Granulysin, a T Cell Product,Kills Bacteria by Altering Membrane Permeability
Granulysin, a protein located in the acidic granules of human NK cells and cytotoxic T cells, has microbial activity against a broad spectrum of microbial pathogens. The substance released by cytotoxic T cells (CD8) when attached to infected body cells. It creates holes in the target cell membrane and destroy it. That is,Granulysin induces apoptosis in target cells and have antimicrobial action. It is broadly antimicrobial, killing microbes that cause, for example, tuberculosis and malaria, and can destroy some tumors. A series of peptides generated from the amino acid sequence of granulysin are potential antibiotics. Granulysin increased the permeability of bacterial membranes, as judged by its ability to allow access of cytosolic ??-galactosidase to its impermeant substrate.Granulysin functions to create holes in the target cell membrane and destroy it. Granulysin is able to induce apoptosis in target cells and also has antimicrobial action.
Granulysin can be purified via nickel affinity chromatography.
Granulysin is a member of the saposin-like protein (SAPLIP) family. The gene was synthesized at GENEART in pMA vector
CAP-Bac8c, an improved antimicrobial peptide selectively
The CAP is a short peptide derived from the competence stimulation peptide(CSP) which can binding to the receptors on the membrane of the S.mutans selectively. The Bac8c is the strong antimicrobial peptide which can kill the bacterial effectively. The part is a new improved peptide ,based on the fusion of a S.mutans-specific targeting peptide domain with a wide-spectrum antimicrobial peptide domain ,which can restrain even kill the S.mutans specifically.
To reduce the effect of the CAP on the Bac8c, the CAP only has the least effective sequence of the CSP. Besides, we design an intersequence ,GGG, put between the CAP and Bac8c for the same goal.
EcAMP-1 antimicrobial peptide from barnyard grass (Echinochloa crus-galli)
RFC 23 compatible. Contains ATG (methionine).
Codon optimized for E. Coli, contains ATG (met).
pBad/araC-RBS B0034-Ag43 thanatin as alpha domain-double terminator
Ag43 is a kind of outer membrane protein that is composed of 3 domains. First one is signal peptide that transport Ag43 to outer membrane. Second one is alpha domain that has connection peptide to another Ag43 alpha domain. Last one is beta domain that has beta barrel structure to let the barrel there. This part has thanatin, a kind of antimicrobial peptide as alpha domain. This thanatin has Asp as the last residue. So if thanatin domain is digested by Asp-N endoproteinase (NEB), this part secret thanatins.
This part has 6 PstI sites so you shoud not digest this part by PstI.
This part is derived from BBa_K759001 and it contains thanatin-Asp fragment that is synthesized by IDT.
Lac operator+RBS+Signal peptide+Signiferin+Terminator
Antimicrobial peptides (AMPs) are stable peptides that have extensive ability to kill or inhibit the growth of bacteria. They play a role in defense mechanism for all organisms, ranging from prokaryotes to humans, to against predators. Some of them act against specific strain of microbes. Signiferin is a kinds of antimicrobial peptide extracted from the skin mucus of Crinia signifera, respectively. It demonstrated effectiveness in killing MRSA. And we had change it's sequence into E.coli codon usage. We treat a signal peptide in front of signiferin. This signal peptide comes from S.lividas and has ability to let E.coli secrect signiferin to LB culture medium. We used Lac operator,RBS,terminator on pQE60 to made our expression machine worked.
Signiferin with signal peptide is artificial synthesis.
This part codes for the peptide 'Signiferin' tagged with 'His-6-SUMO' molecule. Its production is triggered by IPTG (through pT7 promoter).
OH-CATH (3-34) antimicrobial peptide from the King Cobra (Ophiophagus hannah)
An antimicrobial peptide from the Cathelicidin family of peptides, isolated from the King Cobra (Ophiophagus hannah). Shortened version of the full 34 amino acid protein (C-terminal KR residues removed) that retains antibacterial properties while significantly removing hemolytic activity.
RFC 23. ATG(Met) at beginning of coding region.
