Rapid small-scale plasmid isolation by several methods from filamentous cyanobacteria

Lambert, Grant R.; Carr, N. G.

doi:10.1007/bf00413524

Cited by 33 publications

(23 citation statements)

References 16 publications

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“…In addition to chromosomal DNA, plasmids are found in many cyanobacteria (Asato & Ginoza 1973, Simon 1978, Lau et al 1980, Reaston et al 1980, Lambert & Carr 1982, but the role of the plasmids in the cells are not known. Some studies have been suggested that the extrachromosomal genes in cyanobacteria could be involved in conferring resistance to heavy metals and antibiotics (Olafson et al 1979, Singh & Pandey 1982, and in the production of gas vacuoles (Walsby 1977).…”

Section: Cell Inclusionsmentioning

confidence: 99%

Cell composition and metal tolerance in cyanobacteria

Fiore

Trevors

1994

Biometals

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Section: Cell Inclusionsmentioning

confidence: 99%

Cell composition and metal tolerance in cyanobacteria

Fiore

Trevors

1994

Biometals

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“…coli was grown in L broth, supplemented as appropriate with none, one, or two of the following: 25 jig of chloramphenicol (Cm) per ml, 10-25 pg of streptomycin (Sm) per ml, 50 ,ug of kanamycin (Km) per ml, 50 ,g of ampicillin (Ap) per ml. Plasmids were isolated from cyanobacteria by established methods (5,21,22) or minor variations thereof, and from E. coli strains HB101 and Gm48 (dam-dcm-) essentially as described (23). Plasmids were digested with restriction enzymes from New England BioLabs and Bethesda Research Laboratories in buffers recommended by the suppliers, or slight variations thereof.…”

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confidence: 99%

Construction of shuttle vectors capable of conjugative transfer from Escherichia coli to nitrogen-fixing filamentous cyanobacteria.

Wölk¹,

Vonshak²,

P³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

287

180

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Wild-type cyanobacteria of the genus Anabaena are capable of oxygenic photosynthesis, differentiation of cells called heterocysts at semiregular intervals along the cyanobacterial filaments, and aerobic nitrogen fixation by the heterocysts. To foster analysis of the physiological processes characteristic of these cyanobacteria, we have constructed a family of shuttle vectors capable of replication and selection in Escherichia coli and, in unaltered form, in several strains of Anabaena. Highly efficient conjugative transfer of these vectors from E. coli to Anabaena is dependent upon the presence of broad host-range plasmid RP-4 and of helper plasmids. The shuttle vectors contain portions of plasmid pBR322 required for replication and mobilization, with sites for Anabaena restriction enzymes deleted; cyanobacterial replicon pDUl, which lacks such sites; and determinants for resistance to chloramphenicol, streptomycin, neomycin, and erythromycin.Many filamentous cyanobacteria fix dinitrogen aerobically within specialized cells called heterocysts that differentiate at semiregular intervals along the filaments. All cyanobacteria are capable of oxygenic photosynthesis. Genetic methods usable for study of nitrogen fixation, differentiation, pattern formation, and photosynthesis in these organisms have long been sought.Several unicellular cyanobacteria can be transformed by DNA in the growth medium (1-3). Shuttle vectors, plasmids able to replicate in Escherichia coli and in an alternative host, have been constructed for two such strains, Anacystis nidulans strain R2 and Agmenellum quadruplicatum ). An Anacystis gene cloned in a shuttle vector in E. coli was returned to the cyanobacterium by transformation (7). To date, no reproducible transformation system is known for filamentous cyanobacteria.Conjugation provides an alternative approach to transfer of cloned DNA. RP-4 and related plasmids can transfer themselves or derivatives of themselves into a wide range of We report the construction of a hybrid between pBR322 and plasmid pDU1 (16) from the filamentous cyanobacterium Nostoc. Because restriction endonucleases present in strains of cyanobacteria (17) apparently reduce retention of DNA transferred into those cyanobacteria (4, 18), the hybrid plasmid was restructured to eliminate sites for restriction enzymes present in several strains of Anabaena. Additional antibiotic-resistance determinants were added, lest an organism be unable to use any one such determinant. The derivatives of the hybrid plasmid proved to be shuttle vectors, capable of RP-4-mediated transfer into several strains of Anabaena and of replication in those strains.MATERIALS AND METHODS Anabaena sp. PCC 7120, Anabaena sp. U. Leningrad strain 458 (PCC 7118), Anabaena sp. U. Tokyo M-131, and Nostoc sp. PCC 7524 were grown with nitrate as described (19). Anacystis nidulans strain R2 was grown in medium BG-11 (20).E. coli was grown in L broth, supplemented as appropriate with none, one, or two of the following: 25 jig of chloramphenicol (Cm) per...

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“…Plasmids were isolated from unicellular cyanobacteria using the method of Felkner and Barnum (1988) and the gentle lysis procedure of Lambert and Carr (1982). Both the methods yielded plasmids in Microcystis aeruginosa PCC7820 and M. aeruginosa M228.…”

Section: Resultsmentioning

confidence: 99%

“…(b) Plasmid isolation by gentle lysis procedure Plasmids were isolated from unicellular and filamentous cyanobacteria using gentle cell lysis procedure of Lambert and Carr (1982).…”

Section: Methodsmentioning

confidence: 99%

Plasmid distribution among unicellular and filamentous toxic cyanobacteria

Bose

Carmichael

1990

J Appl Phycol

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Plasmid content of 5 hepatotoxin and 2 neurotoxin producing cyanobacterial strains were analyzed. Among the hepatotoxin-producing strains, Microcystis aeruginosa PCC7820, M. aeruginosa M228 and M. aeruginosa UV027 were found to carry plasmids, whereas other hepatotoxin and neurotoxin producing strains did not harbor any plasmids. Correlations were sought between toxicity and the presence of plasmids in toxic cyanobacteria as a function of age. Aged cultures of M. aeruginosa PCC7820 exhibited toxicity and harbored plasmids. In other cyanobacterial strains, plasmids were not detected. The data add to and support the current understanding that plasmids are probably not involved in toxin production in cyanobacteria.

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Rapid small-scale plasmid isolation by several methods from filamentous cyanobacteria

Cited by 33 publications

References 16 publications

Cell composition and metal tolerance in cyanobacteria

Cell composition and metal tolerance in cyanobacteria

Construction of shuttle vectors capable of conjugative transfer from Escherichia coli to nitrogen-fixing filamentous cyanobacteria.

Plasmid distribution among unicellular and filamentous toxic cyanobacteria

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