Difference between revisions of "Part:BBa K1189022:Design"
 
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===Design Notes===
 
===Design Notes===
  
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Transcriptor Activator-Like Effectors (TALEs) are proteins produced by bacteria of the genus Xanthomonas and secreted into plant cells. These naturally occurring TALEs play a key role in bacterial infection, as they are responsible for up regulation of the host genes required for pathogenic growth and expansion (Mussolino & Cathomen, 2012). These special bacterial plant pathogen proteins bind to DNA by specifically recognizing one base pair with a single tandem repeat in their DNA-binding domain. TALEs are an advantageous tool in synthetic biology because they can be modified to bind to a chosen DNA sequence.
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On the iGEM parts registry we found TALA (BBa_K782004) that the Slovenian team synthesized for a previous project. TALEs are very large proteins and take a long time to design and synthesize. Therefore, we decided to use these TALEs to test our system. We also saw this as an opportunity to use and build upon parts made by former iGEM teams. The TALE also had a kozak sequence at the front of the sequence, we used PCR to remove the kozak sequence at the front of the TALE, so it would not inhibit the expression
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The central region of the protein, also termed repeat region, mediates DNA recognition through tandem repeats of 33 to 35 amino acids residues each (Bogdanove et al., 2010). The binding domain usually comprises 15.5 to 19.5 single repeats. The last repeat, close to the C-terminus, is called “half-repeat” because it is only approximately 20 amino acids in length. Although the modules have conserved sequences, polymorphisms are found in residues 12 and 13, the “repeat-variable di-residue” (RVD). RVDs are specific for a single nucleotide; therefore, 19.5 repeat units target a specific 20-nucleotide sequence in the DNA (Mussolino & Cathomen, 2012).
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On the iGEM parts registry we found TALA (BBa_K782004) that the Slovenian team synthesized for a previous project. TALEs are very large proteins and take a long time to design and synthesize. Therefore, we decided to use these TALEs to test our system. We also saw this as an opportunity to use and build upon parts made by former iGEM teams.
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===Source===
 
===Source===
  
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===References===
 
===References===
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<li>Bogdanove, A. J., Schornack, S., & Lahaye, T. (2010). TAL effectors: finding plant genes for disease and defense. <i>Current Opinion in Plant Biology, 13</i>(4), 394–401. doi:10.1016/j.pbi.2010.04.010
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<li>Mussolino, C., & Cathomen, T. (2012). TALE nucleases: tailored genome engineering made easy. <i>Current Opinion in Biotechnology, 23</i>(5), 644–50. doi:10.1016/j.copbio.2012.01.013

Latest revision as of 00:55, 26 October 2013

TALE-A


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1696
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2425
    Illegal AgeI site found at 451
    Illegal AgeI site found at 1032
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

On the iGEM parts registry we found TALA (BBa_K782004) that the Slovenian team synthesized for a previous project. TALEs are very large proteins and take a long time to design and synthesize. Therefore, we decided to use these TALEs to test our system. We also saw this as an opportunity to use and build upon parts made by former iGEM teams. The TALE also had a kozak sequence at the front of the sequence, we used PCR to remove the kozak sequence at the front of the TALE, so it would not inhibit the expression

Source

Parts Registry BBa_K782004

References

  • Bogdanove, A. J., Schornack, S., & Lahaye, T. (2010). TAL effectors: finding plant genes for disease and defense. Current Opinion in Plant Biology, 13(4), 394–401. doi:10.1016/j.pbi.2010.04.010
  • Mussolino, C., & Cathomen, T. (2012). TALE nucleases: tailored genome engineering made easy. Current Opinion in Biotechnology, 23(5), 644–50. doi:10.1016/j.copbio.2012.01.013