Team:Shanghaitech/Library
Collaborations：Library
This year, we have constructed several sensors and reporters as our inputs and executors to provide users more options when using the MagicBlock system. These blocks include lacI-lasI, which generates signals upon induction; pcons+GFP, which constitutively generates signals; lasR-GFP and lasR-mRFP, which present different fluorescence and lasR-amilCP, which gives bacteria resistance to chloramphenicol. Meanwhile, we also collected parts that can be reformed for our MagicBlock from other teams. For example, we have collected promoters that can sense nitrogen from UCAS, an auto-inducer quorum-sensing generator from SiCAU-China, a toolbox of recombinase from Fudan-China, cellulase and pectinase from LanZhou iGEM team, and Bt toxin protein from FAFU-China. In addition to part collection, we also reconstructed some MagicBlocks from previous characterized parts so that they can be integrated into our project. Thus our library was and will be continuously improved by users to increase the versatility.
| name | Team | Function Description | experience | |
|---|---|---|---|---|
| glnAp2+riboJ+RBS | UCAS | ucasigem@163.com | A sigma54-dependent promoter together with its regulating region, responsing to the changes of nitrogen concentration in the environment. Originally regulate the transcription of glnA. | Part:BBa_K2287001 |
| glnHp2 | UCAS | ucasigem@163.com | A sigma54-dependent promoter together with its regulating region, responsing to the changes of nitrogen concentration in the environment. Originally regulate the transcription of glnH. | Part:BBa_K2287002 |
| astCp | UCAS | ucasigem@163.com | A sigma54-dependent promoter together with its regulating region, responsing to the changes of nitrogen concentration in the environment. Originally regulate the transcription of astC. | Part:BBa_K2287003 |
| celluse | Lanzhou | 347684025@qq.com | celluse | Part:BBa_K2377002 |
| pectinase | Lanzhou | 347684025@qq.com | pectinase | Part:BBa_K2377003 |
| φBT1 integrase | Fudan-China | igem@fudan.edu.cn | This ORF codes for the steptomyces phage φBT1 integrase, which catalyzes a site specific recombination between φBT1 attB and attP sequences. | Part:BBa_K2460001 |
| φBT1 attB | Fudan-China | igem@fudan.edu.cn | the sequence which can be recoginized by the steptomyces phage φBT1 integrase | Part:BBa_K2460002 |
| φBT1 attP | Fudan-China | igem@fudan.edu.cn | the sequence which can be recoginized by the steptomyces phage φBT1 integrase | Part:BBa_K2460003 |
| φRv1 integrase | Fudan-China | igem@fudan.edu.cn | This ORF codes for the mycobacterium phage φRv1 integrase, which catalyzes a site specific recombination between φRv1 attB and attP sequences. | Part:BBa_K2460004 |
| φRv1 attB | Fudan-China | igem@fudan.edu.cn | the sequence which can be recoginized by the steptomyces phage φRv1 integrase | Part:BBa_K2460005 |
| φRv1 attP | Fudan-China | igem@fudan.edu.cn | the sequence which can be recoginized by the steptomyces phage φRv1 integrase | Part:BBa_K2460006 |
| TG1 integrase | Fudan-China | igem@fudan.edu.cn | This ORF codes for the steptomyces phage TG1 integrase, which catalyzes a site specific recombination between TG1 attB and attP sequences. | Part:BBa_K2460007 |
| TG1 attB | Fudan-China | igem@fudan.edu.cn | the sequence which can be recoginized by the steptomyces phage TG1 integrase | Part:BBa_K2460008 |
| TG1 attP | Fudan-China | igem@fudan.edu.cn | the sequence which can be recoginized by the steptomyces phage TG1 integrase | Part:BBa_K2460009 |
| 4A5-R-IG(△LVA) | SiCAU-China | linjj23@126.com | It's a positive feedback system, which could appear strong green fluorescence by background expression in BL21(DE3) or other host. | Part:BBa_K2311002 |
| 4A5-R-IG(△LVA)-AiiA | SiCAU-China | linjj23@126.com | It's a positive feedback system, which could appear strong green fluorescence by background expression in BL21(DE3) or other host. But this system may need more time to show postive feedback effect than the positive feedback system without AiiA control | Part:BBa_K2311001 |
