Download - Geobacter Investigating a potential mechanism of Mercury Uptake in Geobacter sulfurreducens
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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
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Sequence AlignmentsHMA domainGMTCxxC
Signal peptidePeriplasmic location
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GSU1338 and merP comparison
Next Step:
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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
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Ladder Total 3 4 5 6 7 8 9 10 Induced8/9
10 kda
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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