Difference between revisions of "Part:BBa K782078"

Latest revision as of 22:35, 26 September 2012

10x[TALA+TALB] operator_CMV promoter_2xBFP:NLS

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

Introduction

Our construct contain TALA and TALB binding sites, which are adjacent to each other. Because of steric interference two different TAL regulators are unable to bind to its binding sites at once. TAL binding sites are cloned upstream of CMV promoter. Downstream of CMV is a double blue fluorescent protein with NLS domain, causing protein to migrate into the nucleus. BFP is a monomeric fluorescent protein with excitation maximum at 402 nm and emision maximum at 457 nm.

Figure 1: Schematic representation of the construct.

Characterization

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

BFP was obtained from Evrogen.

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.

Sequence and Features