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

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

  2. And an oxygen sensitive reporter system

  3. Motivation

  4. Microbial Fuel Cells Electricity Produced Bacteria Incoming ½ O 2 Incoming O 2 wastewater ` Outgoing H 2 O Outgoing CO 2 Anode Cathode Semi-permeable membrane

  5. Genetic design NirB RBS LOV Terminator

  6. 4Fe-4S O 2 Ribosome mRNA } FNR FNR Dimerization FNR FNR Translation Transcription RNA How does our openon works? Polymerase FNR= Ferredoxin-NADP+ reductase

  7. The LOV Domain Optimized LOV for Wild type LOV gene from Shewanella oneidensis Bacillus subtilis Removal Cysteine amino acid codon Adding TAA stop codon Added Alanine codon

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

  9. Methods Design Synthesis

  10. Transformations Transformation pSB1C3/RFP pSB1C3/LOV

  11. Electrophoresis pSB1C3/NirB pSB1C3/LOV 4 Kb 2 Kb

  12. 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 **************************************************

  13. Human Practices Waste Water CO2 Anode Cathode

  14. Where do we go from here? ● Retry NirB Plasmid formation ● Finish 3A assembly with LOV and NirB ● Utilizing the microbial fuel cell with Schwenella

  16. Acknowledgements

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