The ccdB gene is found by other teams, we use it in our part to achieve the goal.
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.
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.
Coding region for the ccdB (control of cell death) gene.
The recA(SOS)promoter gene from the chromosomal DNA isolated from MG1655 has been PCR amplified and inserted into plasmid pSB1A2(part number BBa_B0015).
Opda is a phophotriesterase from Agrobacterium that can detoxify a broad range of organophosphate pesticides and nerve agents.
All biobrick restriction sites have been removed.
Strong IPTG-inducible expression of the OpdA enzyme.
BBa_R0010 + BBa_BOO34 +BBa_K145151 + BBa_C0012 Part of a suicide system.
This part contains the promoter, rbs and terminator.
An organophosphorus hydrolase(OPH) than can degrade organophosphorus compounds
R0011+B0034, strong IPTG-inducible promoter with strong RBS.
This part contains the strong, IPTG-inducible promoter R0011, combined with the strong RBS B0034. This part can be used as a strong protein expression system when combined with a protein coding sequence.
This part should offer greater levels of expression when compared to similar parts using the natural lac promoter (R0010).
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Promoter (lacI regulated, lambda pL hybrid)
represillator of Elowitz and Leibler (2000)
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The PLlac 0-1 promoter is a hybrid regulatory region consisting of the promoter P(L) of phage lambda with the cI binding sites replaced with lacO1. The hybrid design allows for strong promotion that can nevertheless be tightly repressed by LacI, the Lac inhibitor (i.e. repressor) (<bb_part>BBa_C0010</bb_part>) ([LUTZ97]). The activity of the promoter can be regulated over a >600-fold range by IPTG in E.Coli DH5-alpha-Z1 (same paper reference).
hybrid promoter design to create strong promoter that is, at the same time, highly repressible. note that the upstream operator installed in this hybrid is slightly different than the one in the original source (Lutz and Bujard, 1997). the most upstream operator region is slightly truncated in the represillator version, so that both operators in the hybrid are the same sequence. see references for details. also, the sequence has been truncated after the transcriptional start site. LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and increase transcription. This part is incompatible with environments containing lactose or lactose analogs.
Neelaksh Varshney, Grace Kenney, Daniel Shen, Samantha Sutton
Conversion of the iGEM parts registry to SBOL2.1