Melbourne/Blue Photosensor Background

From 2007.igem.org

(Difference between revisions)
Line 1: Line 1:
[[Melbourne/Background|<return to top of background>]]  [[melbourne|<return to home page>]]   
[[Melbourne/Background|<return to top of background>]]  [[melbourne|<return to home page>]]   
-
==Preliminaries==
+
The background to the design of the blue photosensor is included here. Specifically the design for the chimeric fusion protein that will be the key link in blue light pathway
 +
 
 +
 
 +
== Preliminaries ==
 +
Restriction Site Analysis - [http://tools.neb.com/NEBcutter2/index.php|NEB cutter]
 +
Sequence Translation - [http://au.expasy.org/tools/dna.html|Translate]
 +
Sequence Aligment - [http://www.jalview.org/download.html|JalView]
 +
PCR primer design - [http://www.premierbiosoft.com/netprimer/netprlaunch/netprlaunch.html|NetPrimer]
 +
 
 +
 
 +
==Sequences and Restriction Sites==
==== SopII ====
==== SopII ====

Revision as of 12:04, 6 August 2007

<return to top of background> <return to home page>

The background to the design of the blue photosensor is included here. Specifically the design for the chimeric fusion protein that will be the key link in blue light pathway


Contents

Preliminaries

Restriction Site Analysis - cutter Sequence Translation - [1] Sequence Aligment - [2] PCR primer design - [3]


Sequences and Restriction Sites

SopII

sensory rhodopsin II

DNA, AA, genbank

  • No IGEM restriction sites
  • HaeII (@168) present


HtrII

sensory rhodopsin II transducer

DNA, AA, genbank

  • No IGEM restriction sites
  • No useful restriction site


SopII/HtrII fusion

DNA, AA (frame 2 in Translate - only SopII + HtrII + Linker included in link), linker = TSASA SNGASA,

genbank

  • No IGEM restriction sites
  • Use site from SopII (this is the one that will be used)


ComP

two-component sensor histidine kinase

DNA (includes IGEM forward and reverse primers (VF2 + VR) and IGEM prefix and suffix, AA, genbank

SpeI (@927) deleted HaeII (@1902) inserted


ComA

sensory rhodopsin II transducer

DNA, AA, genbank

SpeI (@12) deleted HaeII (@433) inserted


Chimera Design

Overview

Refer to 4 PCR primer method here, 2001 paper

SopII Analysis

transmembrane photoreceptor, requires all-trans retinal as substrate... linked to HtrII insert TM analysis here. reference the three papers cited in background

HtrII Analysis

Yeh!!!! sequence analysis.

ComP Analysis

Chosen Fusion

File:Seq align.jpg Media:Example.ogg