Difference between revisions of "Part:BBa K4390059"

 
 
(3 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K4390059 short</partinfo>
 
<partinfo>BBa_K4390059 short</partinfo>
  
−
-
+
'''This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard [[Help:Standards/Assembly/Type_IIS|which is also accepted by iGEM.]]'''
  
−
<!-- Add more about the biology of this part here
+
'''This is a level 1 part formed by assembly of the following level 0 parts:'''
−
===Usage and Biology===
+
{| class="wikitable" style="margin:auto"
 +
|-
 +
| Promoter || [[part:BBa_J23100|J23100]]
 +
|-
 +
| RBS || [[part:BBa_B0034|B0034]]
 +
|-
 +
| N part || [[part:BBa_K3946001|K3946001]]
 +
|-
 +
| O part || [[part:BBa_K4390007|K4390007]]
 +
|-
 +
| C part || [[part:BBa_I746916|I746916]]
 +
|-
 +
| Terminator || [[part:BBa_K4390001|K4390001]]
 +
|}
 +
 
 +
==Usage and Biology==
 +
The green fluorescent protein (GFP) is a protein that can produce bright green colour. Due to these properties, it has always been used as reporter of protein expression. Protein fused with sfGFP will turn the colony green if expressed, which can be used to identify the ability of certain genes to be expressed in other organism like E. coli. Superfolder GFP (sfGFP) is a more robustly folded version of GFP, which is derived from GFP that often misfolds when expressed as fusions with other proteins. Superfolder GPF shows improved tolerance of circular permutation, greater resistance to chemical denaturants and improved folding kinetics.
 +
 
 +
This is one of the three types of silica-binding tags we explored for immobilising metallothioneins to silica beads. Car9 was also a silica-tag part used by the 2021 Edinburgh OG Team.
 +
 
 +
The green-fluorescent recombinant protein was used as an indicator to show qualitatively and quantitatively whether or not the Car9 peptide has affinity towards silica beads. This was assayed by comparing the green fluorescence intensity between the initial lysate and the supernatant after addition of the silica beads. There should be a significant reduction in fluorescence intensity to prove that the Car9 peptide indeed binds to silica beads and thus, immobilises the sfGFP fused to it.
  
 
<!-- -->
 
<!-- -->
−
<span class='h3bb'>Sequence and Features</span>
+
 
 +
==<span class='h3bb'>Sequence and Features</span>==
 
<partinfo>BBa_K4390059 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4390059 SequenceAndFeatures</partinfo>
  

Latest revision as of 23:41, 13 October 2022


Car9-tagged sfGFP

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

This is a level 1 part formed by assembly of the following level 0 parts:

Promoter J23100
RBS B0034
N part K3946001
O part K4390007
C part I746916
Terminator K4390001

Usage and Biology

The green fluorescent protein (GFP) is a protein that can produce bright green colour. Due to these properties, it has always been used as reporter of protein expression. Protein fused with sfGFP will turn the colony green if expressed, which can be used to identify the ability of certain genes to be expressed in other organism like E. coli. Superfolder GFP (sfGFP) is a more robustly folded version of GFP, which is derived from GFP that often misfolds when expressed as fusions with other proteins. Superfolder GPF shows improved tolerance of circular permutation, greater resistance to chemical denaturants and improved folding kinetics.

This is one of the three types of silica-binding tags we explored for immobilising metallothioneins to silica beads. Car9 was also a silica-tag part used by the 2021 Edinburgh OG Team.

The green-fluorescent recombinant protein was used as an indicator to show qualitatively and quantitatively whether or not the Car9 peptide has affinity towards silica beads. This was assayed by comparing the green fluorescence intensity between the initial lysate and the supernatant after addition of the silica beads. There should be a significant reduction in fluorescence intensity to prove that the Car9 peptide indeed binds to silica beads and thus, immobilises the sfGFP fused to it.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 145