Difference between revisions of "Part:BBa K2332313"
 
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__NOTOC__
 
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<partinfo>BBa_K2332313 short</partinfo>
 
<partinfo>BBa_K2332313 short</partinfo>
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This composite part is a device which efficiently encodes E-cadherin, a calcium-dependent cell adhesion molecule that functions in the establishment and maintenance of epithelial cell morphology during embryongenesis and adulthood. The encoded preproprotein undergoes proteolytic processing to generate a mature protein.
 
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
 
{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
 
! colspan="2" style="background:#FFBF00;"|E-cadherin (Preproprotein) with CMV promoter & bGH poly(A) tail
 
! colspan="2" style="background:#FFBF00;"|E-cadherin (Preproprotein) with CMV promoter & bGH poly(A) tail
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|'''Use in'''
 
|'''Use in'''
 
|Mammalian cells
 
|Mammalian cells
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|-
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|'''Chassis Tested'''
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|Chinese Hamster Ovary (CHO)
 
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|'''Abstraction Hierarchy'''
 
|'''Abstraction Hierarchy'''
 
|Device
 
|Device
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|-
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|'''Related Parts'''
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|[https://parts.igem.org/wiki/index.php?title=Part:BBa_I712004 BBa_I712004], [https://parts.igem.org/Part:BBa_K2332312 BBa_K2332312], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678019 BBa_K678019]
 
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|'''RFC standard'''
 
|'''RFC standard'''
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|'''Submitted by'''
 
|'''Submitted by'''
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|[http://2017.igem.org/Team:UCL]
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|[http://2017.igem.org/Team:UCL UCL iGEM 2017]
 
|}
 
|}
  
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This composite part is a device which efficiently encodes E-cadherin, a calcium-dependent cell adhesion molecule that functions in the establishment and maintenance of epithelial cell morphology during embryongenesis and adulthood. The encoded preproprotein undergoes proteolytic processing to generate a mature protein.
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[[File:CMV Promoter - 5'-UTR - E-cadherin.png|700px|thumb|left|'''Figure 1: Construct Design.''' The CMV promoter was fused with the E-cadherin coding part by creating a 5'-UTR (untranslated region) which is essential for efficient translation. The resulting fusion construct contains a T7 promoter according to pcDNA3 standard, a standard mammalian expression plasmid. The CMV promoter and bGH terminator were chosen to ensure efficient gene expression. ]]
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===Experimental approach===
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For testing this device we used CHO cells as our chassis of choice, which naturally lack E-cadherin expression but still maintain alpha- and beta-catenin expression. By chosing CHO cells as our chassis we, therefore, eliminated background noise.
  
  
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
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Latest revision as of 14:42, 8 October 2017


E-cadherin (Preproprotein) with CMV promoter & bGH poly(A) tail

E-cadherin (Preproprotein) with CMV promoter & bGH poly(A) tail
Function Cell-Cell Adhesion
Use in Mammalian cells
Chassis Tested Chinese Hamster Ovary (CHO)
Abstraction Hierarchy Device
Related Parts BBa_I712004, BBa_K2332312, BBa_K678019
RFC standard RFC10 & RFC23 compatible
Backbone pSB1C3
Submitted by [http://2017.igem.org/Team:UCL UCL iGEM 2017]


This composite part is a device which efficiently encodes E-cadherin, a calcium-dependent cell adhesion molecule that functions in the establishment and maintenance of epithelial cell morphology during embryongenesis and adulthood. The encoded preproprotein undergoes proteolytic processing to generate a mature protein.



Figure 1: Construct Design. The CMV promoter was fused with the E-cadherin coding part by creating a 5'-UTR (untranslated region) which is essential for efficient translation. The resulting fusion construct contains a T7 promoter according to pcDNA3 standard, a standard mammalian expression plasmid. The CMV promoter and bGH terminator were chosen to ensure efficient gene expression.












Experimental approach

For testing this device we used CHO cells as our chassis of choice, which naturally lack E-cadherin expression but still maintain alpha- and beta-catenin expression. By chosing CHO cells as our chassis we, therefore, eliminated background noise.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3212
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1414
    Illegal BamHI site found at 1490
    Illegal BamHI site found at 1606
    Illegal BamHI site found at 2530
    Illegal BamHI site found at 2832
    Illegal XhoI site found at 2214
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 870
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 727
    Illegal BsaI site found at 1127
    Illegal BsaI site found at 1534
    Illegal BsaI site found at 2076
    Illegal BsaI.rc site found at 968