Team:ECNUAS/Contribution

ECNUAS

Contribution

BBa_K3995002

== Profile ==

Name: Pprovoin5
Base Pairs: 82bp
Origin: Pseudomonas sp.,genome
Properties: atzDEF/atzR operator

== Usage and Biology ==

This operator region has a leftward-facing repressible promoter and a rightward-facing activated promoter. AtzR remains bound to the DNA. In the natural system, the left-facing promoter is upstream of the atzR CDS, and atzR is autoregulatory; in the presence of cyanuric acid, the binding of AtzR changes to cause the right-facing promoter to become activated, allowing for the expression of the downstream atzDEF CDS. The presence of cyanuric acid "frees" AtzR from being bound to the atzDEF promoter so polymerase can now bind and transcribe following gene. AtzR remains bound to the atzR promoter region even in the presence of cyanuric acid. Note that high glnK concentrations are also needed, as glnK is a co-activator of downstream operator.

== Construct design ==

In our group, amilGFP is a key functional factor that show the signs of fluorescence which is controlled by a Pprovoin5 promoter (Figure 2). The Pprovoin5_amilGFP is inserted in the pUC57 mini vector (Figure 3).

Figure 2. Pprovoin5_amilGFP box.

Figure 3. Schematic maps of Pprovoin5_amilGFP (reporter plasmid).

BBa_K3995003

== Profile ==

Name: P7_AtzR
Base Pairs: 1142 bp
Origin: Synthetic
Properties: A coding sequence for atzR

== Usage and Biology ==

P7_AtzR is a coding sequence for expressing atzR. Cyanuric acid combination with AtzR can activate amilGFP expression to show the signs of fluorescence. The content of cyanuric acid is detected by detecting the fluorescence concentration. So P7_AtzR should be used with Pprovoin5_amilGFP.

Figure 1. Genetic design of our biosensor

== Construct design ==

The atzR is under T7 promoter. The P7_atzR is inserted in the pUC57 mini vector (Figure 3).

Figure 2. P7_AtzR box.

Figure 3. Schematic maps of P7_AtzR (sensor plasmid)

BBa_K3995004

== Profile ==

Name: Pprovoin5_amilGFP
Base Pairs: 976 bp
Origin: Synthetic
Properties: A coding sequence for amilGFP.

== Usage and Biology ==

Pprovoin5_amilGFP is a coding sequence for expressing amilGFP. Atrazine is one of the most widely used herbicides in the world. It is cheap and also effective on many crops. But it still has negative effects. Atrazine will leave a residue in the soil called cyanuric acid, it has endocrine disruption, genotoxicity, reproductive toxicity and other harm to mammals. Pprovoin5_amilGFP functions as a cell-free gene element to detect cyanuric acid. Cyanuric acid combination with AtzR can activate amilGFP expression to show the signs of fluorescence. The content of cyanuric acid is detected by detecting the fluorescence concentration. So the Pprovoin5_amilGFP should be used with Ptat_AtzR.

Figure 1. Genetic design of our biosensor

== Construct design ==

amilGFP is key functional factors that show the signs of fluorescence which is controlled by a Pprovoin5 promoter (Figure 2). The Pprovoin5_amilGFP is inserted in the pUC57 mini vector to get plasmid A (Figure 3).

Figure 2. Pprovoin5_amilGFP box.

Figure 3. Schematic maps of Pprovoin5_amilGFP (reporter plasmid).

BBa_K3995005

== Profile ==

Name: Ptat_AtzR
Base Pairs: 1278 bp
Origin: Synthetic
Properties: A coding sequence for atzR

== Usage and Biology ==

Ptat_AtzR is a coding sequence for expressing atzR. Cyanuric acid combination with AtzR can activate amilGFP expression to show the signs of fluorescence. The content of cyanuric acid is detected by detecting the fluorescence concentration. So Ptat_AtzR should be used with Pprovoin5_amilGFP.

== Construct design ==

The atzR is under Ptat promoter. The Ptat_atzR is inserted in the pUC57 mini vector to get plasmid A (Figure 2).

Figure 2. Ptat_AtzR box.

Figure 3. Schematic maps of Ptat_AtzR (sensor plasmid)

BBa_K3995007

== Profile ==

Name: Ptat_AtzR+Pprovoin5_amilGFP
Base Pairs: 2254 bp
Origin: Synthetic
Properties: A coding sequence for detecting cyanuric acid.

== Usage and Biology ==

Ptat_AtzR+Pprovoin5_amilGFP is a coding sequence for detecting cyanuric acid. Atrazine is one of the most widely used herbicides in the world. It is cheap and also effective on many crops. But it still has negative effects. Atrazine will leave a residue in the soil called cyanuric acid, it has endocrine disruption, genotoxicity, reproductive toxicity and other harm to mammals. Ptat_AtzR+Pprovoin5_amilGFP functions as a cell-free gene element to detect cyanuric acid. Cyanuric acid combination with AtzR can activate amilGFP expression to show the signs of fluorescence. The content of cyanuric acid is detected by detecting the fluorescence concentration.

== Construct design ==

amilGFP is key functional factors that show the signs of fluorescence which is controlled by a Pprovoin5 promoter (Figure 2). The atzR is under Ttat promoter. The Pprovoin5_amilGFP and the Ptat_atzR are inserted in the pUC57 mini vector respectively to get plasmid A and plasmid B (Figure 3).

Figure 2. Ptat_AtzR+Pprovoin5_amilGFP box.

Figure 3. Schematic maps of Ptat_AtzR (sensor plasmid) and Pprovoin5_amilGFP (reporter plasmid).