Difference between revisions of "Part:BBa K782086"
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* TALA and TALB labels represents TAL effector 1257 and 1297 from zebrafish experiments (Sander et al., 2011). | * 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 [ ]. | ||
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==Introduction== | ==Introduction== | ||
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'''Figure 1:''' Schematic representation of the construct. | '''Figure 1:''' Schematic representation of the construct. | ||
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| + | * Binding sites for TAL effectors were ordered from IDT. | ||
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==References== | ==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 | + | 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 23:02, 26 September 2012
10x[TALA+TALB] operator_minimal promoter
- 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
TAL effectors (TALEs) are able to specifically bind to the DNA by recognizing base pairs on a DNA molecule. With TAL regulators we are able to control expression of a desired gene. Efficient transcription control can be achieved by binding of multiple TAL regulators upstream of promoter.
Our construct contain ten binding sites for TALA, another ten for TALB and minimal promoter. Minimal promotor is taken from pGL4 vector series (promotor is described [http://www.promega.com/~/media/files/resources/protocols/technical%20manuals/0/pgl4%20luciferase%20reporter%20vectors%20protocol.pdf?la=en here] on the page 18).
Single binding site is combined from 18 base pairs recognised by specific TAL. TALA and TALB binding sites are adjacent to each other making impossible for both of them to bind at once.
Single binding site sequence for TALA: TTTACTGCTGCTCCCGCT
Single binding site sequence for TALB: TCTTCCGTTTCCACATCT
Figure 1: Schematic representation of the construct.
- Binding sites for TAL effectors were ordered from IDT.
Characterization
Construct was later used for constructing different reporter plasmids to show specific binding and competition between TALA and TALB.
Results:
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 396
Illegal NgoMIV site found at 761
Illegal AgeI site found at 259
Illegal AgeI site found at 599
Illegal AgeI site found at 624
Illegal AgeI site found at 964 - 1000COMPATIBLE WITH RFC[1000]