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Geobacter
Investigating a potential mechanism of Mercury Uptake in Geobacter sulfurreducens
1463922 1468286
Geobacter sulfurreducens PCA chromosome
Heavy Metal Binding Domain containing protein
CxxC motif
ABC Transporter Permease
Graphic from NCBI database
Sequence AlignmentsHMA domainGMTCxxC
Signal peptidePeriplasmic location
GSU1338 and merP comparison
Next Step:
Experimental Approach
• PCR • DNA extraction from gel• Enzyme Digestion & Ligation• Transformation• Harvest and sequence• Incubate cells until OD600 .4-.6 • Induce with IPTG to final concentration .4mM• Run HisLink Nickel Column
Ladder Total 3 4 5 6 7 8 9 10 Induced8/9
10 kda
Next step: competition experiments
• Binding assays– Use merP as + control– Have – control– Titrate protein into assay with Hg bound to something that we
understand i.e. cysteine
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