Difference between revisions of "Part:BBa K4907005"
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===Characterization===
 
===Characterization===
 
====Agarose gel electrophoresis (AGE)====
 
====Agarose gel electrophoresis (AGE)====
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We assemble <partinfo>BBa_I0500</partinfo>, <partinfo>BBa_B0034</partinfo>, <partinfo>BBa_K4907004</partinfo> and <partinfo>BBa_B0015</partinfo> to obtain the composite part <partinfo>BBa_K4907104</partinfo>, which are assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmids are transformed into <i>E. coli</i> BL21(DE3), then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.
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<partinfo>BBa_I0500</partinfo>, <partinfo>BBa_B0034</partinfo>, <partinfo>BBa_K4907004</partinfo> and <partinfo>BBa_B0015</partinfo> was assembled to obtain the composite part <partinfo>BBa_K4907104</partinfo>, which are assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmids are transformed into <i>E. coli</i> BL21(DE3), then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.
 
<center><html><img src="https://static.igem.wiki/teams/4907/wiki/parts/yyn/k4907005-fig1.png" width="400px"></html></center>
 
<center><html><img src="https://static.igem.wiki/teams/4907/wiki/parts/yyn/k4907005-fig1.png" width="400px"></html></center>
 
<center>Fig. 1 Colony PCR of BBa_K4907104_pSB1C3 in <i>E. coli</i> BL2(DE3)</center>
 
<center>Fig. 1 Colony PCR of BBa_K4907104_pSB1C3 in <i>E. coli</i> BL2(DE3)</center>

Revision as of 13:55, 10 October 2023


tmafp

Part: BBa_K4907005 tmafp

Biology

TmAFP, found in Tenebrio molitor, is one of the antifreeze proteins (AFPs) (1). It is capable of preventing liquid freezing and inhibiting ice growth, which is defined as Thermal Hysteresis (TH) and ice recrystallization inhibition (IRI) respectively. Notably, TmAFP possesses the highest TH activity among all AFPs that have been found.

Usage

To verify if LMT can translate TmAFP into the extracellular circumstance, this basic part was used to construct <parinfo>BBa_K4907104</partinfo> which was set as the negative control of BBa_K4907106.

Characterization

Agarose gel electrophoresis (AGE)

BBa_I0500, BBa_B0034, BBa_K4907004 and BBa_B0015 was assembled to obtain the composite part BBa_K4907104, which are assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmids are transformed into E. coli BL21(DE3), then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.

Fig. 1 Colony PCR of BBa_K4907104_pSB1C3 in E. coli BL2(DE3)

Reference

  1. U. S. Midya, S. Bandyopadhyay, Elucidating the Sluggish Water Dynamics at the Ice-Binding Surface of the Hyperactive Tenebrio molitor Antifreeze Protein. J. Phys. Chem. B 127, 121-132 (2023).


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 202
  • 1000
    COMPATIBLE WITH RFC[1000]