Subtilosin coding region without its operon
Genomic sequence of Bacillus subtilis 168
The antimicrobial peptide, APP2, can achieve high activity exclusively at low environmental pH to target bacterial species like Streptococcus mutans that produce acid and thrive under the low pH conditions detrimental for tooth integrity. However, at pH 7.5, it has no effect on cell viability. There are only six histidine(His) and eight phenylalanine(Phe) residues present in the APP2. The pH-responsiveness of APP2 has been attributed to the presence of multiple His which enable the conformational flexibility necessary to transition from the less effective more general membrane disruptive α-helical form at close to neutral pH to its substantially more active random coil conformation at low pH. The hydrophobicity necessary for membrane interaction is provided by Phe groups which in contrast to other hydrophobic amino acids allow for the conformational flexibility needed for the pH-dependent antimicrobial activity of APP2.
Scygonadin antimicrobial peptide from the horseshoe crab (Scylla serrata)
RFC 23. ATG(Met) added to start of coding region.
Synthetic 22-amino-acid antimicrobial peptide(AMP) pexiganan, which is a magainin AMP analog isolated from the skin of the African clawed frog.
This part contains an additional BamHI restriction site at beginning of DAMP-Pex sequence and a XhoI restriction site between DAMP-Pex and double terminator(B0010-B0012). Also have a TEV Cleaveage site(ENLYFQS) between DAMP and Pex.
Antimicrobial peptides (AMPs) are stable peptides that have extensive ability to kill or inhibit the growth of bacteria. They play a role in defense mechanism for all organisms, ranging from prokaryotes to humans, to against predators. Some of them act against specific strain of microbes. Epinecidin-1 is a kind of antimicrobial peptide. This AMP is extracted from the skin mucus of Epinephelus coioides, respectively. Epinecidin-1 has been shown to help wound healing.We treat a signal peptide in front of Epinecidin-1. This signal peptide comes from S.lividas and has ability to let E.coli secrect signiferin to LB culture medium.
Epinecidin-1 is comes from Epinephelus coioides
The antimicrobial peptide, Bac8c, has a potential clinical application in preventing and treating dental caries. S.mutans treated with Bac8c shows abundance effect on bacterial structure. There appears to be a large amount of extracellular debris and obvious holes on the cell surface, as well as loss of cell wall and nucleoid condensation. What???s more, Bac8c can remarkably reduce the viability of cells in biofilms formed in the BioFlux system.
RBS - Antifungal D4E1 - Double terminator
D4E1 is a linear synthetic peptide of 17 amino acids which has shown to have antifungal activities by complexing with a sterol present in conidial wall of a variety of fungi. This part is composed of a RBS (K780002 spoVG), the open reading frame of D4E1 and a double terminator (B0015). This part was optimized for the expression and its secretion in Bacillus subtilis.
The coding sequence of D4E1 is flanked on the N-terminal end with a signal peptide (amyE signal peptide) followed by a pro peptide, cleaved during the secretion process.
De Lucca AJ,??Bland JM,??Grimm C,??Jacks TJ,??Cary JW,??Jaynes JM,??Cleveland TE,??Walsh TJ. Fungicidal properties, sterol binding, and proteolytic resistance of the synthetic peptide D4E1. Can J Microbiol.??1998 Jun;44(6):514-20.
Lactotransferrin is a major iron-binding and multifunctional protein found in exocrine fluids such as breast milk and mucosal secretions. Has antimicrobial activity, which depends on the extracellular cation concentration. Antimicrobial properties include bacteriostasis, which is related to its ability to sequester free iron and thus inhibit microbial growth, as well as direct bactericidal properties leading to the release of lipopolysaccharides from the bacterial outer membrane. Can also prevent bacterial biofilm development in P.aeruginosa infection. Has weak antifungal activity against C.albicans. Has anabolic, differentiating and anti-apoptotic effects on osteoblasts and can also inhibit osteoclastogenesis, possibly playing a role in the regulation of bone growth. Promotes binding of species C adenoviruses to epithelial cells, promoting adenovirus infection. Can inhibit papillomavirus infections. Stimulates the TLR4 signaling pathway leading to NF-kappa-B activation and subsequent pro-inflammatory cytokine production while also interfering with the lipopolysaccharide (LPS)-stimulated TLR4 signaling. Inhibits neutrophil granulocyte migration to sites of apoptosis, when secreted by apoptotic cells. Stimulates VEGFA-mediated endothelial cell migration and proliferation. Binds heparin, chondroitin sulfate and possibly other glycosaminoglycans (GAGs). Also binds specifically to pneumococcal surface protein A (pspA), the lipid A portion of bacterial lipopolysaccharide (LPS), lysozyme and DNA. Lactoferricin binds to the bacterial surface and is crucial for the bactericidal functions. Has some antiviral activity against papillomavirus infection. N-terminal region shows strong antifungal activity against C.albicans. Contains two BBXB heparin-binding consensus sequences that appear to form the predominate functional GAG-binding site.
