Difference between revisions of "Part:BBa K323214:Experience"
 
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[[Image:Svn_12_NicTAL_graf.png]]
 
[[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.  
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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].
  
  

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.

Svn 12 NicTAL10vs12.PNG

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


Svn 12 NicTAL graf.png

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].



UNIQee74a01f33419399-partinfo-00000000-QINU UNIQee74a01f33419399-partinfo-00000001-QINU