BBa_K3994001
== Profile ==
Name: HrpR
Base Pairs: 948bp
Origin: Pseudomonas syringae, genome
Properties: A coding sequence of HrpR protein
== Usage and Biology ==
This codes for HrpR protein. HrpR protein binds to HrpS protein forming a complex and then triggering the transcription of promoter hrpL. This functions like a AND logic gate.
BBa_K3994003
== Profile ==
Name: HrpS
Base Pairs: 912bp
Origin: Pseudomonas syringae, genome
Properties: A coding sequence of HrpS protein
== Usage and Biology ==
This codes for HrpS protein. HrpR protein binds to HrpS protein forming a complex and then triggering the transcription of promoter hrpL. This functions like a AND logic gate.
BBa_K3994008
== Profile ==
Name: PyeaR_HrpR
Base Pairs: 1223 bp
Origin: Synthetic
Properties: A coding sequence of HrpR protein.
BBa_K3994009
== Profile ==
Name: PttB344_HrpS_PJ23105_ttrR
Base Pairs: 2040 bp
Origin: Synthetic
Properties: A biosensor releasing fluorescence only when detecting the IBD marker.
== Usage and Biology ==
===Background===
Inflammatory bowel disease (IBD) is a chronic intestinal inflammatory disease of unknown etiology, including ulcerative colitis (CD) and Crohn’s disease (CD). This chronic disease, which is prone to repeated deterioration, currently lacks unified diagnostic and treatment standards, and is posing a great threat to public health. Drug therapy (anti-inflammatory drugs) is the preferred treatment for IBD. However, studies in the past 10 years have found that 30-50% of IBD patients do not respond to anti-TNF treatment. In addition, after long-term use of anti-inflammatory drugs, the patient's intestinal microbial status changes over time, and the effect may be lost due to drug resistance. Therefore, we need to seek help from other treatments for IBD.
== Construct design ==
HrpS is a key functional factor under PttB344 promoter (Figure 1). And this sequence is inserted into plasmid. Its sequence is shown in Figure 2.
Figure 1. Boolean gate logic circuit/AND circuit construction.
Figure 2. PttB344_HrpS_PJ23105_ttrR, sensing S4O62- to release the substance S
BBa_K3994010
== Profile ==
Name: PhrpL_amilGFP_IL10_PJ23104_ttrS
Base Pairs: 4231 bp
Origin: Synthetic
Properties: A coding sequence of amilGFP and IL10 protein.
== Construct design ==
This part is a coding sequence of amilGFP and IL10. amilGFP is a fluorescent protein. IL10 is an anti-inflammatory factor. These two factors were under PhrpL promoter. And this part is inserted into plasmid.
Figure 1. PhrpL_amilGFP_IL10_PJ23104_ttrS box.
Figure 2. The sequence map of PhrpL_amilGFP_IL10_PJ23104_ttrS.
BBa_K3994011
== Profile ==
Name: PyeaR_HrpR+PttB344_HrpS_PJ23105_ttrR+PhrpL_amilGFP_IL10_PJ23104_ttrS
Base Pairs: 7494 bp
Origin: Synthetic
Properties: A biosensor releasing fluorescence only when detecting the IBD marker.
== Construct design ==
We can build Boolean logic circuits to combine thiosulfate/tetrasulfate and nitrate biosensors to optimize performance, improve sensor accuracy and enhance robustness, and optimize intestinal inflammation detection (Figure 1). When and only when S2O32-/S4O62- and NO3- are present at the same time, the fluorescent gene is expressed (Figure 1). The fluorescent gene can be replaced with a gene that produce the interleukin-10 (anti-inflammatory factor) for the treatment of IBD.
HrpS is a key functional factor under PttB344 promoter. amilGFP is a fluorescent protein under PhrpL promoter. IL10 is an anti-inflammatory factor.
Figure 1. Boolean gate logic circuit/AND circuit construction.
Part1: PyeaR_HrpR, sensing NO3- to release the substance R;
Part2: PttB344_HrpS_PJ23105_ttrR, sensing S4O62- to release the substance S;
Part3: PhrpL_amilGFP_IL10_PJ23104_ttrS, will release fluorescence if sensing both the substances, R and S released from Part 1(HrpR) and Part 2 (HrpS) , respectively.
B. PttB344_HrpS_PJ23105_ttrR, sensing S4O62- to release the substance S;
C. PhrpL_amilGFP_IL10_PJ23104_ttrS.
BBa_K3994012
== Profile ==
Name: PttB344
Base Pairs: 85bp
Origin: Synthetic
Properties: A coding sequence of promoter ttB344.
== Usage and Biology ==
This is a sequence of PttB344. PttrB185-269 is a minimal TtrR activated promoter when TtrR is phosphorylated by TtrS after TtrS sensing tetrathionate.