BBa_K2112001

BBa_K2112001 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K2112001
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Fanglin Zhao
Date created: 2016-10-13 11:00:00
Date modified: 2016-10-14 06:03:07

OP Pesticides Residue Killer



Types
DnaRegion

Roles
engineered_region

Composite

Sequences BBa_K2112001_sequence (Version 1)

Description

The plasmid could be co-transformed into E.coli BL21(DE3) with another plasmid containing a lacI coding sequence. Then with the inducing by IPTG (a compound), target protein could be expressed in high efficiency.
T7 promoter is a strong promoter to launch the transcription procedure.
Lac operator DNA sequence could bind a protein named LacI coded by a lacI DNA sequence which located in another plasmid. This process stop the transcription of the target gene (opdA). So in our study, we add a compound named IPTG to the culture medium. IPTG could bind LacI protein, so that the structure of LacI will be changed and could not bind to lac operator DNA sequence any more. And the target gene opdA could be transcribed and translated.
His-tag sequence (fused in N-terminal of target protein OpdA) consisted of five to six histidines, which could bind to Nickel metal ions. So with a special resin chelated with nickel metal ions, we can purify the expressed OpdA protein easily.
When??the??degradation??finishes,??ultraviolet??light??or??natural??light??is??given??to??shed??on??the??bacteria,??and??thus??the??RecA??repair??gene??inside??the??bacteria??is??activated.??The??product??of??this??gene??can??activate??RecA??(SOS)??promoter.??Thus??the??bacteria??begin??to??express??suicide??gene??ccdB,??which??leads??the??bacteria??to??dying??automatically??in??order??to??avoid??the??secondary??pollution??on??vegetables??and??fruits??posed??by??genetically??engineered??bacteria.

Notes

The his-tag can allow us to purify the protein after the bacteria secrete it, so that if the project can be use in real life, the posibility of risk can be limited by giving the users the purified protein to degrade organophosphorous instead of using the bacteria.

Source

The ccdB gene is found by other teams, we use it in our part to achieve the goal.

igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K2112001/1