Isolation and characterization of a linear DNA plasmid from Streptomyces clavuligerus

Keen, Cheryl L.; Mendelovitz, Simona; Cohen, Gerald; Aharonowitz, Yair; Roy, Kanak

doi:10.1007/bf00322461

Cited by 40 publications

(27 citation statements)

References 19 publications

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“…1 The two terminal fragments of pSCL1, the 0.9-kb segment from the left and the 2.8-kb segment from the right, were isolated by SalI digestion. The 5' termini of pSCL1 are blocked in vivo by terminal proteins, and even after treatment with proteinases, one or more amino acid residues remain (23). Since the amino acid-DNA linkage at the 5' termini prevents ligation of the terminal fragments, direct cloning of these two fragments was not thought possible.…”

Section: Resultsmentioning

confidence: 99%

“…During the past year, portions of the terminal sequences of SCP1 have also been published (28). In a previous study, we reported the isolation and characterization of a 12-kb linear plasmid (pSCL1) in S. clavuligerus, a ,-lactam antibiotic producer (23). It was demonstrated in that study that, like most other linear plasmids, pSCL1 has proteins covalently linked to the termini of its DNA and contains approximately 900-bp terminal repeats.…”

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confidence: 99%

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Complete nucleotide sequence of a linear plasmid from Streptomyces clavuligerus and characterization of its RNA transcripts

Roy

1993

J Bacteriol

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The complete nucleotide sequence of a small linear plasmid (pSCL1) from Streptomyces clavuligerus has been determined. This plasmid is 11,696 bp in length, has a 72% G+C content, and has =900-bp inverted terminal repeat sequences. A comparison of the inverted terminal repeats of pSCL1 with those of a linear plasmid from S. rochei shows that the two terminal sequences have a high degree of similarity (=70%) (19). One of the most notable features of this genus is that many members are capable of producing antibiotics. Because of their consequent medical and industrial importance, much attention has been focused on the genetics of this and related genera, particularly with regard to the organization and regulation of genes involved in antibiotic biosynthesis (18,56). Most of the genes directing the synthesis of antibiotics in streptomycetes appear to be chromosomal. However, pulse-field gel electrophoresis experiments have shown that a number of antibiotic-producing Streptomyces spp. harbor giant linear plasmids (27). Moreover, plasmid SCP1 in Streptomyces coelicolor A3(2), which has been demonstrated by genetic methods (29) to encode methylenomycin biosynthesis and resistance genes, is a 350-kb giant linear plasmid (26). Less convincing is the evidence suggesting that a 520-kb linear plasmid (pKSL) may be required for the production of lasalocid and echinomycin by S. lasaliensis (26,27) and that a 17-kb linear plasmid (pSLA2) may be involved in the production of the lankacidin group of antibiotics by S. rochei (15).Among the prokaryotes, linear plasmids have been found largely in Streptomyces spp. and other actinomycetes (e.g., 22), although they have also been found in a wide variety of eukaryotic organisms (12). The linear plasmids in eukaryotes are almost all double-stranded DNA molecules, with proteins covalently linked to their 5' termini. These have been completely sequenced, including two mitochondrial plasmids in Zea mays, S1 and S2 (45), and three nonmitochondrial plasmids in fungi, pGKL-1 and pGKL-2 in Kluyveromyces lactis (59, 67) and pClK-1 in Claviceps purpurea (43). Almost all linear plasmids sequenced to date have long inverted terminal repeats and include a putative DNA polymerase gene.Recently, a different type of linear plasmid, having covalently closed ends at the termini of its DNA, was reported for Escherichia coli harboring plasmid prophage N15 (63), for the bacterium Borrelia burgdorferi (2), as well as for the fungus Rhizoctonia solani (37).Although eukaryotic linear plasmids have been extensively studied, much less is known of linear plasmids in Streptomyces spp. Until recently, the only sequence information reported for Streptomyces linear plasmids was the 614-bp terminally repeated sequence of pSLA2 from S. rochei (17) and the mmr region of SCP1 (42). During the past year, portions of the terminal sequences of SCP1 have also been published (28). In a previous study, we reported the isolation and characterization of a 12-kb linear plasmid (pSCL1) in S. clavuligerus, a ,-lactam antibiot...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Complete nucleotide sequence of a linear plasmid from Streptomyces clavuligerus and characterization of its RNA transcripts

