SacB-Human Elastin (EP20-24-24) Fusion Protein
Elastin is a polymeric extracellular matrix protein found in tissues that require the ability to extend and recoil. Examples of elastin containing tissues include arteries, lungs, ligaments and skin. Construct EP20-24-24 for human elastin polypeptide consists of distinct exons which code for alternating hydrophobic regions and crosslinking domains from the human elastin polypeptide gene 1. Under appropriate conditions of temperature and ionic strength, elastin polypeptide undergoes a self-aggregation process known as coacervation. Coacervation is usually induced by an increase in temperature and causes the protein to separate from the solution as a second phase. Unlike most proteins which undergo denaturation when the temperature of the solution increases, elastin polypeptides become more ordered through coacervation 2. SacB is a signal peptide used in the Sec-SRP (secretory signal recognition particle) pathway by ''B. subtilis''. Signal peptides are responsible for directing preproteins (secretory proteins with a signal peptide region attached) through an appropriate secretory pathway. In the case of the Sec-SRP signal peptide, they direct preproteins from the cytoplasm into the growth medium. SacB has been successfully used in the secretion of heterologous proteins such as acid-stable α-amylase, cystatin and interleukin-3 by B.subtilis 3.
BioBrick standard was applied to the SacB-Human Elastin(EP20-24-24) Fusion Protein.
All exons in EP20-24-24 are derived from the human elastin polypeptide gene. EP20-24-24 was used to study the effect of various combinations of exons on coacervation of elastin polypeptide. SacB was identified from the initial part of certain preprotein genes that utilises the the Sec-SRP secretory pathway 4. Both components were synthesised as a fusin protein by GeneArt.
Hair-pin structure can protect antisense FtsZ.
LipA-Human Elastin (EP20-24-24) Fusion Protein
Elastin is a polymeric extracellular matrix protein found in tissues that require the ability to extend and recoil. Examples of elastin containing tissues include arteries, lungs, ligaments and skin. Construct EP20-24-24 for human elastin polypeptide consists of distinct exons which code for alternating hydrophobic regions and crosslinking domains from the human elastin polypeptide gene 1. Under appropriate conditions of temperature and ionic strength, elastin polypeptide undergoes a self-aggregation process known as coacervation. Coacervation is usually induced by an increase in temperature and causes the protein to separate from the solution as a second phase. Unlike most proteins which undergo denaturation when the temperature of the solution increases, elastin polypeptides become more ordered through coacervation 2. LipA is a signal peptide from the B.subtilis genome. In general, signal peptides are responsible for directing preproteins (secretory proteins with a signal peptide region attached)through an appropriate secretory pathway3. LipA has been successfully used in the secretion of heterologous proteins such as cutinase by ''B. subtilis''.
BioBrick standard was applied to the LipA-Human Elastin(EP20-24-24) Fusion Protein.
All exons in EP20-24-24 are derived from the human elastin polypeptide gene. EP20-24-24 was used to study the effect of various combinations of exons on coacervation of elastin polypeptide. LipA originated from the B. subtilis genome4. Both components were produced as a fusion protein by GeneArt.
p70-CsgA-His*2, double His-tagged curli generator
Juliette Paillet, Alexandre Duprey, Emiel van der Kouwe
EAK16-II is a sixteen amino acid peptide that self-assembles to form β-sheet structures in an aqueous medium. The alternating positive and negative charges (--++--++) are responsible for creating an electrostatic attraction between adjacent peptides 1, triggering self-assembly when the EAK16-II peptides are exposed to physiological media or salt solution. When examined under SEM, a well-ordered nanofibre structure is formed by the association of the EAK16-II peptides and these nanofibres can futher aggregate to form a membranous 3D scaffold. SacB is a signal peptide used in the Sec-SRP (secretory signal recognition particle) pathway by ''B. subtilis''. Signal peptides are responsible for directing preproteins (secretory proteins with a signal peptide region attached) through an appropriate secretory pathway. In the case of the Sec-SRP signal peptide, they direct preproteins from the cytoplasm into the growth medium. SacB has been successfully used in the secretion of heterologous proteins such as acid-stable α-amylase, cystatin and interleukin-3 by B.subtilis 2.
BioBrick standard was applied to the SacB-EAK16-II Fusion Protein.
EAK16-II is a segment from zuotin, a yeast protein. SacB was identified from the initial part of certain preprotein genes that utilises the the Sec-SRP secretory pathway 3. Both components were synthesised as a fusin protein by GeneArt.
expression PⅢ, which is from M13 phage genome, with the regulation of araBAD promoter.
p70-CsgA-His, His-tagged curli generator
LsrA of LuxS/AI-2 signaling pathway in Salmonalla
In a process called quorum sensing, bacteria communicate with one another using secreted chemical signaling molecules termed autoinducers. As the density of the bacterial population increases, so does the concentration of secreted autoinducer molecules. When the concentration of the autoinducer reaches a critical threshold, it diffuses back into the cell and activates or represses certain target genes. An autoinducer called AI-2 is generated by many species of Gram-negative and Gram-positive bacteria. In the LuxS/AI-2 signaling system of Salmonella enterica serovar Typhimurium, AI-2 response involves genes named lsr (LuxS regulated) that encode ATP binding cassette-type transporter. And LsrA is ATP-binding protein of the ABC transporter. In our project, we set this part under a nisA promoter for the final purpose of constructing an integrated AI-2 response pathway in our engineering bacteria.
The lsrA gene was isolated from the genome of Salmonella enterica subsp. enterica serovar Typhimurium str. LT2 using PCR.We added restriction enzyme sites when designing primers,and the assembly is done through the use of restriction sites (cutting and ligating).
It comes from the genome of Salmonella enterica subsp. enterica serovar Typhimurium str. LT2(Gene ID: 1255601)
EAK16-II is a sixteen amino acid peptide that self-assembles to form β-sheet structures in an aqueous medium. The alternating positive and negative charges (--++--++) are responsible for creating an electrostatic attraction between adjacent peptides 1, triggering self-assembly when the EAK16-II peptides are exposed to physiological media or salt solution. When examined under SEM, a well-ordered nanofibre structure is formed by the association of the EAK16-II peptides and these nanofibres can futher aggregate to form a membranous 3D scaffold. LipA is a signal peptide from the B.subtilis genome. In general, signal peptides are responsible for directing preproteins (secretory proteins with a signal peptide region attached)through an appropriate secretory pathway2. LipA has been successfully used in the secretion of heterologous proteins such as cutinase by ''B. subtilis''.
BioBrick standard was applied to LipA-EAK16II Fusion Protein.
EAK16-II was identified as a region in zuotin, a Z-DNA binding protein from the yeast genome. LipA originated from the B. subtilis genome3. Both components were produced as a fusion protein by GeneArt.