Due to its large sequence, we had to designed in two different gBlocks.
This part was fully synthetized according to the LTF protein (UniprotKB P02788) or/and the corresponding genomic region (GenBank M73700.1; GI:619784)
Artilysin Art-E, an antibacterial fusion protein
This partcodes for their Magainin 1 antimicrobial peptide. It includes a SUMO fusion partner to allow soluble, non-toxic expression of the gene in E. coli.
Because Magainin 1 is normally cytotoxic to E. coli and other bacteria, the SUMO fusion partner is critical to successful expression of the construct. Without SUMO, the antimicrobial peptide would likely lyse the cell membrane.
The sequence for Magainin 1 was generated by reverse translating the amino acid sequence for the peptide and then codon optimising it. This gene was then synthesised. The SUMO protein fusion partner was derived from the SUMO biobrick introduced by TU Delft 2013.
T5+Lac Operator+RBS+signal peptide+Epinecidin-1+Terminater
Antimicrobial peptides (AMPs) are stable peptides that have extensive ability to kill or inhibit the growth of bacteria. They play a role in defense mechanism for all organisms, ranging from prokaryotes to humans, to against predators. Some of them act against specific strain of microbes. Epinecidin-1 is a kind of antimicrobial peptide. This AMP is extracted from the skin mucus of Epinephelus coioides, respectively. Epinecidin-1 has been shown to help wound healing.We treat a signal peptide in front of Epinecidin-1. This signal peptide comes from S.lividas and has ability to let E.coli secrect signiferin to LB culture medium. We use promoter,Lac operator,RBS,terminator on pQE 60 vector.
Cg-Defh1 antimicrobial peptide from the Pacific oyster (Crassostrea gigas)
Artilysin Art-175 fused at N-terminal with DsbA signal peptide
Artilysins are an exciting class of enzyme-based antibacterials. Their name is derived from "artificial endolysin" and they exploit the lytic power of bacteriophage-encoded endolysins. Endolysins are peptidoglycan hydrolases produced at the end of the lytic cycle that pass through the cytoplasmic membrane, degrade the peptidoglycan layer and cause the osmotic lysis of the infected bacterial cell, thus liberating the progeny. Purified endolysins have been used to kill Gram-positive pathogens. Gram-negative bacteria, however, have a protective outer membrane containing lipopolysaccharide (LPS) that serves as a barrier against the peptidoglycan hydrolytic activity of endolysins from the outside. To overcome this problem, selected polycationic or amphipathic peptides that locally destabilize the LPS layer have been covalently fused to endolysins. Biers et al. fused the sheep myeloid antimicrobial peptide (SMAP-29) to the N terminus of the endolysin KZ144 to create Artilysin Art-175. Art-175 has been shown to be a highly potent antibacterial that acts in minutes to kill virtually all Pseudomonas aeruginosa strains. [2] BBa_K1659002 is a composite of Art-175 with the N-terminal DsbA signal peptide with a hexahistidine tag. DsbA targets recombinant proteins to the co-translational signal- recognition-particle (SRP)-dependent pathway to mediate secretion out of the bacterial chassis. [2]
Endolysin KZ144 originates from Pseudomonas aeruginosa bacteriophage varphiKZ. SMAP-29 originates from sheep genomic DNA.
