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

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Presentation transcript:

Geobacter Investigating a potential mechanism of Mercury Uptake in Geobacter sulfurreducens

Geobacter sulfurreducens PCA chromosome Heavy Metal Binding Domain containing protein CxxC motif ABC Transporter Permease Graphic from NCBI database

Sequence Alignments HMA domain GMTCxxC Signal peptide Periplasmic location

GSU1338 and merP comparison Over-express in E. coli Determine transformation and induction protocol Purify Isolate protein through Nickel His- Link column Competition Experiments Next Step:

Experimental Approach PCR DNA extraction from gel Enzyme Digestion & Ligation Transformation Harvest and sequence Incubate cells until OD Induce with IPTG to final concentration.4mM Run HisLink Nickel Column

Ladder Total Induced 8/9 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