Difference between revisions of "Part:BBa K812200"
 
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This plasmid is a biobricked version of the plasmid pSC2+(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K812000">BBa_K812000</a>)with Elastase promoter. The CMV promoter was replaced by elastase promoter with SalI and HindIII restriction sites. This plasmid is made of a bacterial ampicilin resistant backbone. The Biobrick site is surrounded by a 5'UTR expression enhancer, and a poly-A tail with a SV40 3' UTR that enhance the stabilisation of the mRNA and the expression of the protein.</br>
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This plasmid is a biobricked version of the plasmid pCS2+(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K812000">BBa_K812000</a>)with Elastase promoter. The CMV promoter was replaced by elastase promoter with SalI and HindIII restriction sites. This plasmid is made of a bacterial ampicilin resistant backbone. The Biobrick site is surrounded by a 5'UTR expression enhancer, and a poly-A tail with a SV40 3' UTR that enhance the stabilisation of the mRNA and the expression of the protein.</br>
  
 
This plasmid is meant to be micro-injected in frog eggs or chicken eggs. This plasmid does not replicate, so it gets diluted and degraded as the animal grow, leaving no chance for DNA propagation to the breeding.</br>
 
This plasmid is meant to be micro-injected in frog eggs or chicken eggs. This plasmid does not replicate, so it gets diluted and degraded as the animal grow, leaving no chance for DNA propagation to the breeding.</br>
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To use this plasmid, you simply have to insert your protein between the EcoRI and PstI site. It is better to add a kozac sequence in front of your gene to enhance its expression. This plasmid contains the standard registry insert J04450 as a negative cloning control.</br></br>
 
To use this plasmid, you simply have to insert your protein between the EcoRI and PstI site. It is better to add a kozac sequence in front of your gene to enhance its expression. This plasmid contains the standard registry insert J04450 as a negative cloning control.</br></br>
  
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Elastase is a tissue specific promoter for the pancreas. This backbone is characterized using sfGFP (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K812031">BBa_K812031</a>) as reporter.</br>
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Elastase is a tissue specific promoter for the pancreas. This backbone is characterized using sfGFP (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K812031">BBa_K812031</a>) as reporter.</br>The result below is not strong enough and need more information to be sure that the plasmid works well (only one tadpole produced sfGFP into the pancreas).
  
 
Tadpole at stage 47 produced sfGFP in pancreas, you can compare the auto-fluorescence with the control and the picture with RFP filter compare to GFP.</br>
 
Tadpole at stage 47 produced sfGFP in pancreas, you can compare the auto-fluorescence with the control and the picture with RFP filter compare to GFP.</br>

Latest revision as of 08:52, 18 October 2012

Biobricked pCS2+ plasmid with Elastase promoter for frogs and chicken

This plasmid is a biobricked version of the plasmid pCS2+(BBa_K812000)with Elastase promoter. The CMV promoter was replaced by elastase promoter with SalI and HindIII restriction sites. This plasmid is made of a bacterial ampicilin resistant backbone. The Biobrick site is surrounded by a 5'UTR expression enhancer, and a poly-A tail with a SV40 3' UTR that enhance the stabilisation of the mRNA and the expression of the protein.
This plasmid is meant to be micro-injected in frog eggs or chicken eggs. This plasmid does not replicate, so it gets diluted and degraded as the animal grow, leaving no chance for DNA propagation to the breeding.
To use this plasmid, you simply have to insert your protein between the EcoRI and PstI site. It is better to add a kozac sequence in front of your gene to enhance its expression. This plasmid contains the standard registry insert J04450 as a negative cloning control.

Elastase is a tissue specific promoter for the pancreas. This backbone is characterized using sfGFP (BBa_K812031) as reporter.
The result below is not strong enough and need more information to be sure that the plasmid works well (only one tadpole produced sfGFP into the pancreas). Tadpole at stage 47 produced sfGFP in pancreas, you can compare the auto-fluorescence with the control and the picture with RFP filter compare to GFP.
perdu


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3291
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 248
    Illegal NotI site found at 3297
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3291
    Illegal BamHI site found at 3276
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3291
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3291
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3306
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 2611
    Illegal NgoMIV site found at 3032
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 1634
    Illegal SapI site found at 551