Difference between revisions of "Part:BBa K1189023"
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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 | + | 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 and 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. |
| − | 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 | + | 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 <i>et al.</i>, 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 and Cathomen, 2012). |
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| − | TALE B was originally designed by Slovenia's 2012 iGEM team (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K782006"><b>BBa_K782006</b></a>). However, Slovenia's team made the part for use in eukaryotic cells. We modified the part so that it can be used in <i>E.coli</i> by removing the Kozak sequence and nuclear localization sequence, and codon optimizing it for <i>E.coli</i>. In addition, the sequence of TALE B reported by Slovenia was not completely accurate. We fixed the mutation in their sequence. </p> | + | TALE B was originally designed by Slovenia's 2012 iGEM team (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K782006"><b>BBa_K782006</b></a>). However, Slovenia's team made the part for use in eukaryotic cells. We modified the part so that it can be used in <i>E. coli</i> by removing the Kozak sequence and nuclear localization sequence, and codon optimizing it for <i>E. coli</i>. In addition, the sequence of TALE B reported by Slovenia was not completely accurate. We fixed the mutation in their sequence. </p> |
<p> This parts target sequence ([B]) is: tCTTCCGTTTCCACATCT </p> | <p> This parts target sequence ([B]) is: tCTTCCGTTTCCACATCT </p> | ||
Latest revision as of 01:03, 26 October 2013
TALE-B
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 and 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. 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 and Cathomen, 2012).
TALE B was originally designed by Slovenia's 2012 iGEM team (BBa_K782006). However, Slovenia's team made the part for use in eukaryotic cells. We modified the part so that it can be used in E. coli by removing the Kozak sequence and nuclear localization sequence, and codon optimizing it for E. coli. In addition, the sequence of TALE B reported by Slovenia was not completely accurate. We fixed the mutation in their sequence.
This parts target sequence ([B]) is: tCTTCCGTTTCCACATCT
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 394
Illegal XhoI site found at 1315 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]