Difference between revisions of "Part:BBa K365002"

Latest revision as of 18:59, 15 September 2013

Phytochrome B from A. thaliana (aa 1-908)

Note: This is a N-part RFC[25] Biobrick which means only protein fusions to the C-terminus of this Biobrick is possible. For a improved version of this Biobrick which also allow fusion to the N-terminus of PhyB (e. g. Strep-tag II or His-tag), please see here: BBa_K801031

This part consists in the first 908 amino-acids of Phytochrome B of A. thaliana. When bound to its natural chromophore Phycocyanobiline (PCB) it can reach its Pfr activated state and physically interact with different PIF.

Background:

Phytochromes characterised by a red/far-red photochromicity. Through red-light (650–670 nm) absorption the phytochrome undergoes a rapid conformational change from its ground state Pr to its active state Pfr. The structural change allows the binding of the PIF. This light-sensitive interaction has been mapped to the 650-residue amino-terminal photosensory core of PhyB (Khanna et al., 2004). The process is completely reversible through absorption in the near infra-red spectrum (705–740 nm).

The photoreceptor protein PhyB serves for the light-dependent activation of the system, therefore it will be fused to the N-teminal of the ClpX-trimer.

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Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 490
Illegal BamHI site found at 572
Illegal XhoI site found at 523
Illegal XhoI site found at 542
Illegal XhoI site found at 2677
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 2062
Illegal BsaI site found at 2468
Illegal SapI site found at 739

Conception:

In in-vivo applications it has been shown that the PIF-interaction with the PhyB photosensory core (residues 1–650) is irreversible in infrared light. Lim & Voigt (2009) demonstrated by assaying PIF6 (which has the strongest interactions of all previously reported PIF domains) against different variants of PhyB that the tandem C-terminal PAS domains (residues 1-908)of plant phytochromes are necessary to confer rapid photoreversibility under infrared light. The original sequence contains a SpeI restriction within the first 908 residues.

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K365002 short</partinfo>
+'''''Note: This is a N-part RFC[25] Biobrick which means only protein fusions to the C-terminus of this Biobrick is possible. For a improved version of this Biobrick which also allow fusion to the N-terminus of PhyB (e.&nbsp;g. ''Strep''-tag II or His-tag), please see here: [https://parts.igem.org/Part:BBa_K801031 BBa_K801031]'''''
 This part consists in the first 908 amino-acids of Phytochrome B of A. thaliana. When bound to its natural chromophore Phycocyanobiline (PCB) it can reach its Pfr activated state and physically interact with different PIF.