Team:NYMU-Taipei/Project/safe

Introduction

Our Bee. coli expresses some antimicrobial peptides such as defensin and abaecin to fight against Nosema cerenae. There is probability that Bee. coli contaminates the natural environment and cause death to other species. Therefore, light-induced lysis system was created to ensure our Bee. coli only lives inside of the bee.

Background

Blue Light Sensing Device

We chose to use [http://parts.igem.org/Part:BBa_K592016 K592016] as our light sensing device. [http://parts.igem.org/Part:BBa_K592016 K592016] consists two parts: YF1 and FixJ. YF1 IS a blue-light sensor protein. It works in conjunction with its response regulator, FixJ. When exposed to blue-light, they can activate [http://parts.igem.org/Part:BBa_K592006 K592006], the blue-light sensing promoter.

Lysis Device

The lysis device is composed of promoter [http://parts.igem.org/Part:BBa_K592006 K592006], the blue-light sensing promoter, and the lysis protein [http://parts.igem.org/Part:BBa_K896999 K896999], which is a lethal 91 amino acid membrane protein that induces lysis in E. coli.

Circuit design and Experimental Method

Circuit Device

[http://parts.igem.org/Part:BBa_K592016 K592016] is cloned after constitutive promoter [http://parts.igem.org/Part:BBa_J23102 J23102], so the proteins YF1 and FixJ were continuing produced. When exposed to blue light, the inactive YF1 and FixJ will be change to their active form and induce the downstream gene of promoter [http://parts.igem.org/Part:BBa_K592006 K592006]. In our circuit, the lysis protein [http://parts.igem.org/Part:BBa_K896999 K896999] will be produced and kill our Bee. coli which escaped from the midgut of a bee.

Circuit Device

Experimental Method

First, we substitute lysis protein [http://parts.igem.org/Part:BBa_K896999 K896999] with green fluorescent protein [http://parts.igem.org/Part:BBa_E0040 E0040]. By this way, we can test the efficiency of the circuit.

Testing Circuit

Next, by comparing the number of colonies of the plate that is exposed to light and the plate that is blocked from light after 16 hours of in incubator, we can characterize the function of our device.

Left: Light is block by aluminum foil.
Right: Plate is exposed to light.