Difference between revisions of "Part:BBa K782068"
 
(8 intermediate revisions by 3 users not shown)
Line 3: Line 3:
  
 
* TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
 
* TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
 +
* DNA binding sites for individual TAL effectors are indicated with square brackets [ ].
 +
  
 
==Introduction==
 
==Introduction==
  
−
Transcription activation like (TAL) effectors are bacterial plant pathogen transcription factors that bind to DNA by recognizing a specific DNA sequence in which each base pair binds a single tandem repeat in in the TAL DNA-binding domain. A tandem TAL repeat contains 33 to 35 amino acids, where the 12th and the 13th amino acid, called a “repeat variable diresidue” (RVD)  are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011).  All TAL repeats have almost identical sequences, differing only in the RVDs. This modularity of TAL effector binding domains therefore makes them a perfect tool to target specific DNA sequences by designing specific binding domains for a selected TAL effector. We designed four consecutive specific binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782011 NicTAL12:KRAB] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782009 TALD:KRAB] (Figure 1).  
+
Our construct contains four specific binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782007 NicTAL12] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782005 TALD].  
−
 
+
−
Single binding sequence for NicTAL is: TCTATCAATGATAGA
+
Single binding sequence for NicTAL12 is: TCTATCAATGATAGA  
  
 
Single binding sequence for TALD is: TCGTCCAATAGCTTCTC
 
Single binding sequence for TALD is: TCGTCCAATAGCTTCTC
  
  
−
[[Image:4xNic+4xD.png]]
 
  
−
'''Figure 1.''' Shematic representation of four consecutive specific binding sites for NicTAL and TALD.
+
[[Image:4xNic+4xD.png‎]]
  
 +
'''Figure 1.''' Shematic representation of four consecutive specific binding sites for NicTAL12 and TALD.
  
  
 
* Binding sites for TAL effectors were ordered from IDT.  
 
* Binding sites for TAL effectors were ordered from IDT.  
 +
 +
 +
==Characterization==
 +
 +
Specific TAL binding sites were further characterized with different reporter constructs.
 +
 +
'''Results:'''
 +
 +
* [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782025 BBa_K782025]
  
  
Line 26: Line 37:
 
Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.
 
Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.
  
−
Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698  
+
Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698.
  
  

Latest revision as of 22:51, 26 September 2012

4x[NicTAL]+4x[TALD] operator

  • TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
  • DNA binding sites for individual TAL effectors are indicated with square brackets [ ].


Introduction

Our construct contains four specific binding sites for NicTAL12 and TALD.

Single binding sequence for NicTAL12 is: TCTATCAATGATAGA

Single binding sequence for TALD is: TCGTCCAATAGCTTCTC


4xNic+4xD.png

Figure 1. Shematic representation of four consecutive specific binding sites for NicTAL12 and TALD.


  • Binding sites for TAL effectors were ordered from IDT.


Characterization

Specific TAL binding sites were further characterized with different reporter constructs.

Results:


References

Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.

Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 35
    Illegal AgeI site found at 235
  • 1000
    COMPATIBLE WITH RFC[1000]