Thioclava electrotropha ElOx9 Tn-Seq

To explore the genetic basis of oxidative EET, we utilized genomic analyses and transposon insertion mutagenesis screens (Tn-seq) in the metabolically flexible lithotrophic Alphaproteobacterium Thioclava electrotropha ElOx9. The polished genome of this strain is 4.3 MB, and consists of 4,139 predicted ORFs, 54 contain heme binding motifs, and 33 of those 54 are predicted to localize to the cell envelope or have unknown localizations. To begin to understand the genetic basis of oxidative EET in ElOx9, we constructed a transposon mutant library comprised of over 91,000 individual mutants encompassing more than 69,000 unique TA dinucleotide insertion sites in semi-rich media and subjected the library to heterotrophic growth on minimal media with acetate and autotrophic oxidative EET conditions on indium tin oxide coated glass electrodes poised at -278 mV vs. SHE, with electrochemistry pre-growth with minimal media and acetate, and open circuit conditions serving as controls.