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Geobacter Investigating a potential mechanism Mercury Uptake in Geobacter sulfurreduce
7

Geobacter

Feb 24, 2016

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Investigating a potential mechanism of Mercury Uptake in Geobacter sulfurreducens. Geobacter. Geobacter sulfurreducens PCA chromosome. 1463922. 1468286. Heavy Metal Binding Domain containing protein. ABC Transporter Permease. CxxC motif. Graphic from NCBI database. - PowerPoint PPT Presentation
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Page 1: Geobacter

Geobacter

Investigating a potential mechanism of Mercury Uptake in Geobacter sulfurreducens

Page 2: Geobacter

1463922 1468286

Geobacter sulfurreducens PCA chromosome

Heavy Metal Binding Domain containing protein

CxxC motif

ABC Transporter Permease

Graphic from NCBI database

Page 3: Geobacter

Sequence AlignmentsHMA domainGMTCxxC

Signal peptidePeriplasmic location

Page 4: Geobacter

GSU1338 and merP comparison

Next Step:

Page 5: Geobacter

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

Page 6: Geobacter

Ladder Total 3 4 5 6 7 8 9 10 Induced8/9

10 kda

Page 7: Geobacter

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