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.

Figure 2: Results of testing NicTAL:KRAB repressor demonstrated better binding ability of NicTAL12. HEK293T cells were cotransfected with NicTAL12:KRAB repressor under the control of a CMV promoter, and with a firefly luciferase reporter plasmid (BBa_K782023) containing 12 DNA-binding sites for NicTAL repressor upstream the CMV promoter. Check detailed results on [http://2012.igem.org/Team:Slovenia/TheSwitchDesignedTALregulators iGEM 2012 team Slovenia wiki].

UNIQ0b5dcea1c41955eb-partinfo-00000000-QINU UNIQ0b5dcea1c41955eb-partinfo-00000001-QINU