1986
DOI: 10.1128/mcb.6.1.80
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Replicating plasmids in Schizosaccharomyces pombe: improvement of symmetric segregation by a new genetic element.

Abstract: We characterized a number of widely used yeast-Escherichia coli shuttle vectors in the fission yeast Schizosaccharomyces pombe. The 2,um vectors pDB248 and YEp13 showed high frequency of transformation, intermediate mitotic and low meiotic stability, and a low copy number in S. pombe, analogous to their behavior in [ciro] strains of Saccharomyces cerevisiae. The S. cerevisiae integration vectors pLEU2 and pURA3 transformed S. pombe at very low frequencies but, surprisingly, in a nonintegrative fashion. Instea… Show more

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Cited by 133 publications
(71 citation statements)
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“…In S. pombe, the region sufficient for autonomous replication cannot be delimited shorter than 1-2 kb, and no essential sequences have been deduced (Johnston and Baker, 1987;Maundrell et al, 1988). It has been shown that several plasmids that do not transform S. pombe at a high frequency can autonomously replicate as a highly polymeric form in transformants formed at a low frequency (Heyer et al, 1986). These observations suggest that requirement for specific sequences in S.…”
Section: Discussionmentioning
confidence: 89%
“…In S. pombe, the region sufficient for autonomous replication cannot be delimited shorter than 1-2 kb, and no essential sequences have been deduced (Johnston and Baker, 1987;Maundrell et al, 1988). It has been shown that several plasmids that do not transform S. pombe at a high frequency can autonomously replicate as a highly polymeric form in transformants formed at a low frequency (Heyer et al, 1986). These observations suggest that requirement for specific sequences in S.…”
Section: Discussionmentioning
confidence: 89%
“…Cultures used for DNA extraction were grown overnight at 25 uC in 10 ml of YPGL on a rotary shaker. The cells were harvested by centrifugation, washed once with distilled water, resuspended in 0?2 citrate/phosphate buffer (pH 5?6) containing 4 mg Zymolyase ml 21 and incubated at room temperature for 1-1?5 h. Genomic DNA was then isolated and purified as described for Schizosaccharomyces pombe (Heyer et al, 1986).…”
Section: Methodsmentioning
confidence: 99%
“…S. pombe homothallic strain GP1596 (h 90 rec11-111 ade6-M26 ura4-294 leu1-32) was transformed to Leu þ with plasmid pYL84, containing the ade6-469 gene which can recombine with the chromosome to produce Ade þ recombinants. The resulting ᮊ 1997 Blackwell Science Ltd, Molecular Microbiology, 23, 869-878 The S. pombe stb element stabilizes the plasmid in mitotic and meiotic cells, and the S. pombe ars element aids mitotic propagation (Heyer et al, 1986). The plasmid contains the S. pombe ura4 þ gene for selection in S. pombe (Grimm et al, 1988), and the pUC19 amp and ori elements for selection and propagation in E. coli (Yanisch-Perron et al, 1985).…”
Section: Screen For Rec11 Clone Isolationmentioning
confidence: 99%
“…As pFL20, the parent of the initial clone pYFL119, is multicopy (Heyer et al, 1986), the clone pYFL119 might contain either the rec11 gene or a multicopy suppressor. To distinguish between these possibilities, disruptions were made by inserting a Saccharomyces cerevisiae LEU2 DNA fragment into the EcoRI site of pYFL104 and into the SwaI site of pYFL105 (Fig.…”
Section: Gene Disruptionmentioning
confidence: 99%