Team:TecMonterrey/030

These are our team's favorite parts

BBa_K1166001

The Fumarate-Nitrate Reductase protein or FNR helps hypoxic promoters to have a more sensitive hypoxic response, it works as an activator and repressor for oxygen-dependant genes. Activity is proven by comparing the expression of GFP (BBa_E0040) by promoters with and without the simultaneous expression of the FNR protein (BAL38390.1).

BBa_K1166002

We assembled this device to save time and ligation reactions for future teams wanting to use the E.coli secretion system. This secretion device includes all the necessary elements (linker-HlyA secretion signal peptide and the expression cassette for HlyB and HlyD) for the secretion of expression cassettes cloned in the 5' terminal of the device.

This secretion device is usable by Silver standard fusion in its 5' end for proteins with no stop codon. Codes for both the proteins HlyB and HlyD (native from uropathogenic strains of E. coli), which, along with the constitutively expressed TolC protein, form a secretion channel. This secretion system recognizes proteins tagged with the C-terminal secretion signal peptide HlyA, and secretes them to the exterior of the bacteria.

Source Parts:

• AraC/pBAD: Bba_I0500
• HlyA: Bba_K554002
• HlyB; Bba_K554007
• HlyD: Bba_K554008

Parts we helped to characterize.

pArs_mRFP (BBa_K1106003)

A promoter sensitive to the presence of the arsenite ion. It codes for mRFP to produce a red fluorescence when in presence of arsenite, working as a biosensor. This part is from Team: Buenos_Aires and was characterized in our lab as a collaboration between TecMonterrey and Buenos_Aires.

New parts we created

Hip-1 (BBa_K1166000)

Composed of the HIP-1 promoter with the FNR expression cassette upstream for a stronger hypoxic regulation, this promoter will start translation in coding sequences 3' downstream in a hypoxic environment.

His-TAT (BBa_K1166003)

A biobrick that includes the TAT signal peptide that can be fused to any protein for an easy internalization into mammal cells. It includes a 6x His-tag for easy purification.

TAT is a cell penetrating peptide (CPP) derived from the transactivator of transcription from HIV and has been used to internalize a range of large and small molecules from peptides, DNA and antibodies, to liposomes.

TRAIL (BBa_K1166004)

Contains the T7 expression cassette for the soluble part (aminoacids 98-281) of TRAIL with an N-terminal histidine tag (x6) to enhance its purification. It is known to cause apoptosis by binding a membrane receptor.

TAT-Apoptin (BBa_K1166005)

Codes for the T7 expression cassette for TAT-Apoptin composed of a N-terminal TAT translocation peptide, followed by a linker (GGGGS), a histidine tag (6x) and by the same linker fused to Apoptin. Apoptin is a protein from the Chicken Anemia Virus known to cause apoptosis in human cancer cell lines only using a pathway independent from P53 by a phosphorilation in the Threonine 108.

Submitted Parts

Registry Number	Type	Short Description	Length
BBa_K1166000	Regulator	FNR regulator under native promoter controlling HIP-1 hypoxic promoter	1234 bp
BBa_K1166001	Regulator	Codes for FNR hypoxic regulator under its native promoter	1168 bp
BBa_K1166002	Device	Silver fusion-ready device for Silver fusion-ready device for secretion using Haemolysin system	5287 bp
BBa_K1166003	Signaling	HIS-TAT peptide for protein internalization	114 bp
BBa_K1166004	Generator	Codes for TRAIL protein under T7 promoter	781 bp
BBa_K1166005	Generator	Codes for TAT-APOPTIN protein under T7 promoter	676 bp