Team:JiangnanU China/Parts
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Basic Parts
BBa_K3137000 : rsmH
250bp constitutive promoter
RsmH can detect the intensity of the fluorescence of gfp.
RsmH can detect the intensity of the fluorescence of gfp.
BBa_K3137002 : PputA
250bp inducible promoter
PputA is an inducible promoter. When E.coli is infected by a phage, both report and respond gene circuits are induced to express at the same time. When a phage infects Escherichia coli into the latent period (about 5 min), PputA will induce the bacteria to express anti-protein and display green fluorescence.
PputA is an inducible promoter. When E.coli is infected by a phage, both report and respond gene circuits are induced to express at the same time. When a phage infects Escherichia coli into the latent period (about 5 min), PputA will induce the bacteria to express anti-protein and display green fluorescence.
BBa_K3137003 : PglcF
250bp inducible promoter
PglcF is an inducible promoter that works when phages infect bacterial for around 20 min. If the anti-proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the anti-proteins cannot kill phages, the phages will continue to infect. When phages infect bacteria into the burst period (about 20 min), the inducible promoter PglcF will induce expression of the red fluorescence protein gene mCherry and the downstream toxic protein gene protegrin-1, which will make the bacteria display red fluorescence and lyse cells before the assembly of phages.
PglcF is an inducible promoter that works when phages infect bacterial for around 20 min. If the anti-proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the anti-proteins cannot kill phages, the phages will continue to infect. When phages infect bacteria into the burst period (about 20 min), the inducible promoter PglcF will induce expression of the red fluorescence protein gene mCherry and the downstream toxic protein gene protegrin-1, which will make the bacteria display red fluorescence and lyse cells before the assembly of phages.
BBa_K3137004
mCherry 708bp report gene
The mCherry can express red fluorescence protein and it is used as a report gene in the process of reporting the number of cells.
The mCherry can express red fluorescence protein and it is used as a report gene in the process of reporting the number of cells.
BBa_K3137005
gntR 996bp
The Gluconate repressor gntR, is a transcription factor that negatively regulates the operon involved in the catabolism of d-gluconate via the Entner-Doudoroff pathway and represses genes involved in the different systems related to d-gluconate uptake: gluconate I and gluconate II.
The Gluconate repressor gntR, is a transcription factor that negatively regulates the operon involved in the catabolism of d-gluconate via the Entner-Doudoroff pathway and represses genes involved in the different systems related to d-gluconate uptake: gluconate I and gluconate II.
BBa_K3137006
abpAB 3803bp anti-protein
Gene abpAB is from E.coli genome which express anti-proteins that are resistant to T2,T4,T7 and λ phages. We obtained abpA and abpB by PCR from the genome of E.coli BL21 and constructed recombinant plasmid that connected abpA and abpB at the same time. Then we used IPTG to induce the expression of anti-protein and verified the resistant function of the protein.
Gene abpAB is from E.coli genome which express anti-proteins that are resistant to T2,T4,T7 and λ phages. We obtained abpA and abpB by PCR from the genome of E.coli BL21 and constructed recombinant plasmid that connected abpA and abpB at the same time. Then we used IPTG to induce the expression of anti-protein and verified the resistant function of the protein.
BBa_K3137007
rzpD 462bp
Overexpression of rzpD causes an abnormal biofilm architecture.
Overexpression of rzpD causes an abnormal biofilm architecture.
BBa_K3137008
yhjH 768bp
YhjH protein contains a EAL domain to catalyze c-di-GMP into GMP, so as to regulate the levels of c-di-GMP.
YhjH protein contains a EAL domain to catalyze c-di-GMP into GMP, so as to regulate the levels of c-di-GMP.
BBa_K3137009
nuoE 501bp
NuoE is part of the soluble fragment of NADH dehydrogenase I, which represents the electron input part of the enzyme.
NuoE is part of the soluble fragment of NADH dehydrogenase I, which represents the electron input part of the enzyme.
BBa_K3137010
T7 terminator 48bp terminator
T7 terminator is used to quantify the level of expression in E. coli.
T7 terminator is used to quantify the level of expression in E. coli.
BBa_K3137011
gfp 717bp reporter
The gfp can express green fluorescent protein, which can be used as a report gene in the process to report the number of cells.
The gfp can express green fluorescent protein, which can be used as a report gene in the process to report the number of cells.
BBa_K3137011
codes forprotegrin-1 (anti-microbial peptide) 60bp
antibacterial peptides
Protegrin-1 coding region. Protegrin-1 is an 18-residue beta-sheet peptide isolated from porcine leukocytes with antimicrobial activity against a broad range of microorganisms. It performs its effect by pore membrane disruption and possibly also by effects such as activation of membrane-damaging proteases and has anti-microbial activity against E. coli.
Protegrin-1 coding region. Protegrin-1 is an 18-residue beta-sheet peptide isolated from porcine leukocytes with antimicrobial activity against a broad range of microorganisms. It performs its effect by pore membrane disruption and possibly also by effects such as activation of membrane-damaging proteases and has anti-microbial activity against E. coli.
Composite Part
BBa_K3137013
report circuit 1666bp
The report gene circuit consists of two periods of phage infection. In incubation period (about 5 min),
there is an inducible promoter PputA and a green fluorescent protein gene gfp. In outbreak
period (about
20 min), there is an inducible promoter PglcF, and a red fluorescent protein gene mCherry.
When E. coli is infected by a phage, both report and respond gene circuits are induced to express
at the
same time. When a phage infects E. coli into the incubation period (about 5 min), the inducible
promoter
PputA will induce the bacteria to express the anti-proteins and the green fluorescent protein
gene
gfp.
If the anti-protein successfully resists phage infection, the inducible promoter PglcF will
not
induce the expression of downstream gene. If the anti-protein fails to kill phages, it means that
phages will continue to infect bacteria. And when it comes to the burst period (around 20 min) of phage
infection, the inducible promoter PglcF will activate the red fluorescent protein gene
mCherry and the
downstream toxic protein gene so that the bacteria will display red fluoresce and lyse cells before the
assembly of phages.
BBa_K3137014
respond circuit 5097bp
The respond gene circuit is composed of an inducible promoter PputA and anti-phage protein genes of the
latent period (about 5 min), and an inducible promoter PglcF and a toxic protein gene protegrin-1 of
the burst period (about 20 min).
When E. coli is infected by a phage, both report and respond gene circuits are induced to express at the
same time. When a phage infects E. coli into the latent period (about 5 min), the inducible promoter
PputA will induce the bacteria to express the resistant protein abpAB and gntR, and the green
fluorescent protein gene gfp.
If the resistant proteins successfully resist phage infection, the inducible promoter PglcF will not
induce the expression of downstream gene. If the anti-proteins fail to kill phages, it means that
phages will continue to infect bacteria, when it comes to the burst period (around 20 min) of phage
infection, the inducible promoter PglcF will activate the red fluorescent protein gene mCherry and
downstream toxic protein gene protegrin-1 so that the bacteria will display red fluoresce
and lyse cells before the assembly of phages.
