• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Jul 1, 1992; 89(13): 6129–6133.
PMCID: PMC402135

Reconstruction of a Streptomyces linear replicon from separately cloned DNA fragments: existence of a cryptic origin of circular replication within the linear plasmid.

Abstract

We report here the reconstruction of a functional linear replicon, the 12-kilobase Streptomyces clavuligerus plasmid pSCL, from separate DNA fragments cloned in Escherichia coli on the pUC19 plasmid. Protein-free DNA molecules containing the full-length pSCL sequence, an internally inserted thiostrepton-resistance gene, and adventitious nucleotides external to the pSCL termini were introduced into Streptomyces lividans, where the synthesis and functional attachment of replication proteins occurred and pSCL was established as an extrachromosomal linear replicon. Transformation of S. lividans with uncut supercoilded pUC19/pSCL DNA from E. coli or with a circularized 8-kilobase internal fragment of pSCL yielded circular replicons, indicating the existence of a cryptic origin of circular replication within the linear plasmid. Insertion mutations at sites that prevented the replication of pSCL linear plasmids also interfered with its replication in the circular mode.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Hirochika H, Sakaguchi K. Analysis of linear plasmids isolated from Streptomyces: association of protein with the ends of the plasmid DNA. Plasmid. 1982 Jan;7(1):59–65. [PubMed]
  • Kinashi H, Shimaji M, Sakai A. Giant linear plasmids in Streptomyces which code for antibiotic biosynthesis genes. Nature. 328(6129):454–456. [PubMed]
  • Keen CL, Mendelovitz S, Cohen G, Aharonowitz Y, Roy KL. Isolation and characterization of a linear DNA plasmid from Streptomyces clavuligerus. Mol Gen Genet. 1988 Apr;212(1):172–176. [PubMed]
  • Leblond P, Francou FX, Simonet JM, Decaris B. Pulsed-field gel electrophoresis analysis of the genome of Streptomyces ambofaciens strains. FEMS Microbiol Lett. 1990 Oct;60(1-2):79–88. [PubMed]
  • Solenberg PJ, Baltz RH. Transposition of Tn5096 and other IS493 derivatives in Streptomyces griseofuscus. J Bacteriol. 1991 Feb;173(3):1096–1104. [PMC free article] [PubMed]
  • Sakaguchi K. Invertrons, a class of structurally and functionally related genetic elements that includes linear DNA plasmids, transposable elements, and genomes of adeno-type viruses. Microbiol Rev. 1990 Mar;54(1):66–74. [PMC free article] [PubMed]
  • Stillman BW. The replication of adenovirus DNA with purified proteins. Cell. 1983 Nov;35(1):7–9. [PubMed]
  • Salas M. Initiation of DNA replication by primer proteins: bacteriophage phi 29 and its relatives. Curr Top Microbiol Immunol. 1988;136:71–88. [PubMed]
  • Omer CA, Cohen SN. Plasmid formation in Streptomyces: excision and integration of the SLP1 replicon at a specific chromosomal site. Mol Gen Genet. 1984;196(3):429–438. [PubMed]
  • Hirochika H, Nakamura K, Sakaguchi K. A linear DNA plasmid from Streptomyces rochei with an inverted terminal repetition of 614 base pairs. EMBO J. 1984 Apr;3(4):761–766. [PMC free article] [PubMed]
  • Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. [PMC free article] [PubMed]
  • Thompson CJ, Kieser T, Ward JM, Hopwood DA. Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. Gene. 1982 Nov;20(1):51–62. [PubMed]
  • Zalacain M, González A, Guerrero MC, Mattaliano RJ, Malpartida F, Jiménez A. Nucleotide sequence of the hygromycin B phosphotransferase gene from Streptomyces hygroscopicus. Nucleic Acids Res. 1986 Feb 25;14(4):1565–1581. [PMC free article] [PubMed]
  • Lyra C, Savilahti H, Bamford DH. High-frequency transfer of linear DNA containing 5'-covalently linked terminal proteins: electroporation of bacteriophage PRD1 genome into Escherichia coli. Mol Gen Genet. 1991 Aug;228(1-2):65–69. [PubMed]
  • Gutiérrez J, García JA, Blanco L, Salas M. Cloning and template activity of the origins of replication of phage phi 29 DNA. Gene. 1986;43(1-2):1–11. [PubMed]
  • Hay RT, Stow ND, McDougall IM. Replication of adenovirus mini-chromosomes. J Mol Biol. 1984 Jun 5;175(4):493–510. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...