Difference between revisions of "Part:BBa K2075020:Design"
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===Design Notes===
 
===Design Notes===
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This composite part includes an Intein and linker, the TAL-effector Ax7L-DS, a TEV Site and a Strep XT Tag. They are translated all together.  
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This part synthesize a transcription activator like (TAL) effector protein. They were found in Xanthomonas bacteria. These bacteria are plant pathogenes and infects plants via a secretion typ III system. In the plant cells these proteins binds promoter sites. This leads to a transcriptional activation of the genes behind the promoter. The proteins contains tandem repeats up to 34.5. Each repeat binds one nucleotide of the target sequenz. The 12th and 13th amino acid of each repeat leads to the specific binding of the DNA sequence.
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We circularised the TAL-effectors with the help of Inteins at the N- and C- Terminus of each TAL-effector.
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The 12th and 13th amino acid of each of the repeats from our TAL effector Ax7L-DS are: NI NN HD NI HD NG NI NG NI NG NI NI NI HD HD HD HD HD They bind the DNA sequence: A G/A C A C T A T A T A A A C C C C C
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The intein and linker are important to circularice the aminoacid sequence between the N- and C-terminal parts. Between the Intein sites is the TAL-effector and the TEV Site. There is also a strep XT tag.  
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When it is translated the intein sites react and form a circularised protein. All the parts between N- and C-Terminal Intein is part of the protein circle. In this circle the hole protein gains more stability.
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With the help of the strep XT tag the protein can be purified even if it is circularised. The Strep XT Tag is also usefull to detect the protein with an immunostain.
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The TEV Site give the opportunity to linearise the protein again wit the help oft he TEV protease. The linearised TALE can bind the specific DNA Sequence.  
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The 12th and 13th amino acid of each of the repeats from our TAL effector Ax7L-DS are: NI NN HD NI HD NG NI NG NI NG NI NI NI HD HD HD HD HD
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They bind the DNA sequence: A G/A C A C T A T A T A A A C C C C C
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===Source===
 
===Source===

Latest revision as of 18:59, 18 October 2016


TAL-effector Ax7L-DS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3202
    Illegal BamHI site found at 2608
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2547
    Illegal NgoMIV site found at 3142
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2622


Design Notes

This part synthesize a transcription activator like (TAL) effector protein. They were found in Xanthomonas bacteria. These bacteria are plant pathogenes and infects plants via a secretion typ III system. In the plant cells these proteins binds promoter sites. This leads to a transcriptional activation of the genes behind the promoter. The proteins contains tandem repeats up to 34.5. Each repeat binds one nucleotide of the target sequenz. The 12th and 13th amino acid of each repeat leads to the specific binding of the DNA sequence. We circularised the TAL-effectors with the help of Inteins at the N- and C- Terminus of each TAL-effector. The 12th and 13th amino acid of each of the repeats from our TAL effector Ax7L-DS are: NI NN HD NI HD NG NI NG NI NG NI NI NI HD HD HD HD HD They bind the DNA sequence: A G/A C A C T A T A T A A A C C C C C

Source

Source info goes here

References

References go here