Artilysin Art-175 fused at N-terminal with porin-dependent YebF secretion signal protein
T5+Lac Operator+RBS+Signal peptide+Signiferin+Terminater
Antimicrobial peptides (AMPs) are stable peptides that have extensive ability to kill or inhibit the growth of bacteria. They play a role in defense mechanism for all organisms, ranging from prokaryotes to humans, to against predators. Some of them act against specific strain of microbes. Signiferin is a kinds of antimicrobial peptide extracted from the skin mucus of Crinia signifera, respectively. It demonstrated effectiveness in killing MRSA. And we had change it's sequence into E.coli codon usage. We treat a signal peptide in front of signiferin. This signal peptide comes from S.lividas and has ability to let E.coli secrect signiferin to LB culture medium. We used promoter,Lac operator,RBS,terminator on pQE60 to made our expression machine worked.
Surfactin generator for B. subtilis
In B. subtilis strains this part can induce the synthesis of surfactin, a lipopeptide with an antimicrobial activity and a powerful surfactant.
This part contains an Bacillus xylose inducible promoter, a RBS for Bacillus and the sfp gene. It can be used to induce surfactin production in Bacillus subtilis strains. However, the sfp gene does not code for the surfactin; it codes for an enzyme which catalyzes the synthesis of the lipoprotein surfactin. In order to overinduce the sfp production and therefore the surfactin synthesis in B. subtilis strains the culture media should be supplemented with xylose.
The sfp gene was isolated from many B. subtilis strains (for example ATCC 21332).
Subtilin is a lanthionine-containing peptide antibiotic (lantibiotic) active on Gram-positive bacteria. The bactericidal activity of lantibiotics is based on depolarization of energized bacterial cytoplasmic membranes, initiated by the formation of aqueous transmembrane pores. This BioBrick differs in its overhangs from BBa_K1351012.
This part codes for the peptide 'Magainin II' tagged with 'His-6-SUMO' molecule. Its production is triggered by IPTG (through pT7 promoter).
Melittin Optimized for E. coli with Mcherry
Melittin is a antimicrobial peptide found in bee venom that works to cause cell lysis by forming a pore in the membrane of harmful bacteria. While the 2014 Stony Brook iGEM team was able to successful clone the melittin gene into E. coli, expression was inconsistent. The 2015 Stony Brook iGEM team worked to correct this by optimizing the codons for an E. coli cell. Both the optimized and regular versions of melittin were fused with a mCherry tag in order to quantify differences in expression. With the use of a fluorescence plate reader, a near 100 fold increase in expression for the optimized melittin construct was observed.
While the amino acid sequence for melittin was derived from Apis mellifica, the genomic sequence was altered by hand for optimization in E. coli. The mCherry sequence was obtained from
LL-37 antimicrobial peptide from Humans (Homo sapiens)
This is an antimicrobial peptide (AMP) is commonly found in humans. This part was commercially synthesized and codon optimized for expression in E. coli and is an improvement on LL-37 parts currently in the registry. The prefix and suffix are compatible with RFC 23 (silver fusion) to allow for protein fusions and this part contains an ???atg??? to allow for immediate protein synthesis. Furthermore this LL-37 derivative does not have any stop codons which allows for composite BioBrick Related parts Previously this part has been used by Slovenia iGEM team 2009 (http://parts.igem.org/wiki/index.php?title=Part:BBa_K245114) This part (BBa_K245114) lacks a start codon. Previously this part has been used Trieste iGEM team 2012 http://parts.igem.org/Part:BBa_K875009?title=Part:BBa_K875009 This part, BBa_K875009, has a BioBrick prefix and suffix of RFC 10 so cannot be used for protein fusions, such as in cases of secretion and protein purification. Also has a stop codon which limits what can be added to the C-terminal end (purification tag/secretion tag).
Antimicrobial peptides (AMPs) are stable peptides that have extensive ability to kill or inhibit the growth of bacteria. They play a role in defense mechanism for all organisms, ranging from prokaryotes to humans, to against predators. Some of them act against specific strain of microbes. Signiferin is a kinds of antimicrobial peptide extracted from the skin mucus of Crinia signifera, respectively. It demonstrated effectiveness in killing MRSA. And we had change it's sequence into E.coli codon usage. We treat a signal peptide in front of signiferin. This signal peptide comes from S.lividas and has ability to let E.coli secrect signiferin to LB culture medium.
