Part:BBa_K1373001

nadE with strong promter and strong RBS

nadE gene overexpression

Electricity active cells (EAC) generate electrons by organic substrate metabolism and transfer them through extracellular transport to an electrode surface in Microbial fuel cell (MFCs). Therefore, genetic modifications that increase the amount of electrons in EAC is one of feasibilities to optimize the electricity power output of MFCs.

Recent studies have proved that the intracellular redox state of EAC is one of the most important physiological traits of extracellular electron transfer efficiency. Particularly, the NAD+(H) pool size plays a central role of most metabolic pathways. In this study, we aim that, overexpression of gene nadE which encodes a NAD synthetase and catalyzes the final step in de novo synthesis and salvage pathway of NAD biosynthesis (Fig. 1), may increase the NAD+ level. Therefore the augmented pool size of NAD+(H) result in promotion of NADH (the carrier of electrons) level, leading to high generation of intracellular releasable electrons and better electricity performance of EAC.

In our project, BBa_K1373001 and BBaK1373002 are two new parts for over-expression of nadE (K1373000[1]) gene, based on two existing biobricks BBa_K608002 (strong constitutive promoter with strong RBS) and BBa_K608010 (weak constitutive promoter with Strong RBS), respectively. We confirmed the mRNA and protein expression level of NAD synthetase in corresponding genetically modified strains with BBa_K1373001 or BBa_K1373002 by semi-quantitative RT-PCR and SDS-PAGE assay.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]