Team:Dundee/Basic Part
Basic Parts
Basic Parts
The sub units of our constructs
BBa_K1962000 ( Colicin Ia )
This sequence codes for the full length Colicin Ia bacteriocin (anti-bacterial toxin). Colicin Ia is a protein that is used by species of E. coli in order to kill closely related species of E. coli in certain circumstances e.g. competition. The producer cell normally contains a protein which confers immunity to this toxin, which in this case is (BBa_K1962001 ). This protein is believed to form a complex with the cytotoxic domain of the bacteriocin rendering it inactive. When this bacteriocin is secreted the immunity protein dissociates from the complex leaving the cytotoxic domain active resulting in the pore forming activity of this bacteriocin returning and having effect on the target cells. The pore forming activity of this bacteriocin results in the depolarisation of the bacterial inner membrane resulting in the collapse of the proton motive force.
BBa_K1962001 (Colicin Ia-Im)
This Biobrick codes for the Immunity Protein for Colicin Ia (BBa_K1962000 ). Colicin Ia is a channel-forming bacteriocin that depolarizes the cytoplasmic inner membrane of target bacteria, leading to dissipation of cellular energy. This Immunity Protein is tightly linked to its specific Colicin bacteriocin domain to protect the colicinogenic cell from the cytotoxic activity of the colicin.
BBa_K1962002 (Truncated Colicin Ia Lacking Bacteriocin Active Domain)
Colicin Ia is a bacteriocin secreted by E. coli. Colicins typically have 3 domains, a translocation domain, a receptor binding domain, and a cytotoxic domain. If you are looking for a full-length Colicin Ia Biobrick it is located here BBa_K1962000 . This part encodes a truncated version of Colicin Ia that lacks the C-terminal bacteriocin domain. The part has an engineered multiple cloning site at the 3' end, preceding the RFC[10] suffix and regulation double stop codons, containing the following in-frame restriction sites: BamHI, KpnI, SalI, BglII and NheI. The presence of this multiple cloning site will allow for different toxic domains to be fused at the C-terminus of the truncated colicin and thereby generating a synthetic colicin in a rapid and facile manner.
BBa_K1962003 (Colicin Ia::Ssp2 Chimera)
This part contains an translational fusion protein between the truncated bacteriocin-free version of Colicin Ia (BBa_K1962002 ) and an antibacterial toxin (peptidoglycan hydrolase) called ssp2 from Serratia marcescens . Ssp2 is an antibacterial effector normally secreted by the Type VI secretion system (T6SS) in Serratia marcescens and it has peptidoglycan endopeptidase activity, degrading peptidoglycan in the periplasm of the target cell where it cleaves between y-D-glutamic acid and L-mesodiaminopimelic acid in peptidoglycan.
BBa_K1962004 ( Immunity Protein Im-E3 )
This Biobrick encodes for the Immunity protein specific for Colicin E3 . This Immunity Protein inhibits the 16S RNA hydrolyzing activity of Colicin E3 by binding with high affinity to the C-terminal catalytic domain of E3 . The sequence was obtained from Uniprot and then codon optimised for E. coli K-12 and synthesised by IDT as a gBlock gene fragment.
BBa_K1962005 (Immunity Protein Im-E9 )
This Biobrick codes for the Immunity Protein that is specific for Colicin E9 . This protein is able to protect a host bacterial cell against attack by Colicin E9 since it binds tightly and specifically to the DNase domain of E9 and inhibits its bactericidal activity.
BBa_K1962006 (Truncated Colicin E9 Lacking Bacteriocin Active
Colicins are anti-bacterial proteins produced by some strains of E. Coli that typically have three domains: a translocation domain; a receptor binding domain; and a cytotoxic domain. This biobrick encodes a truncated version of Colicin E9 which lacks the C-terminal bacterocin domain, which is this case is a DNase. The part has an engineered multiple cloning site at the 3' end, preceding the RFC[10] suffix and regulation double stop codons, containing the following in-frame restriction sites: BamHI, KpnI, SalI, BglII and NheI. The presence of this multiple cloning site will allow for different toxic domains, or other polypeptides, to be fused at the C-terminus of the truncated colicin and thereby generating a synthetic colicin or novel fusion protein in a rapid and facile manner.
BBa_K1962007 ( Colicin E9::Ssp1 Chimera )
This part contains an translational fusion protein between the truncated DNase-domain-lacking version of Colicin E9 (BBa_K1962006 ) and an antibacterial toxin (peptidoglycan hydrolase) called Ssp1 from Serratia marcescens. Ssp1 is an antibacterial effector normally secreted by the Type VI secretion system (T6SS) by Serratia marcescens and it has peptidoglycan endopeptidase activity.
BBa_K1962008 (Colicin E9::Ssp2 Chimera)
This part contains an translational fusion protein between the truncated DNase-domain-lacking version of Colicin E9 (BBa_K1962006 ) and an antibacterial toxin (peptidoglycan hydrolase) called Ssp2 from Serratia marcescens. Ssp2 is an antibacterial effector normally secreted by the Type VI secretion system (T6SS) by Serratia marcescens and it has peptidoglycan endopeptidase activity.
BBa_K1962009 (Transcriptional Activator RamA )
RamA is a transcriptional activator that has been used in conjunction with BBa_K1962010 which is a bile salt sensing device, to induce GFP expression by binding the RamA binding site present on the acrRA promoter.
