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  • 1
    Electronic Resource
    Electronic Resource
    Crystallographic studies of the mechanism of xylose isomerase (1989)
    s.l. : American Chemical Society
    Biochemistry 28 (1989), S. 7289-7297 
    Details
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/bi00444a022
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  • 2
    Electronic Resource
    Electronic Resource
    Concomitant expression of E. coli cytosine deaminase and uracil phosphoribosyltransferase improves the cytotoxicity of 5-fluorocytosine (1998)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 167 (1998), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The prodrug activation system formed by the E. coli codA gene encoding cytosine deaminase (CD) and 5-fluorocytosine (5-FC) developed for selective cancer chemotherapy suffers from a sensitivity limitation in many tumour cells. In an attempt to improve the CD/5-FC suicide association, we combined the E. coli upp gene encoding uracil phosphoribosyltransferase (UPRT) with codA gene to create the situation prevailing in E. coli, a bacterium very efficient in metabolising 5-FC. The constitutive expression of the two genes cloned on an E. coli-animal cell shuttle plasmid either in a linked or in a fused configuration was evaluated in E. coli strains selected and engineered to mimic the 5-FC metabolism encountered in mammalian cells. The simultaneous expression of codA and upp genes generated a cooperative effect resulting in a dramatic increase in 5-FC sensitivity of cells compared to the expression of codA alone. Furthermore, it was shown that the association of UPRT with CD facilitated the uptake of 5-FC, in the situation where the drug penetrates cells by passive diffusion as in mammalian cells, by directly channeling 5-fluorouracil, the product of CD, to 5-fluoroUMP, the product of UPRT.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1998.tb13205.x
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  • 3
    Electronic Resource
    Electronic Resource
    Characterization and Cloning of Glucose Isomerase from Streptomyces violaceoniger (1987)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 501 (1987), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1987.tb45691.x
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  • 4
    Electronic Resource
    Electronic Resource
    Transformation of Aspergillus flavus: construction of urate oxidase-deficient mutants by gene disruption (1992)
    Springer
    Current genetics 21 (1992), S. 447-453 
    Details
    ISSN: 1432-0983
    Keywords: Aspergillus flavus ; Urate oxidase ; Transformation ; Gene disruption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A transformation procedure based on the complementation of a genetic defect was developed using a nitrate reductase-deficient mutant of Aspergillus flavus. The initial transformation efficiency was improved 40-fold by combining factors in a planned experimental program. Although low, this transformation rate was sufficient to obtain transformants in which the urate oxidase-encoding gene (uaZ) was disrupted in a gene replacement experiment. These new uaZ- strains were unable to utilize uric acid as the unique nitrogen source and could be reversed directly to the wild-type phenotype in second order transformation experiments using a urate oxidase-expressing vector.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00351654
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  • 5
    Electronic Resource
    Electronic Resource
    High efficiency transformation of Tolypocladium geodes conidiospores to phleomycin resistance (1991)
    Springer
    Current genetics 20 (1991), S. 309-314 
    Details
    ISSN: 1432-0983
    Keywords: Phleomycin resistance ; Transformation ; Fungi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A convenient and efficient transformation system has been developed for the filamentous fungus Tolypocladium geodes. In contrast to most of the commonly described techniques requiring prior preparation of protoplasts or spheroplasts, this method leads to high efficiency transformation of T. geodes conidiospores following moderate lytic enzyme treatment. Competent cells so obtained are still resistant to osmotic pressure and can be stored frozen without loss of viability. The highest transformation frequency (3-5x103 transformants per μg of DNA) was obtained with plasmid pUT737 containing the Sh ble gene conferring phleomycin resistance under the control of a strong promoter isolated from Trichoderma reesei. Southern hybridization revealed multiple integration sites of plasmid DNA into the T. geodes nuclear DNA despite the absence of homology between the transforming DNA and the recipient genome. Instability could not be detected for the phleomycin® phenotype during more than five generations of mitotic growth under non-selective conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318520
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  • 6
    Electronic Resource
    Electronic Resource
    Production of extracellular pectinase enzymes by a mutant (Pol6) of Penicillium occitanis (1990)
    Springer
    Applied microbiology and biotechnology 34 (1990), S. 308-312 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Penicillium occitanis strain Pol6, a mutant developed for hyperproduction of cellulase and pectinase enzymes was used for the study of extracellular pectinase production when pectins from different sources (apple and citrus) and with varying degree of esterification (DE) were used as inducers. Highly esterified citrus pectins were found to be suitable substrates for polygalacturonase, pectinase and pectin methyl esterase production, while low esterified citrus pectin favoured pectin lyase (PL) production. Apple pectins induced other hydrolytic enzymes (e.g., β-1,3-glucanase, β-glucosidase, β-galactosidase), in addition to pectolytic enzymes. Moreover, the combination of high and low esterified citrus pectins induced the production of a complete pectinase complex. The extent of degradation of the substrate and the affinity for PL decreased with decreasing DE irrespective of the source. There was no evidence of PL activity in this strain. No significant effect of cations (Ca++, Mn++, Na+) on PL activity was observed. However, EDTA (100 mm) inhibited 50% of the activity, when tested on highly esterified (rapid set citrus) pectin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00170048