We had change signiferin sequence into E.coli codon usage. In order to get the antimicrobial peptide easily we treat signal peptide in front of the AMP.
We synthesis this sequence from IDT (Integrated device technology) company.
Sufactin generator and biofilm repressor for B. subtilis
Ioana Sandu, Gael Chambonnier, Alex Mizgier, Marion Traouan
This part can be used to induce surfactin production (an antimicrobial lipopeptide) and to repress the biofilm formation in B. subtilis strains.
The part has 3 genes (sfp, abrB and lacI) under the control of the xylose inducible Pxyl promoter.
The part was constructed from the assembly of 3 biobricks: BBa_K802006,BBa_K802007 and BBa_K802008.
The part is an coding sequence for an anti-microbial peptides linked to a mussel-foot protein-linked to superfolder GFP for localization. The mussel foot protein will anneal to surfaces as a wet glue and superfolder GFP will allow for florescence imaging and localization. In order to use this part you can produce it in a TAG recoded organism simultaneously expressing a Tyrosine supressor or L-DOPA orthogonal translational system. In order to purify you can use the 2X Strep tag and strep column and later cleave with enterokinase to remove the sequence supressing LL-37 antimicrobial action.
There are linker units between individual components of DNA and restriction sites built for modularity.
This part codes for the Magainin 1 star peptide, a novel peptide design by the University of Melbourne 2014 iGEM team.
Because the protein coding region, Magainin 1, is likely cytotoxic, the protein is expressed using a SUMO fusion partner.
The protein coding region of the peptide was designed de novo by the University of Melbourne 2014 iGEM team. It is based off of the naturally occurring Magainin 1 antimicrobial peptide. The promoter, ribosome binding site, and terminator are standard iGEM parts.
Artilysin Art-175 fused at N-terminal with flagellin 26-47 peptide segment
Artilysins are an exciting class of enzyme-based antibacterials. Their name is derived from "artificial endolysin" and they exploit the lytic power of bacteriophage-encoded endolyins. Endolysins are peptidoglycan hydrolases produced at the end of the lytic cycle that pass through the cytoplasmic membrane, degrade the peptidoglycan layer and cause the osmotic lysis of the infected bacterial cell, thus liberating the progeny. Purified endolysins have been used to kill Gram-positive pathogens. Gram-negative bacteria, however, have a protective outer membrane containing lipopolysaccharide (LPS) that serves as a barrier against the peptidoglycan hydolytic activity of endolysins from the outside. To overcome this problem, selected polycationic or amphipathic peptides that locally destabilize the LPS layer have been covalently fused to endolysins. Biers et al. fused the sheep myeloid antimicrobial peptide (SMAP-29) to the N terminus of the endolysin KZ144 to create Artilysin 175. Art-175 has been shown to be a highly potent antibacterial that acts in minutes to kill virtually all Pseudomonas aeruginosa strains. [1] BBa_K1659001 is a composite of Art-175 with the 26-47 segment of Salmonella typhimurium flagellin combined with a hexahistidine tag. [2] Fla 26-47 is capable of mediating flagellar export of recombinant proteins and, combined with a hexahistidine tag, facilitates their secreted expression and easy purification. [1] Briers, Y., Walmagh, M., Grymonprez, B., Biebl, M., Pirnay, J. P., Defraine, V., ??? Lavigne, R. (2014). Art-175 is a highly efficient antibacterial against multidrug-resistant strains and persisters of Pseudomonas aeruginosa. Antimicrobial Agents and Chemotherapy, 58(7), 3774???3784. http://doi.org/10.1128/AAC.02668-14 [2] Balb??s, P., & Lorence, A. (2004). Recombinant Gene Expression. Reviews and Protocols, 267, 506. http://doi.org/10.1007/978-1-61779-433-9
Export signal combined with hexahistidine tag
Art-175 amino acid sequence: Briers, Y., Walmagh, M., Grymonprez, B., Biebl, M., Pirnay, J. P., Defraine, V., ??? Lavigne, R. (2014). Art-175 is a highly efficient antibacterial against multidrug-resistant strains and persisters of Pseudomonas aeruginosa. Antimicrobial Agents and Chemotherapy, 58(7), 3774???3784. http://doi.org/10.1128/AAC.02668-14 Amino acid sequence of Fla 26-47: Balb??s, P., & Lorence, A. (2004). Recombinant Gene Expression. Reviews and Protocols, 267, 506. http://doi.org/10.1007/978-1-61779-433-9
This part is inserted in BB-NIC-II-HisN vector (BBa_K245005) and can be assembled with other parts according to assembly standard 37.
