Developing a genetic toolbox for Aeromonas salmonicida subsp. pectinolytica to analyze function and interplay of its native SMC-complexes Smc-ScpAB and MukBEF

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SMC protein complexes condense chromosomes by binding and translocating along the DNA and thereby are an essential part of compaction of the DNA of a cell. The both most important SMC-complexes in bacteria, Smc-ScpAB and MukBEF, compact the DNA in a different manner: While Smc-ScpAB aligns both chromosome arms, MukBEF condenses each chromosome arm separately. In contrast to all other bacteria, which contain either Smc-ScpAB or MukBEF, species of Aeromonas harbor both SMC-complexes. This co-appearance allows the analysis of differences in function and roles of Smc-ScpAB and MukBEF regarding chromosomal segregation and conformation, as well as their coordination in the same system. To allow such an analysis in Aeromonas, genetic tools like plasmids, inducible promoters and techniques for genetic manipulation of Aeromonas are needed. Hence, the aim of this project is to establish a genetic toolbox for Aeromonas salmonicida subsp. pectinolytica and to analyze the function and interplay of Smc-ScpAB and MukBEF.