Part:BBa_K323214:Experience

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[http://2012.igem.org/Team:Slovenia 2012 iGEM team Slovenia ] found out that NicTAL part deposited in the Registry by the Slovenian iGEM2010 team was missing subdomain in DNA-binding domain, so our team added this missing part on N terminal sequence. Apart from adding N terminal sequence, we added HIS tag on N terminal end and NLS on terminal C end of NicTAL12 (Figure 1). This construct was later used for designing TAL-based activator and repressor by adding VP16 and KRAB domain.

Figure 1:Sequence alignment of NicTAL10 and NicTAL12, showing differences between constructs.

Results obtained by testing NicTAL12:KRAB repressor proved that NicTAL12 binds to its binding sites better than NicTAL10:KRAB.

Figure 2: Results of testing NicTAL12:KRAB repressor shows, that NicTAL12 binds to DNA better than NicTAL10.

UNIQ2d4deaa6fe296602-partinfo-00000000-QINU UNIQ2d4deaa6fe296602-partinfo-00000001-QINU