Eco-friendly E. coli

Introduction

Our Bee. coli can express antimicrobial peptides such as defensin and abaecin to fight against Nosema cerenae. However, it is also possible that our Bee. coli can contaminate the natural environment and cause death to other species. Therefore, a 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 K592016 as our light sensing device. 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 K592006, the blue-light sensing promoter.

Lysis Device

The lysis device is composed of promoter K592006, the blue-light sensing promoter, and the lysis protein K896999, which is a lethal 91 amino acid membrane protein that induces lysis in E. coli.

Circuit design and Experimental Method

Circuit Design

K592016 is cloned after constitutive promoter 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 K592006. In our circuit, the lysis protein 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 K896999 with green fluorescent protein 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 incubation, we can characterize the functions of our device.

Left: Light is block by aluminum foil.

Right: Plate is exposed to light.