| Cry4Aa4 + Extended FMDV | FAFU-China | jacarandasmile@163.com | Cry4Aa is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex by bioassay. The Cry protein is consisted of three functional domains. Domain I is a seven α-helices bundle. It can insert itself into a membrane by using its hydrophobic helices α4 and α5 to insert into the phospholipid bilayer. The pore formation occurs on its α3 helix. Domain II and domain III are two β-sheets which are involved in the receptor interactions. Domain II contains extremely variable loops, which are the binding site of the receptor. Domain III has the function of stabilizing the toxin. Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer ，Then the oligomer binds to a GPI-anchored APN or ALP. Secondly, the previous binding induces the oligomer insertion into the lipid rafts membrane. A formation of ion permeable pore is followed by the insertion which allows small molecules to pass through the membrane. The membrane potential inevitably changes greatly, causing the swelling of cell and finally breaking down. When the cell lysis reaches to a certain degree, the midgut necrosis and epithelial denaturation follow. Then, the alkaline hypertonic inclusions in midgut enters into hemocoel and the pH of haemolymph rises causing paralysis of larvae and finally death. To increase the express level in Chlamydomonas reintmrdtii, we synthesis Chlamydomonas reintmrdtii codon optimized Cyt1. Meanwhile, we added 2A peptide sequence at the end of 5’ According to the published paper and the result of Swiss-model, there is no effect to the toxicity of Cry or Cyt. Users can use infusion technology to link the express vector to express toxins | Part:BBa_K2074021 |
| Cry10Aa4 + Extended FMDV | FAFU-China | jacarandasmile@163.com | Cry10Aa is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex and Aedes by bioassay. The Cry protein is consisted of three functional domains. Domain I is a seven α-helices bundle. It can insert itself into a membrane by using its hydrophobic helices α4 and α5 to insert into the phospholipid bilayer. The pore formation occurs on its α3 helix. Domain II and domain III are two β-sheets which are involved in the receptor interactions. Domain II contains extremely variable loops, which are the binding site of the receptor. Domain III has the function of stabilizing the toxin. Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer. | Part:BBa_K2074022 |
| cyt1 | FAFU-China | jacarandasmile@163.com | Cyt1 is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex and Aedes by bioassay. Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer，Then the oligomer binds to a GPI-anchored APN or ALP. Secondly, the previous binding induces the oligomer insertion into the lipid rafts membrane. A formation of ion permeable pore is followed by the insertion which allows small molecules to pass through the membrane. The membrane potential inevitably changes greatly, causing the swelling of cell and finally breaking down. When the cell lysis reaches to a certain degree, the midgut necrosis and epithelial denaturation follow. Then, the alkaline hypertonic inclusions in midgut enters into hemocoel and the pH of haemolymph rises causing paralysis of larvae and finally death. To increase the express level in Chlamydomonas reintmrdtii, we synthesis Chlamydomonas reintmrdtii codon optimized Cyt1. Meanwhile, we added 2A peptide sequence at the end of 5’ According to the published paper and the result of Swiss-model, there is no effect to the toxicity of Cry or Cyt. Users can use infusion technology to link the express vector to express toxins. | Part:BBa_K2074024 |
| cyt2 | FAFU-China | jacarandasmile@163.com | Cyt2 is cloned from Bacillus thuringiensis BRC-LLP29. It shows specific toxicity to Culex and Aedes by bioassay.Cyt proteins have a single α-β domain which do not bind to receptors but can directly insert into the cell membrane and then form a pore causing cell death. Although Cry and Cyt proteins are two big families of δ-endotoxins, they are far related.Cyt1 and Cyt2 are two types of Cyt proteins found in Bti.. Generally, Cry proteins are believed to exert toxicity by interacting with the proteins on the brush border membrane and then insert into the membrane which takes multiple steps. At the beginning in mosquitoes' gut, the crystalline inclusions are cleaved at the disulfide bond to release the Cry pre-toxin. Then the soluble proteins are activated by being cleaved again by intestinal protease. When toxins reach to the brush border membrane microvilli, they bind to the proteins, or known as receptors on the membrane. The binding process takes two step. Firstly, the monomeric Cry toxin binds to cadherin, resulting in the formation of pre-pore oligomer | Part:BBa_K2074025 |