Roy

1993

J Bacteriol

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“…In S . clavuligerus, plasmid pSCL (1 2 kb) (Keen et al, 1988) was found, and in S. rirnosus, pSRM (42 kb) (Chardon-Loriaux et al, 1986). Outside this genus, in Borrelia hermsii and B. burgdorferi linear plasmids of 28 kb and 49 kb, respectively, were detected (Plasterk et al, 1985 ;Barbour & Garon, 1987).…”

Section: Discussionmentioning

confidence: 99%

Hydrogen autotrophy of Nocardia opaca strains is encoded by linear megaplasmids

Kalkus

Reh

Schlegel

1990

Journal of General Microbiology

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Several linear megaplasmids were detected in the facultatively lithoautotrophic Gram-positive bacterium Nocardia opaca The wild-type strain M R l l contains, in addition to the cccDNA plasmids pHG31-a and pHG31-b, the linear plasmids pHG201(270 kb), pHG202 (400 kb) and pHG203 (420 kb). The wild-type strain MR22 contains, in addition to the cccDNA plasmid pHG33, the linear plasmids pHG204 (180 kb), pHG205 (280 kb) and pHG206 (510 kb). After preparation of DNA from cells embedded in agarose, the linear plasmids were demonstrated by pulsed-field electrophoresis. By means of DNA probes for genes of soluble hydrogenase and ribulose-bisphosphate carboxylase, the conjugative plasmids pHG201 and pHG205 were shown to be the carriers of the genetic information for these enzymes. A restriction map of pHG201 for the enzymes AsnI, SpeI, XbaI is presented.

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“…Unlike most other bacterial chromosomes, the~8 Mb chromosomes of Streptomyces are linear and Streptomyces species often harbour linear as well as circular plasmids (Hayakawa et al, 1979;. The known linear plasmids range in size from 12 to 1800 kb (Keen et al, 1988;Medema et al, 2010). Their 'telomeres' contain inverted repeat sequences from 44 to 180 kb long Pandza et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Three functional replication origins of the linear and artificially circularized plasmid SCP1 of Streptomyces coelicolor

et al. 2013

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Previous reports showed that the large linear plasmid SCP1 of Streptomyces coelicolor A3(2) contains a 5.4 kb centrally located replication locus. We report here that SCP1 actually contains three internal replication loci. Subcloning of the 5.4 kb sequence identified a 3.2 kb minimal locus (rep1A/repB/iteron) that determined propagation in Streptomyces lividans. The two newly identified replication genes, rep2A and rep3A, resembled the rep gene of Streptomyces circular plasmid pZL12. Transcription start points of the three replication genes were determined. The three replication loci could independently determine propagation in linear mode in S. lividans. Individual and sequential deletions of the rep1A and rep3A genes were successful. The SCP1-derived linear plasmids with deletions of the rep1A and/or rep3A genes still propagated in similar copy numbers, were inherited largely stable and were transferred efficiently by conjugation in S. coelicolor. Interestingly, SCP1 can be artificially circularized to yield a 280 kb circular plasmid, circular SCP-1 (C-SCP1), which contains the three replication loci. Strikingly, the copy numbers, inheritance and transfer of C-SCP1 resembled that of the linear SCP1 plasmids. Transcripts of the rep1A, rep2A and rep3A genes in linear or artificially circularized SCP1 were detected at all the time points of strain growth.

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Isolation and characterization of a linear DNA plasmid from Streptomyces clavuligerus

Cited by 40 publications

References 19 publications

Complete nucleotide sequence of a linear plasmid from Streptomyces clavuligerus and characterization of its RNA transcripts

Complete nucleotide sequence of a linear plasmid from Streptomyces clavuligerus and characterization of its RNA transcripts

Hydrogen autotrophy of Nocardia opaca strains is encoded by linear megaplasmids

Three functional replication origins of the linear and artificially circularized plasmid SCP1 of Streptomyces coelicolor

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