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  • Articles  (135)
  • Transformation  (135)
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  • Articles  (135)
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  • Springer  (135)
  • Cambridge University Press
  • Nature Publishing Group
  • Periodicals Archive Online (PAO)
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  • 1
    Electronic Resource
    Electronic Resource
    Transformation ofGaeumannomyces graminis to benomyl resistance (1988)
    Springer
    Current genetics 14 (1988), S. 113-117 
    Details
    ISSN: 1432-0983
    Keywords: Gaeumannomyces graminis ; Take-all disease ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Gaeumannomyces graminis var.graminis andtritici were transformed to benomyl resistance using pBT3, a plasmid encoding fungicide-resistant β-tubulin. Either circular or linear plasmid DNA producedG. graminis var.graminis transformants in which plasmid DNA was integrated into the fungal genome. There was no evidence for autonomous plasmid replication in any of the transformants examined. 4/11 linear DNA transformants had a single plasmid copy, whereas 8/9 circular DNA transformants had multiple copies of the plasmid. Integration of transforming DNA occurred by nonhomologous recombination in all (20/20) of these transformants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00569334
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  • 2
    Electronic Resource
    Electronic Resource
    Heterologous gene expression of the glyphosate resistance marker and its application in yeast transformation (1989)
    Springer
    Current genetics 15 (1989), S. 91-98 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Vector ; Glyphosate resistance ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The E. coli aroA gene was inserted between yeast promoter and terminator sequences in different shuttle expression plasmids and found to confer enhanced EPSP synthase activity as well as resistance to glyphosate toxicity. Subsequently, a transformation system using these newly constructed vectors in yeast was characterized. The efficiency of the glyphosate resistance marker for transformation and selection with plasmid pHR6/20-1 in S. cerevisiae laboratory strain SHY2 was found to be relatively high when compared with selection for LEU2 prototrophy. The fate of the recombinant plasmid pHR6/20-1 in the transformants, the preservation of the aroA E. coli DNA fragment in yeast, mitotic stability, EPSP synthase activity, and growth on glyphosate-containing medium have been investigated. As this plasmid also allows direct selection for glyphosate resistant transformants on rich media, the glyphosate resistance marker was used for transforming both S. cerevisiae laboratory strain SHY2 and brewer's yeast strains S. cerevisiae var. “uvarum” BHS5 and BHS2. In all cases, the vector pHR6/20-1 was maintained as an autonomously replicating plasmid. The resistance marker is, therefore, suitable for transforming genetically unlabeled S. cerevisiae laboratory, wild, and industrial yeast strains.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435454
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  • 3
    Electronic Resource
    Electronic Resource
    Expression of invertase activity in Yarrowia lipolytica and its use as a selective marker (1989)
    Springer
    Current genetics 16 (1989), S. 253-260 
    Details
    ISSN: 1432-0983
    Keywords: Yarrowia lipolytica ; Invertase ; Secretion ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Few selective markers are available for the transformation of the industrial yeast Yarrowia lipolytica, and those that are require the use of specialized hosts (e.g., auxotrophs, antibiotic sensitive). To enable the transformation of any Y. lipolytica strain, we used the property that Y. lipolytica cannot use sucrose as a sole carbon source. We have constructed a gene fusion where the Saccharomyces cerevisiae SUC2 gene is placed under the control of the promoter and signal sequence of the Y. lipolytica XPR2 gene, which encodes an Alkaline Extracellular Protease (AEP). Strains bearing this fusion express invertase activity and grow on sucrose as a carbon source. The activity follows the same regulation as does the alkaline extracellular protease, is secreted into the periplasm and confers a Suc+ phenotype. It was shown that this chimeric gene could be used as a dominant marker for transformation in a one-step procedure.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422111
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  • 4
    Electronic Resource
    Electronic Resource
