Team:SZU-China/Parts

BBa_K3838111	J-SHSP	Composite	A stress protein, also known as a heat shock protein, gives bacteria resistance to high temperatures and low pH
BBa_K3838222	J-BSH	Composite	Hydrolysis binding bile salts to produce free bile hydrochloric acid and amino acids
BBa_K3838789	J-FabV	Composite	By expressing FabV gene, we gave triclosan resistance to engineered bacteria and achieved non-antibiotic screening effect

Basic Parts

SZU-China iGEM 2021 team utilizes 5 kinds of new basic parts. Here is their individual function:
1. BBa_K3838666 and BBa_K3838233 can encode the protein of BSH and SOD,which are two of our target therapeutic products.
2. BBa_K3838613 can encode the tes4 protein that can promotes the release of butyric acid from the fatty acid synthase complex.
3. BBa_K3838777 encodes a protein called heat shock protein,which can Improve stress resistance of bacteria under extreme growing conditions.
4. BBa_K3838567 is a short amino acid peptide consisting of six histidine can be used as a label for affinity chromatography for protein purification.

Part Number	Name	Type	Part Description
BBa_K3838666	BSH	Basic	Hydrolysis binding bile salts to produce free bile hydrochloric acid and amino acids
BBa_K3838777	SHSP	Basic	A stress protein, also known as a heat shock protein, gives bacteria resistance to high temperatures and low pH
BBa_K3838233	SOD	Basic	Catalytic disproportionation of superoxide anion to oxygen and hydrogen peroxide，which plays an important role in the balance of oxidation and antioxidation in the human body
BBa_K3838613	TES4	Basic	Thioesterase that promotes the release of butyric acid from the fatty acid synthase complex Thioesterase that promotes the release of butyric acid from the fatty acid synthase complex
BBa_K3838567	6xHis Tag	Basic	A short amino acid peptide consisting of six histidine can be used as a label for affinity chromatography for protein purification.

Composite Parts

This year,we built 11 composite parts for our project.
BBa_K3838111,BBa_K3838222,BBa_K3838555,BBa_K3838888,BBa_K3838999,BBa_K3838789 are composites part that based on the promoter J23100,which can ensure stable and reliable protein expression and can meet long-term and reasonable drug administration needs.

At the same time,we built BBa_K3838123,BBa_K3838234,BBa_K3838345 to Verify the acid response mechanism we designed.

For safety reasons, we also constructed two sets of suicide mechanisms BBa_K3838321,BBa_K3838654 to prevent engineered bacteria from escaping into human blood and the environment.

You can click on the part number to see the details.

Part Number	Name	Type	Part Description
BBa_K3838111	J-SHSP	Composite	A stress protein, also known as a heat shock protein, gives bacteria resistance to high temperatures and low pH
BBa_K3838222	J-BSH	Composite	Hydrolysis binding bile salts to produce free bile hydrochloric acid and amino acids
BBa_K3838555	J-LL37	Composite	LL-37 is an antibacterial peptide secreted by human immune cells, which has an inhibitory effect on harmful bacteria
BBa_K3838888	J-SOD	Composite	Catalytic disproportionation of superoxide anion radicals to oxygen and hydrogen peroxide，which plays an important role in the balance of oxidation and antioxidation in the human body
BBa_K3838999	J-TES4	Composite	Thioesterase that promotes the release of butyric acid from the fatty acid synthase complexThioesterase that promotes the release of butyric acid from the fatty acid synthase complex
BBa_K3838789	J-FabV	Composite	By expressing FabV gene, we gave triclosan resistance to engineered bacteria and achieved non-antibiotic screening effect.
BBa_K3838345	YH-SOD	Composite	Catalytic disproportionation of superoxide anion radicals to oxygen and hydrogen peroxide，which plays an important role in the balance of oxidation and antioxidation in the human body
BBa_K3838321	phoKiller	Composite	Allowing bacteria to sense phosphate concentrations in the environment to act on the kill switch mechanism
BBa_K3838654	GluKiller	Composite	Allowing bacteria to sense glucose levels in the environment to act on the kill switch mechanism

Characterization of Existing Parts

SZU-China team has added quantitative experimental characterization data to an existing Part from the Registry of Standard Biological Parts and documented the experimental characterization on the Part's Main Page on the Registry.

This year,We identified a glucose-sensitive promoter pt-αcrp(BBa_K3142012) that can express subsequent genes at low glucose levels to prevent bacteria from escaping to the outside environment. We added the mazF gene, which encodes an endoribonuclease that will be expressed to suicidal effect when glucose levels are low. We verified our system by inducing expression with different glucose concentrations and observing the survival rate of engineered bacteria and supplemented experimental data for the promoter.

According to the literature survey, we believe that antibacterial peptide LL37 can resist the invasion of endotoxin to a certain extent. We found Part BBa_K875009 expressing the antibacterial peptide,and used this Part to express LL37 in the engineering bacteria. We carried out different levels of verification. At the same time, in order to verify whether it can effectively relieve inflammatory symptoms, we used THP-1 cell lines with and without LPS to interact with engineering bacteria, and detected the content of interleukin-6 and interleukin-10 in cells, to test whether it has the effect of slow release of inflammatory factors. The results of cell experiment can be used as the data supplement of this part.

You can click on the part number to see the details.

Part Number	Name	Type	Part Description
BBa_K3142012	PT-αcrp	Basic	Glucose starvation promoter, in response to glucose starvation
BBa_K875009	LL37	Basic	Human antimicrobial peptides which has a broad spectrum of inhibitory effects on bacteria and can bind to LPS

Improved Part

This year, our team has improved the existing part by adding the signal peptide. The improved part (J-SOD) and the original existing part (SOD) are presented below. You can click on the part number to see the details.

Part Number	Name	Type	Part Description
BBa_K3838888	J-SOD	Composite	Catalytic disproportionation of superoxide anion radicals to oxygen and hydrogen peroxide，which plays an important role in the balance of oxidation and antioxidation in the human body
BBa_M36071	SOD	Basic	encodes the SOD protein