Antibiotics for the 21st Century

While vital to our quality of life, traditional antibiotics face the serious problems of widespread bacterial resistance and destruction of natural gut flora - problems which call for improved twenty-first century antibiotics. Using synthetic biology tools, we designed, built, and tested two new systems to fight infections by both broad types of bacteria - Gram-positive and Gram-negative. Our first project targets Bacillus anthracis, the Gram-positive pathogen that causes anthrax. We re-engineered an enzyme to remove the pathogen's protective coating, rendering it defenseless against the immune system. In our second project, we re-engineered and transplanted a protein secretion system capable of combating Gram-negative bacteria into E. coli. This system was designed to target Gram-negative pathogens in a modular and controllable fashion. These two systems are the vanguard of a new era of antibiotics using the power of nature harnessed with the tools of synthetic biology.

will include intro image, idea is currently being fleshed out from sketch into image

21st century antibiotics GENERAL STORYLINE FLOW

The bullet points are the key images that need to be generated. They should be very general and easy to understand! The text will basically be explaning the pictures and telling the story.... think of this as a comic book!

gram(+)

summary

• why anthrax is bad
• proteins fighting anthrax

design

• circular permutation
• computational design

build

• How we made mutants
• How we made protiein

Test

• gel/mass spec cp vs D
• activity cp vs D
• catalytic KO of CP vs CP
• CP mutations
• Specificity switch

Future Directions

• Just some bullet points, no figures needed

gram(-)

summary (bugs vs bugs)

• type 6 secretion (spear) done-Matthew Harger
• Toxin/Antitoxin produced in prescence of XXX (poison)

design

• operon in Pseudomonas —> operon in e coli ( fosmid)
  • what parts of this operon are needed? justin has this
• sensor —produces—> toxin/antitoxin (classic tinkercell picture)

build

• exporter —> recombineering to promoter image already made
• sensor/toxin: —> PCR —> HSL biobrick vector made by Matthew Harger

Test

• Exporter: Still under construction
• Sensor/Toxin
  • Toxin is toxic in ecoli: (Rachael hood,2010) paper; Sensor works and produces tse2 as expected ( include image of western blot with annotations)

Antidote still works in e.coli indirect evidence, state lack of mutations, may not need image.;

Future Directions

• Just some quick Bullet Points