Part:BBa_K4907104

I0500-B0034-tmafp-B0015

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)

We assemble BBa_I0500, BBa_B0034, BBa_K4907004 and BBa_B0015 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
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1205
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  Illegal AgeI site found at 1438
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961