Pveg + RBS + Antifungal EcAMP + Double terminator
Melittin is an antimicrobial peptide found in the venom of Apis mellifera. It works by aggregating in the membrane of a bacteria cell and forming a pore, ultimately lysing the cell. An original sequence for melittin in the registry can be found in the BBa_K1382000 BioBrick; however, this sequence was taken directly from Apis Mellifera. In this biobrick, the sequence for melittin has been optimized for E. coli.
The sequence was designed to use the codons that are preferred in E. coli.
The original sequence is taken from Apis mellifera; however, this sequence has been optimized for E. coli.
Spheniscin-2 antimicrobial peptide from the king penguin (Aptenodytes patagonicus)
(Chassis E.coli, carrier plasmid pSB1C3, part destined for use in Bacillus subtilis) Length: 1006 bp Background: AF-A is a part designed to be combined with AF-B to form an operon of 5 antifungal peptides. The interest of using several antifungals is to have a broad spectrum of fungi target, because each antifungal has its own characteristics.The peptides were designed to be expressed and secreted by Bacillus subtilis. This operon was also initially designed to be expressed under the control of the pNagA or pNagP promoter (BBa_K1937003 and BBa_K1937005 respectively). In spite of several cloning strategies, the AF-B construction was not obtained, likely due to the toxicity of one of the peptides for Bacillus subtilis. This BioBrick is a part developed by the Toulouse 2016 iGEM team (http://2016.igem.org/Team:Toulouse_France)
This part encodes two genes encoding antifungal peptides: a cutted Metchnikowin and the D4E1. We created it with the end of the Metchnikowin gene because we found out that only this part of the gene had the antifungal property(http://www.ebi.ac.uk/interpro/entry/IPR012513). The beginning of the gene translates for a propeptide. This gene comes from drosophila and translates a proline-rich peptide whose expression is immune-inducible. The peptide is active against fungi.The D4E1 gene is synthetic and translates a peptide that complexes with cholesterol, current in non-germinated conidia. This synthetic peptide is more resistant than the natural peptide CecroprineA because proteases from fungi have difficulties to degrade the synthetic one (http://www.nrcresearchpress.com/doi/abs/10.1139/w98-032?journalCode=cjm#.V_pRgJOLT6Y). So that each antifungal can be secreted out of the cell, it was necessary to add a sequence called AmyE that enables the transport of molecules outside of the cell. As AmyE is used twice in this Biobrick, we modified the AmyE sequences to avoid homologous recombination while keeping the same protein sequence. The NheI restriction site was added after the RBS to allow promoter swapping with biobrick BAAXXXX or BAAXXXX. SacII and SalI restriction site were added to built the full 5 peptides operon.
This gene comes from drosophila and translates a proline-rich peptide whose expression is immune-inducible. The peptide is active against fungi.The D4E1 gene is synthetic and translates a peptide that complexes with cholesterol, current in non-germinated conidia. This synthetic peptide is more resistant than the natural peptide CecroprineA because proteases from fungi have difficulties to degrade the synthetic one (http://www.nrcresearchpress.com/doi/abs/10.1139/w98-032?journalCode=cjm#.V_pRgJOLT6Y).
pBad/araC-RBS B0034-Ag43 thanatin 4mer as alpha domain-double terminator
Canibalism toxin of B. subtilis with modification parts SdpA and SdpB.
Several restriction enzyme sites had to be changed in the coding sequence in order to full fill the BioBrick standard criteria.
EcnA and EcnB from Escherichia coli str. K-12 substr. DH10B
T7 + RBS + EDA + GSG + NdeI + EcnA + TEV + EcnB + 6xHist tag + BamHI + Terminator. Coding device for stabilizing protein EDA linked to EcnA/EcnB through a GSG linker.
