Difference between revisions of "Part:BBa K323214:Experience"

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===User Reviews===
 
===User Reviews===
  
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[http://2012.igem.org/Team:Slovenia 2012 iGEM team Slovenia ] discovered 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).  
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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.
  
 
[[Image:Svn_12_NicTAL10vs12.PNG | 600 px]]
 
[[Image:Svn_12_NicTAL10vs12.PNG | 600 px]]
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Results obtained by testing [https://parts.igem.org/Part:BBa_K782011#Characterization NicTAL12:KRAB] repressor proved that NicTAL12 binds to its binding sites better than NicTAL10.  
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Results obtained by testing [https://parts.igem.org/Part:BBa_K782011#Characterization NicTAL12:KRAB] repressor proved that NicTAL12 binds to its binding sites better than NicTAL10:KRAB.  
  
 
[[Image:Svn_12_NicTAL_graf.png]]
 
[[Image:Svn_12_NicTAL_graf.png]]
  
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'''Figure 2:''' Results of testing NicTAL12:KRAB repressor shows, that NicTAL12 binds to DNA better than NicTAL10.
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'''Figure 2:''' Results of testing [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782011 NicTAL12:KRAB] repressor shows, that NicTAL12 binds to DNA better than NicTAL10.
  
  

Revision as of 17:40, 25 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.


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

Svn 12 NicTAL graf.png

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



UNIQcdcf27addd3ba7c9-partinfo-00000000-QINU UNIQcdcf27addd3ba7c9-partinfo-00000001-QINU