Difference between revisions of "Part:BBa K323214:Experience"
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
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===User Reviews=== | ===User Reviews=== | ||
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| + | [http://2012.igem.org/Team:Slovenia 2012 iGEM team Slovenia ] found out that [https://parts.igem.org/Part:BBa_K323214 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 [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782066 VP16] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782011 KRAB] domain. | ||
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| + | [[Image:Svn_12_NicTAL10vs12.PNG | 600 px]] | ||
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| + | '''Figure 1:'''Sequence alignment of NicTAL10 and NicTAL12, showing differences between constructs. | ||
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| + | [[Image:Svn_12_NicTAL_graf.png]] | ||
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| + | '''Figure 2:''' Results of testing [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782011 NicTAL:KRAB] repressor demonstrated better binding ability of NicTAL12. HEK293T cells were cotransfected with [https://parts.igem.org/Part:BBa_K782011 NicTAL12:KRAB] repressor under the control of a CMV promoter, and with a firefly luciferase reporter plasmid [https://parts.igem.org/Part:BBa_K782023 (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]. | ||
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<!-- Template for a user review | <!-- Template for a user review | ||
Latest revision as of 13:44, 26 September 2012
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K323214
User Reviews
[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].
UNIQc50459fef6b7c9ac-partinfo-00000000-QINU UNIQc50459fef6b7c9ac-partinfo-00000001-QINU