Part:BBa_K771006

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

<br\>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

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