Geobacter Investigating a potential mechanism of Mercury Uptake in Geobacter sulfurreducens.

Geobacter

Investigating a potential mechanism of Mercury Uptake in Geobacter sulfurreducens

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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