Use in conjunction with strain-expressing T7 polymerase.
The human peptide LL-37 is a cationic antimicrobial peptide with activity against Gram-negative and Gram-positive bacteria. It has been shown to protect against Helicobacter pylori infection and its related gastritis, besides of inhibiting the growth of other pathogenic bacteria in the gastrointestinal tract such as Salmonella typhimurium and Escherichia coli.
spaS Subtilin (antimicrobial peptide, Freiburg standard)
Subtilin Lanthionine-containing peptide antibiotic (lantibiotic) active on Gram-positive bacteria. The bactericidal activity of lantibiotics is based on depolarization of energized bacterial cytoplasmic membranes, initiated by the formation of aqueous transmembrane pores.
A new LL 37 bacteria toxin optimized for E. coli expression. The toxin disrupt the membrane from outside and is armless from inside.
This biobrick codes for the promoter pT7, Magainin II peptide with a His 6 tag.
An anti-microbial peptide in use in horseshoe crabs,
Warning: the part is very small which lowers BioBrick standard assembly yields drastically.
Tachypleus Tridentitus, synthesized de novo.
pBad/araC-RBS B0034-Ag43 thanatin 2mer as alpha domain-double terminator
Grammistin-Pp1 antimicrobial peptide from the Clown Grouper (Pogonoperca punctata)
LL-37 antimicrobial peptide generator under the Arabinose promoter.
In the absence of arabinose, AraC protein represses the structural gene by binding to araI1 and araO2 and the DNA forms a loop.
Artilysin Art-175, an antibacterial fusion protein
pSB1C3: 2XStrep_Flagtag_LL-37_Mussel Foot Protein_sfGFP
The part is an coding sequence for an anti-microbial peptides linked to a mussel-foot protein-linked to superfolder GFP for localization. The mussel foot protein will anneal to surfaces as a wet glue and the antimicrobial domain is designed to interact with microbial membranes and interfere with membrane stability. In order to use this part you can produce it in a TAG recoded organism simultaneously expressing a Tyrosine supressor or L-DOPA orthogonal translational system. In order to purify you can use the 2X Strep tag and strep column and later cleave with enterokinase to remove the sequence supressing LL-37 antimicrobial action.
We capped the LL-37 Domain which is otherwise toxic to cells with a removable strep tag that can be used for column chromatography. Additionally there are linker units between individual components
codes for protegrin-1 (anti-microbial peptide)
This is the coding region for protegrin-1.
This sequence was derived from protegrin-1 found in porcine leukocytes. The website where we found the sequence is:
Pveg-spoVG + EcAMP-1 + Double terminator
WAM-1 antimicrobial peptide from the tammar wallaby (Macropus eugenii)
An antimicrobial peptide from the Cathelicidin family of peptides, isolated from the Tammar Wallaby (Macropus eugenii). ATG(Met) added at start of coding region.
Microcin S, a Class II microcin produced by probiotic E. coli G3/10
Microcins are antibacterial peptides. Microcin S was isolated from Escherichia coli G3/10, a component of the probiotic drug Symbioflor 2. Here it was shown to be responsible for E. coli G3/10's ability to suppress adherence of enteropathogenic E. coli E2348/69. [1] The Microcin S operon is about 4.7 kb in size and is comprised of four genes. This part is the sequencing encoding the microcin itself (mcsS). [1] Zsch??ttig, A., Zimmermann, K., Blom, J., Goesmann, A., P??hlmann, C., & Gunzer, F. (2012). Identification and characterization of microcin S, a new antibacterial peptide produced by probiotic Escherichia coli G3/10. PLoS ONE, 7(3), 1???9. http://doi.org/10.1371/journal.pone.0033351
Latherin is a surfactant protein. Its normal biological function is temperature regulation and is believed to function by enhancing evaporation from the pelt. Due to its ability to bind to hydrophobic surfaces, the protein is being investigated for its possible ability to aid in breaking up biofilms. Caution - suspected allergen
Illegal restriction sites - site directed mutagenesis
Originally isolated from horse sweat. Received from researchers