Difference between revisions of "Part:BBa K782032"

Revision as of 21:46, 26 September 2012

10x[TALA+TALB] operator_minimal promoter_fLuciferase

• TALA and TALB labels represents TAL effectors 1257 and 1297 from zebrafish experiments (Sander et al., 2011)
• DNA binding sites for individual TAL effectors are indicated with square brackets [ ].

Introduction

Transcription activation like (TAL) effectors are DNA binding proteins with a high specifity, built from tandem repeats, with near identical seqences, differing only in two amino acids in each repeat called “repeat variable diresidue” (RVD), which determine the specifity for a single nucleotide.(Scholze and Boch, 2011)

We designed our construct with 10 binding sites for TALA and TALB, upstream of a minimal promoter (like here). Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter.

Figure1:Schematic representation of our construct

Characterisation

10x[TALA+TALB] operator_minimal promoter was characterised with mCitrine reporter.

References

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

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

Sequence and Features