269
    Illegal PstI site found at 350
    Illegal PstI site found at 530
  • 1000
    COMPATIBLE WITH RFC[1000]

References

Template:Reflist
  1. ↑ Do H, Lee JH, Lee SG, Kim HJ. Crystallization and preliminary X-ray crystallographic analysis of an ice-binding protein (FfIBP) from Flavobacterium frigoris PS1. Acta Crystallogr Sect F Struct Biol Cryst Commun [Internet]. 2012 Jul [cited 2021 Jul 7];68(Pt 7):806. Available from: /pmc/articles/PMC3388927/