1. A Synthetic Biology Approach to Heavy Metal Detection Eric Yang and Qiang Shi Emporia State University

2. Heavy Metal Poisoning Arsenic, mercury, lead, antimone, cadmium, etc. Contamination of water and soil Oxidative stress to cells --- damage to DNA and proteins Disrupt function in vital organs and glands Example: Mercury Effects – adrenal dysfunction, kidney damage, allergy, alopecia, anorexia, anxiety, birth defects, blushing, brain damage, cataracts, cerebral palsy, poor coordination / jerky movements, deafness, depression, dermatitis, discouragement, dizziness, drowsiness, eczema, emotional disturbances, excess saliva, fatigue, gum bleeding and soreness, headaches (band type), hearing loss, hyperactivity, hypothyroidism, forgetfulness, immune dysfunction, insomnia, irritability, joint pain,, loss of self-control, memory loss, mental retardation, metallic taste, migraines, nervousness, nerve fiber degeneration, numbness, pain in limbs, rashes, retinitis, schizophrenia, shyness, speech disorders, suicidal tendencies, tingling, tremors (eyelids, lips, tongue, fingers, extremities), vision loss, weakness

3. Goals of the Model Design Simultaneous detection of mercuric and arsenic contamination Accumulating and cleaning mercury and arsenic.

4. Ars Operon

5. Mercury Operon

6. Cloning modules Transporting Module GlpF, a aquaglycerol porin from E. coli Citrate coupled transporters, CitM from Bacillus subtilis Promoter + MerR + MerP + MerT

7. Cloning modules Detecting Module Arsenic promoter from E. coli, pArsR + RBS + GFP Mercury-Inducible Promoter from E. coli, pMerR + RBS + RFP

8. Cloning modules Sequestering Module Metallothioneins pArsR + RBS + GFP + pArsR + RBS + fMT (arsenic form) pMerR + RBS + RFP + pMerR + RBS + MerR + MerP + MerT + fMT (mercuric form)

9. Cloning modules Buoyancy Module gas vesicle protein gene cluster from Bacillus megaterium, GVP pArsR + RBS + GFP + pArsR + RBS + fMT (arsenic form) + RBS + GVP pMerR + RBS + RFP + pMerR + RBS + MerR + MerP + MerT + fMT (mercuric form) + RBS + GVP

10. Topological Sort

11. Let’s Make a Deal

12. Sudoku

13. Acknowledgement All organizers All participants

14. Thank you!