Difference between revisions of "Part:BBa K4907004"

 
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<partinfo>BBa_K4907004 short</partinfo>
 
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===Biology===
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===Usage and Biology===
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SfIBP is a kind of antifreeze proteins (AFPs) found from <i>Shewanella frigidimarina</i>(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.
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===Usage and Design===
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To verify if LMT can translate SfIBP into the extracellular circumstance, this basic part was used to construct <partinfo>BBa_K4907103</partinfo> which was set as the negative control of <partinfo>BBa_K4907105</partinfo>.
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===Characterization===
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===Agarose gel electrophoresis (AGE)===
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<partinfo>BBa_I0500</partinfo>, <partinfo>BBa_B0034</partinfo>, <partinfo>BBa_K4907004</partinfo> and <partinfo>BBa_B0015</partinfo> were assembled to obtain the composite part <partinfo>BBa_K4907103</partinfo>, which are inserted 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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<center><html><img src="https://static.igem.wiki/teams/4907/wiki/parts/yyn/k4907004-fig1.png" width="400px"></html></center>
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<center>Fig. 1 Colony PCR of BBa_K4907103_pSB1C3 in <i>E. coli</i> BL21(DE3)</center>
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===Reference===
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#T. D. R. Vance, L. A. Graham, P. L. Davies, An ice-binding and tandem beta-sandwich domain-containing protein in <i>Shewanella frigidimarina</i> is a potential new type of ice adhesin. <i>The FEBS Journal</i> <b>285</b>, 1511-1527 (2018).
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Latest revision as of 05:03, 12 October 2023


sfibp


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 BBa_K4907103 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
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 634
    Illegal AgeI site found at 649
  • 1000
    COMPATIBLE WITH RFC[1000]