Difference between revisions of "Part:BBa K782067"
Line 6: Line 6:
 
==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 two consecutive specific binding sites for NicTAL12 and TALD (Figure 1).
+
Our construct contains two 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 NicTAL12 is: TCTATCAATGATAGA
  
−
Single binding sequence for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782007 NicTAL12] is: TCTATCAATGATAGA 
+
Single binding sequence for TALD is: TCGTCCAATAGCTTCTC
  
−
Single binding sequence for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782005 TALD] is: TCGTCCAATAGCTTCTC
 
  
  
Line 18: Line 19:
  
  
 +
==Characterization==
 +
 +
 +
Results: Specific TAL binding sites were further characterized with different reporter constructs.
  
 
* Binding sites for TAL effectors were ordered from IDT.  
 
* Binding sites for TAL effectors were ordered from IDT.  

Revision as of 07:51, 26 September 2012

2x[NicTAL]+2x[TALD] operator

  • TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).

Introduction

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

Single binding sequence for NicTAL12 is: TCTATCAATGATAGA

Single binding sequence for TALD is: TCGTCCAATAGCTTCTC


2xNic+2xD.png

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


Characterization

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

  • Binding sites for TAL effectors were ordered from IDT.


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 105
  • 1000
    COMPATIBLE WITH RFC[1000]