Assessing phenotypes of Thioclava electrotrophica

By Alexandra Marquardt, Biology Advisor: Annette Rowe Presentation ID: PM_D25 Abstract: One of the main challenges of understanding microbial function in environmental systems is the lack of genetic techniques that allow us to assess phenotypes. This has been especially true in chemolithoautotrophs which are often challenging to grow in the lab. To aim this, we have been developing techniques for transferring genetic information into the facultative chemolithoautotroph, Thioclava electrotrophica ElOx9. Having developed a media compatible for growth with both E. coli and T. electrotrophic, we have successfully developed a conjugation protocol that has allowed us to transform T. electrotrophica with both a fluorescent marker and introduce the minMar transposon. In this work we will present the mating efficacy for this process. In order, assess future compatibility with more additional genetic techniques like CRISPR CAS-9 mediated genome editing, we have also been investigating the potential for electroporation, stemming from recent work published for Shewanella. Our current work suggested that Thioclava are capable of withstanding the electroporation conditions, which hold promise for the potential of this approach to yield genetic transformation. Our future work will entail using these techniques to construct a library of gene deletion mutants using transposon mutagenesis that can be used to assess the genes involved in chemolithoautotrophic growth of T. electrotrophica on elementals sulfur and electron uptake from electrodes, to further study the potential genes involved in these processes.