Difference between revisions of "Part:BBa K771006"

Revision as of 10:57, 29 September 2012

Membrane Anchor 6:ssDsbA-VVD-LGT

Membrane anchor 6, which consists of blue light sensor VVD(BBa_K771109) and membrane protein Lgt(BBa_K771102). Membrane Anchor 6 could aggregate with Membrane Anchor 5 [BBa_K771005] only when blue light is present.

Overview: Membrane Scaffold System

Demonstration of the whole Membrane Scaffold device. Membrane Anchor 2, 3, 4, 5 consitutively aggregate while Membrane Anchor 1 and 6 only aggregate with

constitutive cluster (Membrane Anchor 2, 3, 4, 5) when certain signal is present.

Backgroud

Blue Light Induced Aggregation

We incorporated VVD(BBa_K771109) as a blue light sensor to control the aggregation state of Membrane Scaffold system. VVD is a photoreceptor of Neurospora crassa and can form dimer only in the presence of blue light and disassociate as light is off. Mutant (C71V and N56K) is harder to dimerize in the dark and easier to disassociate under blue light compared with wildtype VVD.

When blue light is on, Membrane Anchor 5 and 6 aggregate, activating pathway involving enzymes 2, 3, 4, 5 and 6.

Characterization

We justified the blue light induced aggregation ability of Membrane Anchor 5 and 6 by recruiting Violacein pathway in our system. (See application)

Design

100px

Membrane Anchor 6 can aggregate with Membrane Anchor 6 only when blue light is on.

Application

We recruited Violacein biosynthetic pathway involving enzyme Vio A,B,C,D and E. Violacein biosynthetic pathway is a branched pathway which could lead to the producion of three final products.

Through blue light signal control, we successfully turned on a pathway which was otherwise almost shut off.

For more information, see BBa_K771201

Related Biobrick

Membrabe Anchor 1([BBa_K771001]) <br\>Membrabe Anchor 2([BBa_K771002]) <br\>Membrabe Anchor 3([BBa_K771003]) <br\>Membrabe Anchor 4([BBa_K771004]) <br\>Membrabe Anchor 5([BBa_K771005])

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 494
Illegal AgeI site found at 1339
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 960

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K771006 short</partinfo>
-Membrane anchor 6, which consists of interacting light sensor VVD and membrane protein Lgt. Membrane Anchor 6 is expected to aggregate with Membrane Anchor 5 only when blue light is present[[https://parts.igem.org/wiki/index.php?title=Part:BBa_K771005 BBa_K771005]].
-==Membrane Rudder System==
+Membrane anchor 6, which consists of blue light sensor VVD([https://parts.igem.org/wiki/index.php?title=Part:BBa_K771109 BBa_K771109]) and membrane protein Lgt([https://parts.igem.org/wiki/index.php?title=Part:BBa_K771102 BBa_K771102]). Membrane Anchor 6 could aggregate with Membrane Anchor 5 [[https://parts.igem.org/wiki/index.php?title=Part:BBa_K771005 BBa_K771005]] only when blue light is present.
-[[Image:12SJTU-MA1-MA6.png|500px|center]]
-Demonstration of the whole ''Membrane Rudder'' device.
-==light sensor==
+==Overview: Membrane Scaffold System==
-Photoreceptor of Neurospora crassa and can form dimer in the presence of blue light and disassociate as light is off. Mutant (C71V and N56K) is harder to dimerize in the dark and easier to disassociate under blue light compared with wildtype VVD
+[[Image:12SJTU-MA1-MA6.jpg|500px|center]]
+Demonstration of the whole ''Membrane Scaffold'' device. Membrane Anchor 2, 3, 4, 5 consitutively aggregate while Membrane Anchor 1 and 6 only aggregate with
-[[Image:12SJTU VVD dark state.png|right|400px|thumb|Dark state VVD dimers, from PDB ID 2PD7]]
+constitutive cluster (Membrane Anchor 2, 3, 4, 5) when certain signal is present.
-Light is an intriguing signal to regulate E.coli activity because it is easy to obtain, highly tunable and nontoxic. A metabolite-coupled light-switchable system could be quite fascinating.
+==Backgroud==
-Vivid(VVD) protein, a photoreceptor of ''Neurospora crassa can form dimer in the presence of blue light and disassociate as light is off. Besises, VVD protein belongs to the Per-Arnt-Sim(PAS) protein superfamily.
+===Blue Light Induced Aggregation===
-Compared with wildtype VVD, VVD mutant (C71V and N56K)(this part), is harder to dimerize in the dark and easier to dimerize under blue light . So this mutant is ideal as light sensor in Membrane Rudder device. The VVD construction connected to original Membrane Anchor is [https://parts.igem.org/Part:BBa_K771105 Part:BBa_K771005] and [https://parts.igem.org/Part:BBa_K771106 Part:BBa_K771006], for Membrane Anchor 5 and 6.
+We incorporated VVD([https://parts.igem.org/wiki/index.php?title=Part:BBa_K771109 BBa_K771109]) as a blue light sensor to control the aggregation state of Membrane Scaffold system. VVD is a photoreceptor of Neurospora crassa and can form dimer only in the presence of blue light and disassociate as light is off. Mutant (C71V and N56K) is harder to dimerize in the dark and easier to disassociate under blue light compared with wildtype VVD.
-Whether blue light is present or not could determine which enzymes be assembled to make their final product dominate.
+When blue light is on, Membrane Anchor 5 and 6 aggregate, activating pathway involving enzymes 2, 3, 4, 5 and 6.
+==Characterization==
+We justified the blue light induced aggregation ability of Membrane Anchor 5 and 6 by recruiting Violacein pathway in our system. (See application)
+==Design==
+[[Image:12SJTU_MA6.jpg|100px|center]]
+Membrane Anchor 6 can aggregate with Membrane Anchor 6 only when blue light is on.
+==Application==
+We recruited Violacein biosynthetic pathway involving enzyme Vio A,B,C,D and E. Violacein biosynthetic pathway is a branched pathway which could lead to the producion of three final products.
+Through blue light signal control, we successfully turned on a pathway which was otherwise almost shut off.
+For more information, see [https://parts.igem.org/wiki/index.php?title=Part:BBa_K771201 BBa_K771201]
 ===Related Biobrick===