Part:BBa_K896001

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!!

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 655
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 655
  Illegal PstI site found at 121
  Illegal NgoMIV site found at 394
  Illegal NgoMIV site found at 580
  Illegal NgoMIV site found at 722
  Illegal AgeI site found at 94
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 130
  Illegal BsaI site found at 1452
  

Functional Parameters

kegg	Pseudomonas aeruginosa PAO1: PA1838