Cellulose binding Domain from C. Fimi with const. Promotor and GFP
Cellulose binding Domain from Cellulomonas Fimi with const. Promotor J61101 and Reporter GFP
Upon recognition of a small molecule (e.g. nutrients or toxins), chemoreceptors transduce a signal through a set of Che proteins. This group of methylesterases and phosphatases regulates the rotation of the flagella. At the end of the chain of Che proteins is CheY. In phosphorylated state, CheY binds the flagella and causes the cells to tumble. The phosphatase CheZ dephosphorylates CheY, inducing dissociation of CheY from the flagella enabling the cells to swim. Cells lacking the CheZ protein are unable to swim and will tumble excessively and incessantly.
1 bp in BBa_K629003 was adapted to create an original restriciton site in the plasmid. CheZ was fused to GFP (BBa_K082003) and its LVA tag was codon-optimized to avoid introducing artificial DNA repetitions.
DNA sequences originate from the E. Coli genome and previous biobrick sequences. The ribosome binding site was chosen from the Anderson RBS family.
CBD of C. cellulovorans cellulose binding protein gene with const. Promoter (J23100+J61101) and GFP
Cellulose binding Domain of Clostridium cellulovorans [http://www.ncbi.nlm.nih.gov/nuccore/M73817 cellulose binding protein gene (cbp A)] with a constitutive promoter and RBS (<partinfo>J61101</partinfo>) and a fused-GFP protein with His-tag.
Was made to switch from a T7-promoter to a constitutive promoter
Cellulose binding domain form a cloning-vector. <partinfo>J61101</partinfo> and [http://partsregistry.org/Part:BBa_I13522 GFP] from the partsregistry.
Autoinducer synthetase for acyl-homoserine lactones (AHL): LuxI synthesizes AHL (3OC6HSL). Two molecules of AHL form a complex with two molecules of the LuxR protein. This complex then binds to a palyndromic site on the lux promoter, increasing its transcription rate. LuxI, LuxR and the right lux promoter are all derived from V. fischeri. In absence of AHL, the right lux promotor only gives weak constitutive expression of downstream genes. The LuxI gene is fused to a His-tag facilitating gene expression measurements.
Biobrick BBa_C0061 still contained a Barcode sequence which was deleted. Also, a Ribosome Binding Site was added. For our own use, 5 bp were exchanged to create unique restriction sites and to optimize DNA folding.
Sequence is originating from V. fischeri. Here, the sequence from BBa_C0061 was used and optimized.
This part has the arabinose regulated promoter (AraC/pBAD) upstream of a ribosome binding site. This is ideal for inserting a protein to be expressed downstream of this part as it's expression can be regulated by the presence of the concentration of arabinose in the growing environment/media of the cells. ==What this part actually does== This composite part includes the AraC/pBAD promoter and a strong ribosome binding site designed from the Anderson family of RBS. The AraC/pBAD promoter can be used with prokaryotic chassis to regulate expresion of proteins. When the AraC/pBAD promoter is placed upstream of your canditate translational unit (protein to be expressed) it allows control of the trancription of the said protein. This is done via controlling the concentration of arabinose sugar in the environment/solution of bacterial growth. Controlling the transcription will not ensure translation. The ribosome binding site is ideally placed a small distance away from the promoter site to ensure that the ribosome will bind and start translating the protein coding sequence found downstream.
The part was designed with RFC10 assembly from supplied biobricks of the iGEM2009 distribution.
The AraC promoter can be found naturally at 1.52min of the E.coli genome ([http://ecogene.org/geneinfo.php?eg_id=EG10054 EcoGene number EG10054]. The RBS was taken from the designed sequences of RBS by the Anderson family. For more information visit the original part page [http://partsregistry.org/Part:BBa_J61100 here].
Promoter Anderson RBS GFP double terminator
This is an RBS from the Anderson catalogue.
A part used to quantify the AraC-RBS construct <partinfo>K259007</partinfo> created by the BCCS-Bristol iGEM'09 team.
This part was designed with the specified biobricks from the parts registry. AraC occurs naturally in E.coli. GFP occurs in jellyfish species, RBS designed by the Anderson family and Terminator by Resma Shetty.