%0 Journal Article %A Bocioaga, Daniela %A El Gheriany, Iman A. %A Lion, Leonard W. %A Ghiorse, William C. %A Shuler, Michael L. %A Hay, Anthony G. %T Development of a genetic system for a model manganese-oxidizing proteobacterium, Leptothrix discophora SS1 %D 2014 %J Microbiology, %V 160 %N 11 %P 2396-2405 %@ 1465-2080 %R https://doi.org/10.1099/mic.0.079459-0 %I Microbiology Society, %X Understanding the molecular underpinnings of manganese oxidation in Leptothrix discophora SS1 has been hampered by the lack of a genetic system. In this report, we describe the development of a genetic system for L. discophora SS1. The antibiotic sensitivity was characterized, and a procedure for transformation with exogenous DNA via conjugation was developed and optimized, resulting in a maximum transfer frequency of 5.2×10−1 and a typical transfer frequency of the order of 1×10−3 transconjugants per donor. Genetic manipulation of L. discophora SS1 was demonstrated by disrupting pyrF via chromosomal integration with a plasmid containing a R6Kγ origin of replication through homologous recombination. This resulted in resistance to 5-fluoroorotidine, which was abolished by complementation with an ectopically expressed copy of pyrF cloned into pBBR1MCS. This system is expected to be amenable to a systematic genetic analysis of L. discophora SS1, including those genes responsible for manganese oxidation. %U https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.079459-0