    Behaviour of recombinant plasmids in Aspergillus nidulans: structure and stability (1986)
    Springer
    Current genetics 10 (1986), S. 767-775 
    Details
    ISSN: 1432-0983
    Keywords: Aspergillus ; Transformation ; Recombinant plasmids ; Autonomously replicating sequences
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A pyrG − Aspergillus strain was transformed with plasmid pDJB-1, derived from pBR325 by insertion of the Neurospora crassa pyr4 gene (orotidine 5′-phosphate carboxylase), giving mitotically unstable transformants. Aspergillus DNA which acted as an “autonomously replicating sequence” (ARS) in yeast was inserted into pDJB-1 and the resulting construct, pDJB12.1, gave mitotically stable transformants when introduced into Aspergillus. Transformants obtained with pDJB-1 and pDJB12.1 gave few pyr − progeny in crosses to a pyrG + strain. Southern hybridisation analysis of pyr + transformants obtained with pDJB-1 revealed restriction fragments expected for integrated plasmid but transformants obtained with pDJB12-1 showed only bands derived from free plasmid. pDJB-1 and derivatives of pDJB12.1 could be recovered from transformants. These derivatives could not be explained by straightforward excision of integrated pDJB12.1 sequences but could result from recombination between plasmid molecules. Hybridisation of undigested transformant DNAs showed that the transforming DNA was present in a high molecular weight form. These results suggest: (1) pDJB12.1 derivatives and possibly pDJB-1 can replicate autonomously in Aspergillus; (2) A. nidulans DNA acting as an ARS in yeast enhances replication and/or segregation of transforming plasmids in Aspergillus; and (3) recombinant plasmids may undergo rearrangements when introduced into Aspergillus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00405100
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  • 5
    Electronic Resource
    Electronic Resource
    Evidence for autonomous replication and stabilization of recombinant plasmids in the transformants of yeast Hansenula polymorpha (1986)
    Springer
    Current genetics 10 (1986), S. 741-747 
    Details
    ISSN: 1432-0983
    Keywords: Transformation ; Recombinant plasmids ; H. polymorpha
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary For the transformation of the yeast Hansenula polymorpha we have constructed a set of hybrid plasmids carrying the LEU2 gene of Saccharomyces cerevisiae as a selective marker and fragments of mitochondrial DNA of Candida utilis and H. polymorpha or chromosomal DNA fragments of H. polymorpha as replicator sequences. The replication properties of chimeric plasmids in the yeast H. polymorpha were investigated. We showed that for plasmids propagated autonomously in this yeast the plasmid monomers could be detected in the transformants only during the immediate time after the transformation event. Further growth under selective conditions led to the selection of polymeric forms of plasmid DNA as it was clearly shown for transformants carrying cosmid pL2 with mtDNA fragment of C. utilis. Such transformants carrying polymerized plasmids showed a remarkably increased stability of the transformed phenotype. Cosmid pL2 was able to shuttle between Escherichia coli, S. cerevisiae and H. polymorpha, whereas plasmids with DNA fragments from H. polymorpha did not transform S. cerevisiae effectively.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00405096
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  • 6
    Electronic Resource
    Electronic Resource
    Identification of a lys2 mutant of Candida maltosa by means of transformation (1987)
    Springer
    Current genetics 11 (1987), S. 385-391 
    Details
    ISSN: 1432-0983
    Keywords: Candida maltosa ; Transformation ; LYS2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated five mutants of Candida maltos, which lack the 2-aminoadipate reductase activity, an enzyme involved in the lysine biosynthesis. By means of complementation analysis using protoplast fusion, the isolated mutants were divided into two complementation groups. Thereof the C. maltosa strain G457 could be transformed by the plasmids pDP12 and pDP13, which contain the L YS2-coding gene of Saccharomyces cerevisiae. On the basis of our presented results obtained by studies on hybridization, stability, and recovery of plasmids from C. maltosa transformants, we suggest that transformation does proceed integratively.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00378181
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  • 7
    Electronic Resource
    Electronic Resource
    High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier (1989)
    Springer
