Part:BBa_K4907103

I0500-B0034-sfibp-B0015

Biology

SfIBP is a kind of antifreeze proteins (AFPs) found from Shewanella frigidimarina(1). It is capable of preventing liquid from freezing within a certain temperature range and inhibiting ice growth, which is defined as Thermal Hysteresis (TH) and ice recrystallization inhibition (IRI) respectively. Additionally, the IRI activity of SfIBP is much higher than other AFPs.

Usage and Design

To verify if LMT can translate SfIBP into the extracellular circumstance, this basic part was used to construct <parinfo>BBa_K4907103</partinfo> which was set as the negative control of BBa_K4907105.

Characterization

Agarose gel electrophoresis (AGE)

BBa_I0500, BBa_B0034, BBa_K4907004 and BBa_B0015 were assembled to obtain the composite part BBa_K4907103, which are inserted 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_K4907103_pSB1C3 in E. coli BL21(DE3)

Reference

1. T. D. R. Vance, L. A. Graham, P. L. Davies, An ice-binding and tandem beta-sandwich domain-containing protein in Shewanella frigidimarina is a potential new type of ice adhesin. The FEBS Journal 285, 1511-1527 (2018).

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 1870
  Illegal AgeI site found at 1885
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961