Streptomyces clavuligerus is the producer of clavulanic acid; aβ-lactamaseinhibitor that is used in combination with the antibiotic amoxicillin. Genome sequencing has established that S. clavuligerus contains a linear chromosome and four linear plasmids: pSCL1, pSCL2, pSCL3 and pSCL4. Although pSCL4 carries 20 % of the S. clavuligerus coding sequences, none are thought to be essential with the exception of tap and tpgthat encode terminal proteins necessary for priming of the lagging strand at the telomeres. In order to confirm plasmid essentiality and the genomic architecture of S. clavuligerus, we first corroborated the available genome sequences by physically mapping the genome using Pulsed-field Gel Electrophoresis, this confirmed the presence of the replicons pSCL2, pSCL3 and pSCL4 with sizes of 150, 450 and 1800 kilobases respectively. In addition, bioinformatics and physical analyses of the S. clavuligerus genome allowed the identification of similar non-archetypal telomeres in the chromosome and pSCL4 at one end of each replicon. Despite this, the other telomere remains unidentified, which suggests there is a dynamic chromosome-plasmid relationship in S. clavuligerus. Furthermore, in order to study the essentiality of pSCL4 we first introduced a copy of tap-tpg onto the chromosome prior to plasmid curing and/or tap-tpg deletion. Consequently, targeted genome sequence and further physical analyses, especially of the telomeres, will permit us understand the complex dynamic relationship between the megaplasmid and the chromosome in this important industrial microorganism.


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