Functional characterization of two putative mobile elements of the 'Mycoplasma mycoides cluster'
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[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] Horizontal gene transfer has played a major role in bacterial evolution, including in the genus Mycoplasma, which is comprised of some of the simplest and smallest free-living organisms. This dissertation focuses on the functional characteristics of two recently characterized putative mobile elements of the ruminant-infecting Mycoplasma mycoides cluster: a 3.4-kb cryptic plasmid, pMyBK1, of Mycoplasma yeatsii, and an approximately 24-kb member of the mycoplasmal family of Integrative, Conjugative Elements (ICE), ICEC, of Mycoplasma capricolum subsp. capricolum strain Kid. Vectors were constructed using pMyBK1 as a backbone and replication of vector derivatives in M. yeatsii identified the ORF required for survival of the plasmid. Showing the utility of the vector system, heterologous proteins were expressed in M. yeatsii from pMyBK1 derivatives. To determine which ORF of ICEC acts as an integrase, mini-ICEC derivatives were constructed. The right-hand terminal ORF, MCAP_0571, was shown to be the ORF required to integrate the mini-element into Mycoplasma capricolum subsp. capricolum strain GM262G.
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