Difference between revisions of "Part:BBa K747102"
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<partinfo>BBa_K747102 short</partinfo>
 
<partinfo>BBa_K747102 short</partinfo>
  
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This part can be used to perform to creates site specific mutations in DNA by inserting a twelfe nucleotide binding Tal-Effector. See Bba_K747001 to Bba_K747095. The AID removes the amino group from a cytidine base.
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This Part was build in collaboration with Team Potsdam, which is working on a CHO cell-based project for generating antibodies. Hypermutation should be accomplished by cotransfection of the activation-induced cytidine deaminase (AID). Since it is obviously preferable that the activity of the AID is focussed on the antibody gene, we decided to put our expertise in our two distinct research fields together and produce a TAL-AID fusion protein that binds to the antibody gene-containing plasmid. Using the Plug-and-Play Effector Cassette (PPEC) in our [https://parts.igem.org/wiki/index.php?title=Part:BBa_K747099 Eukaryotic TAL expression vector], it was quite easy to fuse the AID gene to our TAL scaffold. The next step, namely targeting the TAL-AID construct to the antibody plasmid, was a child's play with our GATE kit. So we produced a TAL-AID that specifically binds to the sequence T G C G G T C T A T T A C T and currently test it in collaboration with the Potsdam Bioware Team. We hope that we will be able to show you the new data in Boston.  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:53, 31 October 2012

pTAL-AID

This Part was build in collaboration with Team Potsdam, which is working on a CHO cell-based project for generating antibodies. Hypermutation should be accomplished by cotransfection of the activation-induced cytidine deaminase (AID). Since it is obviously preferable that the activity of the AID is focussed on the antibody gene, we decided to put our expertise in our two distinct research fields together and produce a TAL-AID fusion protein that binds to the antibody gene-containing plasmid. Using the Plug-and-Play Effector Cassette (PPEC) in our Eukaryotic TAL expression vector, it was quite easy to fuse the AID gene to our TAL scaffold. The next step, namely targeting the TAL-AID construct to the antibody plasmid, was a child's play with our GATE kit. So we produced a TAL-AID that specifically binds to the sequence T G C G G T C T A T T A C T and currently test it in collaboration with the Potsdam Bioware Team. We hope that we will be able to show you the new data in Boston.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal suffix found in sequence at 3200
    Illegal XbaI site found at 3253
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1999
    Illegal SpeI site found at 3201
    Illegal PstI site found at 3215
    Illegal NotI site found at 3208
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3241
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal suffix found in sequence at 3201
    Illegal XbaI site found at 3253
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3253
    Illegal SpeI site found at 3201
    Illegal PstI site found at 3215
    Illegal NgoMIV site found at 3823
    Illegal NgoMIV site found at 5106
    Illegal NgoMIV site found at 5389
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2071
    Illegal BsaI.rc site found at 1417
    Illegal BsaI.rc site found at 2233
    Illegal BsaI.rc site found at 2722
    Illegal BsaI.rc site found at 5601
    Illegal SapI site found at 2823
    Illegal SapI.rc site found at 4955
    Illegal SapI.rc site found at 5165