Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Michel Saremi
Date created: 2012-09-21 11:00:00
Date modified: 2015-05-08 01:13:14
LuxCDABE
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K770001_sequence (Version 1) |
Description
Comprised of 5 Genes:-LuxA and LuxB codes respectively 40kDA and 37 kDA polypeptides, which together make up the heterodimer bacterial luciferase.
-Luciferease requires fatty aldehydes, molecular oxygen -O2 and reduced flavin mononucleotide -FMNH2 as substrates to produce blue-green light emission at 490nm, the reaction products are fatty acid, H2O and flavin mononucleotide -FMN.
Lux D: encodes transferase wich transfers Acyl-doners from the fatty acid synthetic pathway by linkage at its serine hydroxyl groups,through hydrolysis, transforming Acyl-doners to a fatty acid.
Lux E: encodes Synthetase which with ATP converts the fatty acid to a Acyl-AMP intermediate, which is highly reactive with the cysteine of Synthetase, forming an ester-thiol bond, releasing AMP.
Lux C: encodes reductase, which breaks the thiol-ester bond with NADPH to release Fatty Aldehyde.
-Together the Lux C, D and E: encodes a Fatty Acid Reductase Enzyme complex which converts acyl-doners to fatty acid then fatty aldehydes.
Notes
Contained an illegal Xbal site that had to be removed, we did so by introducing a USER-site over the restriction site containing a point mutation, this devided our LuxCDABE in two pieces of aproxymately 2 kbp and 4 kbp, which we assembled by USER-cloning with a pSB1C3 backbone, this procedure has flanked our LuxCDABE with USER-scars located between the cassette itself and our biobrick required prefix and suffix.This should not affect the transcription of the cassette itself.
The sequence of the scars and their sequence are given below.
Prefix-AGTGCGAT-LuxCDABE-ACTTGCGT-Suffix