And an oxygen sensitive reporter system Motivation Microbial Fuel - - PowerPoint PPT Presentation

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Jun 01, 2023 201 likes •381 views

And an oxygen sensitive reporter system Motivation Microbial Fuel Cells Electricity Produced Bacteria Incoming O 2 Incoming O 2 wastewater ` Outgoing H 2 O Outgoing CO 2 Anode Cathode Semi-permeable membrane Genetic design NirB

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And an oxygen sensitive reporter system

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Motivation

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` Incoming O2 Outgoing CO2 Incoming wastewater

½ O2

Anode Cathode Semi-permeable membrane Electricity Produced

Bacteria

Outgoing H2O

Microbial Fuel Cells

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Genetic design

NirB RBS LOV Terminator

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FNR FNR RNA Polymerase

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Dimerization mRNA Ribosome 4Fe-4S O2 FNR FNR

How does our openon works?

FNR= Ferredoxin-NADP+ reductase Transcription Translation

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The LOV Domain

Removal Cysteine amino acid codon

Added Alanine codon Wild type LOV gene from Bacillus subtilis Optimized LOV for Shewanella oneidensis Adding TAA stop codon

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Sterilizing Work Stations
Personal Safety
Equipment testing
Proper disposal of waste materials
Proper supervision

Safety

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Synthesis Design

Methods

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Transformations

pSB1C3/LOV

Transformation

pSB1C3/RFP

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4 Kb 2 Kb pSB1C3/LOV pSB1C3/NirB

Electrophoresis

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Sequencing

CLUSTAL 2.1 multiple sequence alignment LOV --------------------------------------------GAATTC 14 02-027-08-VF2_D11.ab1 TTCNGATAAAAAAAATCCTTAGCTTTCGCNANNGNNGATTTCTGGAATTC 100 02-027-06-VF2_C11.ab1 TTCAGATAAAAAAAATCCTTAGCTTTCGCNNAGGANGATTTCTGGAATTC 99 ****** LOV GCGGCCGCTTCTAGAGATGGCTTCTTTCCAATCTTTCGGTATCCCAGGTC 64 02-027-08-VF2_D11.ab1 GCGGCCGCTTCTAGAGATGGCTTCTTTCCAATCTTTCGGTATCCCAGGTC 150 02-027-06-VF2_C11.ab1 GCGGCCGCTTCTAGAGATGGCTTCTTTCCAATCTTTCGGTATCCCAGGTC 149 ************************************************** LOV AACTGGAAGTTATCAAAAAAGCTCTGGATCACGTTCGTGTTGGTGTTGTT 114 02-027-08-VF2_D11.ab1 AACTGGAAGTTATCAAAAAAGCTCTGGATCACGTTCGTGTTGGTGTTGTT 200 02-027-06-VF2_C11.ab1 AACTGGAAGTTATCAAAAAAGCTCTGGATCACGTTCGTGTTGGTGTTGTT 199 ************************************************** LOV ATCACTGATCCAGCTCTGGAAGATAACCCAATCGTTTACGTTAACCAAGG 164 02-027-08-VF2_D11.ab1 ATCACTGATCCAGCTCTGGAAGATAACCCAATCGTTTACGTTAACCAAGG 250 02-027-06-VF2_C11.ab1 ATCACTGATCCAGCTCTGGAAGATAACCCAATCGTTTACGTTAACCAAGG 249 ************************************************** LOV TTTCGTTCAAATGACTGGTTACGAAACTGAAGAAATCCTGGGTAAAAACG 214 02-027-08-VF2_D11.ab1 TTTCGTTCAAATGACTGGTTACGAAACTGAAGAAATCCTGGGTAAAAACG 300 02-027-06-VF2_C11.ab1 TTTCGTTCAAATGACTGGTTACGAAACTGAAGAAATCCTGGGTAAAAACG 299 ************************************************** LOV CTCGTTTCCTGCAAGGTAAACACACTGATCCAGCTGAAGTTGATAACATC 264 02-027-08-VF2_D11.ab1 CTCGTTTCCTGCAAGGTAAACACACTGATCCAGCTGAAGTTGATAACATC 350 02-027-06-VF2_C11.ab1 CTCGTTTCCTGCAAGGTAAACACACTGATCCAGCTGAAGTTGATAACATC 349 ************************************************** LOV CGTACTGCTCTGCAAAACAAAGAACCAGTTACTGTTCAAATCCAAAACTA 314 02-027-08-VF2_D11.ab1 CGTACTGCTCTGCAAAACAAAGAACCAGTTACTGTTCAAATCCAAAACTA 400 02-027-06-VF2_C11.ab1 CGTACTGCTCTGCAAAACAAAGAACCAGTTACTGTTCAAATCCAAAACTA 399 ************************************************** LOV CAAAAAAGATGGTACTATGTTCTGGAACGAACTGAACATCGATCCAATGG 364 02-027-08-VF2_D11.ab1 CAAAAAAGATGGTACTATGTTCTGGAACGAACTGAACATCGATCCAATGG 450 02-027-06-VF2_C11.ab1 CAAAAAAGATGGTACTATGTTCTGGAACGAACTGAACATCGATCCAATGG 449 **************************************************

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Human Practices

Waste Water Anode Cathode CO2

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Retry NirB Plasmid formation
Finish 3A assembly with LOV and NirB
Utilizing the microbial fuel cell with

Schwenella

Where do we go from here?

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