Difference between revisions of "Part:BBa K782026"
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<partinfo>BBa_K782026 short</partinfo> | <partinfo>BBa_K782026 short</partinfo> | ||
− | + | * TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011). | |
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+ | ==Introduction== | ||
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+ | Transcription activation like (TAL) effectors are proteins able to specifically bind desired DNA sequence. The central domain of the protein is constructed from variable number of tandem repeats differing only in two amino acids. The 12th and the 13th amino acid are called a “repeat variable diresidue” (RVD) and are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011). This modularity of TAL effector binding domains therefore makes them a perfect tool to target specific DNA sequences. | ||
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+ | Our construct contains seven specific binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782004 TALA] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782006 TALB] upstream of minimal promoter. Downstream of minimal promoter we cloned yellow fluorescent protein mCitrine an easy detectable monomer with excitation maximum at 516 nm and emission maximum at 529 nm (Figure 1). | ||
+ | After binding of [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782065 TALA:VP16] or [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782012 TALB:VP16] on binding sites, an activation of reporter protein mCitrine occurs. | ||
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Revision as of 15:19, 26 September 2012
7x[TALA+TALB] operator_minimal promoter_mCitrine
- TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
Introduction
Transcription activation like (TAL) effectors are proteins able to specifically bind desired DNA sequence. The central domain of the protein is constructed from variable number of tandem repeats differing only in two amino acids. The 12th and the 13th amino acid are called a “repeat variable diresidue” (RVD) and are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011). This modularity of TAL effector binding domains therefore makes them a perfect tool to target specific DNA sequences.
Our construct contains seven specific binding sites for TALA and TALB upstream of minimal promoter. Downstream of minimal promoter we cloned yellow fluorescent protein mCitrine an easy detectable monomer with excitation maximum at 516 nm and emission maximum at 529 nm (Figure 1). After binding of TALA:VP16 or TALB:VP16 on binding sites, an activation of reporter protein mCitrine occurs.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 488
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 192
Illegal AgeI site found at 55
Illegal AgeI site found at 395 - 1000COMPATIBLE WITH RFC[1000]