Development of a genetic system for a model manganese-oxidizing proteobacterium, Leptothrix discophora SS1

作者：Bocioaga Daniela; El Gheriany Iman A; Lion Leonard W; Ghiorse William C; Shuler Michael L; Hay Anthony G^*

来源：Microbiology-SGM, 2014, 160(Pt_11): 2396-2405.

DOI：10.1099/mic.0.079459-0

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 5.2x10(-1) and a typical transfer frequency of the order of 1 x10(-3) transconjugants per donor. Genetic manipulation of L. discophora SS1 was demonstrated by disrupting pyrFvia chromosomal integration with a plasmid containing a R6K gamma 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.

• 出版日期2014-11