Difference between revisions of "Part:BBa K896001"

Revision as of 16:58, 26 September 2012

CysI (Sulfite reductase)

Our engingeered cyanobacteria can remove Sulfur Oxides (SOX, SO2),main precursors of air pollution and produce H2S for futher uses(Sulfide-Quinone Reductase,sqr).

CysI( surfur reductase) :This is a enzyme that can reduce (HSO3)- to H2S using the embedded electron transfer system in pseudomonas aeruginosa PAOI.

[http://www.rcsb.org/pdb/explore/explore.do?structureId=3MM5 sulfite reductase in complex with the substrate sulfite]

Usage and Biology

1. From clinical research, we fund out that Pseudomonas aeruginosa can produce great amount H2S.So, we logical assumed there are some pivotal enzymes can transfer (HSO3)- to H2S.

2. We searched for [http://www.genome.jp/kegg/ KEGG] and [http://www.ncbi.nlm.nih.gov/ NCBI] and found out CysI, a sulfite reductase inside [http://www.ncbi.nlm.nih.gov/protein/NP_250529.1 Pseudomonas aeruginosa PAOI].

3. We've already constructed CysI gene into E.coli and cyanobacteria and used bacteria to remove SO2 in our environment. Also, this bacteria produced H2S, which is a substrate for Sulfide-Quinone Reductase(sqr).

4.These potential CysI transformed bacteria can perform bioremediation, also are a powerful species for Veniusian!!

gene construction & cloning

1.we get the whole gene sequence of Cys I from NCBI web (http://www.ncbi.nlm.nih.gov/gene/878581).

2.Cys I contain endogenous EcoRI and PstI site, so we can’t clone into PSB1C3 directly.As a result, We create a noval cassette- new pSB1C3( Mfe I-Xba I -pSB1C3-Sbf I-Spe I) for easily cloning.

3.Taking advantage of MfeI and EcoRI are compatible, also SbfI and XbaI, we can easily clone Cys I gene into pSB1C3 standard biobrick.

4.Enzyme check by XbaI and SbfI, we can get ~1700bp band, which means our gene constraction is correct!!

A powerful cassette for CysI engingeering, you can use customerized promoter as you want!!

Sequence and Features

Functional Parameters