Difference between revisions of "Part:BBa K929302"

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AID is known to be responsible for somatic hypermutation and the class-switch recombination of immunoglobulin in B cells. This enzyme of 28 kDa originally occurs in B cells but does also show activity after transfection into CHO cells. AID induces the deamination of cytidine to uridine at actively transcribed single strand DNA. The replacement of cytidine by uridine leads to a mismatch during DNA replication and integrates a single base substitution predominantly in the immunoglobulin genes.
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AID is known to be responsible for somatic hypermutation and the class-switch recombination of immunoglobulin in B cells. This enzyme of 28 kDa originally occurs in B cells but does also show activity after transfection into CHO cells. AID induces the deamination of cytidine to uridine at actively transcribed single strand DNA. The replacement of cytidine by uridine leads to a mismatch during DNA replication and integrates a single base substitution predominantly in the immunoglobulin genes.<br>
[[image:UP_12_Potstdam_Standard.png|400px|center|thumb|Relative ligation success of the four conditions]]
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To generate this part, we used the Potsdam Standard for cloning the AID into the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K929301 Potsdam Standard Backbone]. Here, we tested four different ligation conditions: 1. with electrophoretic unpurified, digested Potsdam Standard backbone with and without ligase and 2. with electrophoretic purified digested Potsdam Standard backbone with and without ligase. The results are summarized in fig. 1. 
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[[image:UP_12_Potstdam_Standard.png|400px|center|thumb|Fig. 1: Relative ligation success of the four conditions]]
 
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Revision as of 19:57, 26 September 2012

AID in Potsdam Standard



AID is known to be responsible for somatic hypermutation and the class-switch recombination of immunoglobulin in B cells. This enzyme of 28 kDa originally occurs in B cells but does also show activity after transfection into CHO cells. AID induces the deamination of cytidine to uridine at actively transcribed single strand DNA. The replacement of cytidine by uridine leads to a mismatch during DNA replication and integrates a single base substitution predominantly in the immunoglobulin genes.
To generate this part, we used the Potsdam Standard for cloning the AID into the Potsdam Standard Backbone. Here, we tested four different ligation conditions: 1. with electrophoretic unpurified, digested Potsdam Standard backbone with and without ligase and 2. with electrophoretic purified digested Potsdam Standard backbone with and without ligase. The results are summarized in fig. 1.

Fig. 1: Relative ligation success of the four conditions










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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 84
    Illegal SapI site found at 185