Team:UPO-Sevilla/Basic Part

BASIC PARTS

BBa_K1973000

Codifies the LapG protein from Pseudomonas putida KT2442, a periplasmic protease that participates in the biofilm dispersion process by cleaving adhesion proteins such as LapA.

BBa_K1973001

nasF is an attenuator of transcription elongation (terminator) from Klebsiella pneumoniae that decreases the basal expression from the Psal promoter. The NasR protein disrupts the terminator structure and allows the expression again, providing an useful tool for tight ON/OFF expression systems.

BBa_K1973002

Codifies the modified XylS2 protein from Pseudomonas putida MT1, which is a transcriptional activator of the Pm promoter in response to salicylate. The modification allows the response to salicylate. See BBa_K1031911 (original version of this part, which is regulated by benzoate and derivatives of this).

BBa_K1973013

Pm1 is one variant of the promoter for the xylS/Pm expression system. It was obtained by site-directed mutagenesis PCR from the Pseudomonas putida plasmid pMPO52 in order to remove illegal restriction sites. The expression of this promoter is activated by XylS or XylS2 in the presence of benzoate and salicylate, respectively.

BBa_K1973014

Pm2 is one variant of the promoter for the xylS/Pm expression system. It was obtained by site-directed mutagenesis PCR from the Pseudomonas putida plasmid pMPO52 in order to remove illegal restriction sites. The expression of this promoter is activated by XylS or XylS2 in the presence of benzoate and salicylate, respectively.

BBa_K1973015

Pm3 is one variant of the promoter for the xylS/Pm expression system. It was obtained by site-directed mutagenesis PCR from the Pseudomonas putida plasmid pMPO52 in order to remove illegal restriction sites. The expression of this promoter is activated by XylS or XylS2 in the presence of benzoate and salicylate, respectively.

BBa_K1973016

Codifies the PleD* protein from Caulobacter crescentus, which is a diguanylate cyclase that induces the biofilm formation by increasing c-di-GMP levels. Contains a constitutive mutation in the original sequence (see BBa_K2012002) that improves the enzymatic activity by knocking out the regulation system.

BBa_K1973026

Codifies the GlpF protein from Pseudomonas aeruginosa, which is a glycerol channel that increases the glycerol intake for bioremediation purposes.

BBa_K1973030

Codifies the Epi protein from Pseudomonas mosselii, which participates in the propionate metabolism by catalyzing the conversion between R-Methylmalonyl-CoA and S-Methylmalonyl-CoA.

BBa_K1973033

Contain the module nahR-Psal. nahR (Pseudomonas putida, see BBa_K1031610) is expressed under the constitutive Pr promoter. It encodes a regulatory protein that activates the Psal promoter (see BBa_J61051) in the presence of salicylate.

BBa_K1973036

lapC encodes the LapC protein, which is an essential protein for the secretion of adhesion proteins (such as LapA) that are involved in the biofilm formation process. The creation of a lapC- mutant avoids the secretion of these molecules, hence blocking biofilm formation.

BBa_K1973037

NasR (Chai and Stewart, 1998) is RNA-binding protein that inhibits transcription termination dependent on the nasF terminator. nasF (see BBa_K1973001) is an attenuator of transcription elongation (terminator) from Klebsiella pneumoniae that decreases the basal expression from the Psal promoter.

BBa_K1973034

yhjH encodes the YhjH protein, a c-diGMP (cyclic diguanilate monophosphate) phosphodiesterase. c-diGMP regulates both the formation and dispersion of the biofilm, depending on the quantity of it that is present inside the cell. High levels of c-diGMP will increase the biofilm formation, and low levels of it will induce its dispersion. To manipulate this, we will make use of a phosphodiesterase, that will destroy the c-diGMP, and a diguanilate cyclase, that will produce it, YhjH and PleD* respectively.