In B. subtilis the extracellular matrix is composed of two major components, an EPS and the protein TasA, which polymerizes into amyloid-like fibers.
SlrR is an important transcriptional regulator in Bacillus subtilis. SlrR represses the sigma(D)-dependent flagellar genes and activates the eps and yqxM operons. Therefore it is has a controlling factor in biofilm formation. By overexpressing SlrR, with the use of the promoter Pveg and a strong RBS, a robust biofilm can be formed.
This gene was ordered via IDT, hence it was not biobrick compatible.
Ordered via IDT based on the genomic sequence from Bacillus subtilis 168
Lysostaphin generator for B. subtilis
Lysostaphin is a bacterial biocide isolated from Staphylococcus simulans.
Used a strong RBS in order to produce high levels of expression of Lysostaphin.
This is a composite part including the constitutive B. subtilis Pveg promotor and lysostaphin coding sequence.
This part is based on BBa_K729004, modified by deleting the stop codons. It's also fusion with HlyA secretional tag for extracelullar expression
This part makes the Nuclease secreted extracelullar
It was modified from BBa_K729004, BBa_K554002
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.
This part encodes E coli protein Ag43, with an additional 6xHis tag for easy purification and quantification. Ag43 expression is known to cause E coli cells to adhere together, and the 6xHis tag was added to allow for convenient study of variation of E coli aggregation with respect to Ag43 expression levels.
Ag43 is an outer membrane E coli protein, so it has a N-terminal 52 amino acid long signal peptide sequence, which is cleaved after Ag43 pro-protein is transported into the periplasmic space. Since strong dependence has been demonstrated on the signal peptide cleavage on the signal peptide-protein junction, the 6xHis tag should be added after Ala53, the residue immediately after the signal peptide sequence.
This comes from Gibson assembly of fragments from a previous biobrick, BBa_K1352000 by Team:Aberdeen_Scotland, 2014.
OmpR234 protein, with higher effect on Curli promoter
This part is coding for a mutated protein OmpR. This protein will be phosphorilated by the complex Envz. The phosphorilated protein is an activator of the Curli promoter
This is a fusion protein with Ag43, a protein from E coli tagged with a 6xHis tag on the N-terminal and sfGFP tagged on C-terminal of the pro-protein. Ag43 causes cell-cell adhesion in E coli, and the 6xHis and sfGFP tags are present to allow convenient study dependence of Ag43 mediated aggregation on Ag43 expression levels, which can be determined from the sfGFP fluoresence and/or quantification of 6xHis tag. The 6xHis tag is included to allow protein concentration estimation in case the sfGFP fails to fold properly.
The location for the 6xHis tag is the same as that for BBa_K2168000(Ag43 tagged with 6xHis on N-terminal under pBAD/araC promoter) and uses the same design principles. For independent folding, a flexible GGGGSGGGGS linker was added between Ag43 and sfGFP, and the linker was choosen to be not very hydrophilic or hydrophobic, since Ag43 lies very close to lipid membranes.
This part was made from Gibson assemblies from fragments of two previous parts - BBa_K1352000 and BBa_I746908, which code for Ag43 and sfGFP respectively.
Dispersin B with mutation at nucleotide 582 (T to C)
he Dispersin gene codes for an enzyme which catalyzes the hydrolysis of the extracellular matrix produced by Gram negative bacteria.With strong constitutive promoter,together with ribosomal binding site and signal sequence yebF, make dispersin work more accurately.
To fit igem part standard, the circuit is constructed in backbone pSB1C3, chloramphenicol with EcoR1 and Xba1 in front of the part, and Spe1, Pst1 behind the part.
The promoter, J23100, comes from the 2014 distributed igem kit. The signal sequence, yebF, encoding signal protein exporting recombinant protein extracellular, is from E.coli K-12. By designing primer, we obtain it together with its naturally occurring ribosomal binding site using PCR and extraction.The dispersin is from Lyon INSA igem team. Finally, the terminator B0015 is from 2014 igem distribution.
Dispersin generator for B. subtilis
The part contains the DspB gene coding for an enzyme which catalyzes the hydrolysis of the extracellular matrix produced by Gram-negative bacteria.
Used a strong RBS in order to produce high levels of protein.
Dispersin is produced by Aggregatibacter actinomycetemcomitans.
A enzyme that is know to degrade biofilms.