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  • 7
    Electronic Resource
    Electronic Resource
    Phleomycin resistance as a dominant selectable marker in CHO cells (1988)
    Springer
    Somatic cell and molecular genetics 14 (1988), S. 243-252 
    Details
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The Tn5 and the Streptoalloteichus hindustanus (Sh) ble genes conferring resistance to bleomycin-phleomycin antibiotics have been cloned into a mammalian vector under the RSV-LTR promoter. The resulting plasmids, pUT506 and pUT507 respectively, were used to transfect CHO cells by either the calcium phosphate or the recently described polybrene-DMSO method. Phleomycin- or bleomycin-resistant clones arose with a higher frequency after transfection with pUT507, and pUT507 transfectants were more resistant to both antibiotics than pUT506 transfectants. Phleomycin resistance in pUT507 transfectants was stable and associated with integration of plasmid sequences in genomic DNA. The Sh ble gene, which confers a dominant phleomycin-resistance phenotype, should provide a useful transferable selectable marker in CHO cells as well as in other animal cell lines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01534585
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  • 8
    Electronic Resource
    Electronic Resource
    Phleomycin resistance as a dominant selectable marker for plant cell transformation (1989)
    Springer
    Plant molecular biology 13 (1989), S. 365-373 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium ; binary vectors ; bleomycin ; phleomycin ; selective markers ; tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tobacco cells are sensitive to bleomycin and phleomycin. The Tn5 and the Streptoalloteichus hindustanus (Sh) bleomycin resistance (‘Ble’) genes conferring resistance to these antibiotics have each been inserted into two plant expression vectors. They are flanked by the nopaline synthase (nos) or the cauliflower mosaic virus (CaMV) 35S promoters on one side, and by the nos polyadenylation signal on the other. These four chimaeric genes were introduced into the binary transformation vector pGA 492, which were thereafter mobilized into Agrobacterium tumefaciens strain LBA 4404. The resulting strains were used to transform Nicotiana tabacum cv. Xanthi nc using the leaf disc transformation procedure. In all cases, phleomycin- and bleomycin-resistant tobacco plants were regenerated from transformed cells under selective conditions; however the highest frequency of rooted plants was obtained when transformation was carried out with the ‘Sh Ble’ gene under the control of the 35S promoter. Phleomycin resistance was stably transmitted to sexual offspring as a dominant nuclear trait as confirmed by Southern blotting.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00015548
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  • 9
    Electronic Resource
    Electronic Resource
    Phleomycin resistance encoded by the ble gene from transposon Tn 5 as a dominant selectable marker in Saccharomyces cerevisiae (1987)
    Springer
    Molecular genetics and genomics 207 (1987), S. 342-348 
    Details
    ISSN: 1617-4623
    Keywords: Phleomycin ; Tn5 ; Yeast transformation ; Iso-1-cytochrome C promoter ; Dominant markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Phleomycin, a water-soluble antibiotic of the bleomycin family is as effective against Saccharomyces cerevisiae cells as against Escherichia coli cells. The ble gene of transposon Tn5, which confers resistance to phleomycin, was inserted in place of the iso-1-cytochrome C (CYC1) gene on an autonomously replicative multicopy E. coli-yeast shuttle plasmid. Higher resistance levels are obtained in S. cerevisiae when the region immediately upstream from the initiation codon conforms to the nucleotide sequence stringencies observed in almost every yeast gene. The expected regulation pattern of the whole CYC1 promoter confers different phleomycin resistance levels to the cell under varying physiological conditions. Partial deletions in the CYC1 promoter lead to changes in the resistance level of cells which are mostly accounted for by the removal of known positive and negative regulatory elements. Some of the vector constructions allow direct selection of phleomycin-resistant transformants on rich media.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331599
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  • 10
    Electronic Resource
    Electronic Resource
    Development of a transformation system for the thermophilic fungus Talaromyces sp. CL240 based on the use of phleomycin resistance as a dominant selectable marker (1992)
    Springer
    Molecular genetics and genomics 234 (1992), S. 489-493 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Shotgun cloning ; Filamentous fungi ; Promoter sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A transformation system for the thermophilic cellulolytic fungus Talaromyces sp. CL240 has been developed, using the phleomycin resistance gene from Streptoalloteichus hindustanus (Sh ble) as a dominant selectable marker. The plasmids (pAN8-1 and pUT720) carrying the Sh ble gene under the control of the Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter, allowed selection of phleomycin-resistant transformants. A new promoter sequence cloned from chromosomal DNA of Trichoderma reesei (pUT737) was also able to drive efficient expression of the Sh ble gene in Talaromyces sp. CL240, resulting in the selection of transformants that were highly resistant to phleomycin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00538710
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