    Current genetics 16 (1989), S. 339-346 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Transformation ; ss carrier DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A method, using LiAc to yield competent cells, is described that increased the efficiency of genetic transformation of intact cells of Saccharomyces cerevisiae to more than 1 × 105 transformants per microgram of vector DNA and to 1.5% transformants per viable cell. The use of single stranded, or heat denaturated double stranded, nucleic acids as carrier resulted in about a 100 fold higher frequency of transformation with plasmids containing the 2μm origin of replication. Single stranded DNA seems to be responsible for the effect since M13 single stranded DNA, as well as RNA, was effective. Boiled carrier DNA did not yield any increased transformation efficiency using spheroplast formation to induce DNA uptake, indicating a difference in the mechanism of transformation with the two methods.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340712
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  • 8
    Electronic Resource
    Electronic Resource
    The isolation of specific genes from the basidiomycete Schizophyllum commune (1987)
    Springer
    Current genetics 12 (1987), S. 547-554 
    Details
    ISSN: 1432-0983
    Keywords: Schizophyllum commune ; Transformation ; Gene isolation ; Basidiomycetes ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have developed a routine way to isolate genes directly from the basidiomycete fungus, Schizophyllum commune. Plasmid DNA from a genomic gene library was used to isolate five specific genes by complementation of Schizophyllum mutations via transformation. The mutant strains were deficient in the ability to synthesize either adenine (ade2 and ade5), uracil (ural, encoding orotidine-5′-phosphate decarboxylase; OMPdecase), tryptophan (rpl, encoding indole-3-glycerol phosphate synthetase; IGPS) or para aminobenzoic acid (pab1). In each case, Southern analysis revealed that transformation to prototrophy was concomitant with the integration of vector sequence into the genome of the S. commune mutant. Total DNA from transformants was restricted, religated, and used to transform E. coli. Ampicillin resistant plasmids were recovered from E. coli and tested for their ability to transform the corresponding mutant of S. commune. Plasmids complementing the ade2, adeS, pabl, trpl, and ural mutations were recovered.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419565
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  • 9
    Electronic Resource
    Electronic Resource
    Neomycin resistance as a dominantly selectable marker for transformation of the zygomycete Absidia glauca (1987)
    Springer
    Current genetics 12 (1987), S. 625-627 
    Details
    ISSN: 1432-0983
    Keywords: Absidia glauca ; Transformation ; Neomycin resistance ; Actin promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A plasmid (pAmN61) containing the NPT II structural gene (neomycin phosphotransferase) fused to the N-terminal region of a homologous actin gene was used for the transformation of Absidia glauca protoplasts. Neomycin resistant transformants could be selected for on complete medium containing 1.2 mg/ml neomycin sulfate. The physical presence of plasmid DNA in Absidia glauca was demonstrated by retransformation into Escherichia coli and by Southern blot analysis. No integration of plasmid DNA at either one of the two actin loci was observed; Southern blot experiments provide evidence that pAmN61 is autonomously replicated in Absidia glauca.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00368066
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  • 10
    Electronic Resource
    Electronic Resource
    A colony procedure for transformation of Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 21-23 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Transformation ; Plasmid ; Colony ; Polyethylene glycol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid and simple yeast transformation procedure has been developed using colonies on agar plates. Saccharomyces cerevisiae SHY3 cells were picked up from colonies on YPD plates grown freshly or stored at 4 °C and incubated with M13RK9-T DNA at 30 °C for 1–2 h in a solution of Li+, Ca2+, Mg2+, triacetin and polyethylene glycol. About 3,500 transformants were obtained per µg of double stranded M13RK9-T DNA. Unlike the existing spheroplast techniques, single stranded M13RK9-T DNA transformed intact cells below one-hundredth frequency of the duplex form.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365751
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