A Synthetic Biology Approach to Heavy Metal Detection

Eric Yang and Qiang ShiEmporia State University

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

Goals of the Model Design

• Simultaneous detection of mercuric and arsenic contamination

• Accumulating and cleaning mercury and arsenic.

Ars Operon

Mercury Operon

Cloning modules

Transporting Module

• GlpF, a aquaglycerol porin from E. coli• Citrate coupled transporters, CitM from

Bacillus subtilis• Promoter + MerR + MerP + MerT

Cloning modules

Detecting Module• Arsenic promoter from E. coli, pArsR + RBS +

GFP• Mercury-Inducible Promoter from E. coli,

pMerR + RBS + RFP

Cloning modules

Sequestering Module• Metallothioneins• pArsR + RBS + GFP + pArsR + RBS + fMT

(arsenic form)• pMerR + RBS + RFP + pMerR + RBS + MerR +

MerP + MerT + fMT (mercuric form)

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

Topological Sort

Let’s Make a Deal

Sudoku

Acknowledgement

• All organizers• All participants

Thank you!