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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 51 (1981), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Rosa Paul's Scarlet’cell suspension cultures were used as a test system for working out a method of viability and drug-sensitivity determination based on plating efficiency. High plating efficiencies (80–95%) were obtained on a simple synthetic medium when aggregates of a mean size of c. 100 cells/unit from exponential phase cultures were plated at a density of 1500 units/plate in the middle layer (5 ml) of three layers of the agar-solidified medium (total = 30 ml). This 3-layer plating technique produces homogeneous colony growth and simplifies the microscopical evaluation of plating efficiencies. The reduction of plating efficiencies seen when the smaller aggregates of stationary phase cultures were plated was mainly due to low cell density and could be overcome by enriching the medium with various supplements. Reconstitution experiments using mixtures of inactivated and non-inactivated aggregates demonstrated that plating efficiency can be taken as a goodmeasure of viability. The described plating technique was found to be more sensitive and reliable compared to two other methods for determining p-fluorophenylalanine-sensitivity of Rosa cells.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 48 (1980), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Sustained divisions of protoplast-derived cells obtained from cell suspension cultures of Rosa‘Paul's Scarlet’ leading to colony and callus formation was achieved. The rather low plating efficiencies observed for agar-plated protoplasts were partly due to early arrests in further development of dividing protoplast-derived cells and cell clusters. Successful cultivation of dividing protoplast-derived cells was particularly dependent upon frequent subculturing of the precultures and on an efficient procedure for viable protoplast isolation. Colony formation was largely independent of medium composition.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1072-8368
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Sir—Grb2 is a 25,000 Mr ‘adaptor’ protein comprising one SH2 and two SH3 domains1,2. It is found in the cytoplasm of cells in complex with the Son of sevenless (Sos) protein. The complex is formed through the interaction of the Grb2 SH3 domains with the proline-rich region of ...
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: Cryopreservation ; Drug resistance (variant strains) ; Feedback sensitivity ; Isoleucine ; l-Threonine deaminase ; O-Methylthreonine ; Rosa (cell culture)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract O-Methylthreonine (OMT) inhibits the growth of plated Rosa cells (ID50≃6·10-6M). Isoleucine is able to reverse efficiently and specifically this OMT toxicity. From OMT-resistant colonies occurring at a frequency of 1.58·10-7 variants per cell plated at 10-4M OMT, the variant strains OMTR-1 and OMTR-2 were isolated, cloned via protoplasts and characterized. Both variants were ten times more resistant to OMT than the wildtype and were cross-resistant to another isoleucine analog, dl-4-thiaisoleucine. The resistant variants retained their resistance after storage for three years in liquid nitrogen. Both resistant strains were stable for several months when subcultured in the absence of OMT although it was shown in a reconstitution experiment that wildtype cells overgrow OMTR-2 variant cells if co-cultivated for many passages in drug-free medium. One case of instability was observed upon long-term subculturing in drug-free medium: the strain OMTR-1D* partially lost phenotypic properties. Resistance to OMT was followed qualitatively by a new method based on inhibition-zone formation in cell suspensions plated in agar medium. The OMT-resistant variants showed a reduction in sensitivity of the enzyme l-threonine deaminase to feedback inhibition by isoleucine, a decreased stability of l-threonine deaminase when stored at-18°C or incubated at +55°C and a two- to threefold increase of the free isoleucine pool within the cells. The genetical events and the biochemical mechanisms which might lead to the observed stable and biochemically defined character are discussed with particular reference to the high ploidy level of the Rosa cell line.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using Agrobacterium rhizogenes, roots were induced on explants of 24 different Solanaceae species and established as in vitro cultures. Some of the root clones produced tropane alkaloids at levels similar to roots of the corresponding intact plants and maintained these levels over several passages.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0983
    Keywords: Gene cluster ; Tryptophan ; Schizosaccharomyces pombe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 65 trp1− alleles of Schizosaccharomyces pombe have been analysed for their interallelic complementation pattern, suppressibility by nonsense suppressors and position of the corresponding mutation site on an intragenic map of the trp1 locus. In addition to the three complementation classes previously described (Schweingruher and Dietrich 1973) as defective in phosphoribosylanthranilate isomerase (trp1A), indole glycerolphosphate synthetase (trp1B) and anthranilate synthetase (trp1C), two new complementation classes, trp1BC and trp1ABC, were found. The former is represented by a single allele which can only complement trp1A mutants. The latter is represented by six alleles which fail to complement tester mutants of the trp1A, trp1B or trp1C class. Classes trp1A, trp1B and trp1C correspond to mutations in three nonoverlapping regions mapping in the order trp1A, trp1B and trp1C All the alleles of the trp1BC and trp1ABC classes correspond to mutations in the trp1C region. Nonsense alleles of the opal (UGA) or ochre (UAA) type were found in the trp1A (2 alleles out of 17) and trp1ABC (5 alleles out of 6) classes only. These data indicate that the trp1 locus is transcribed as a single messenger RNA with transcription starting from the trp1C region. This messenger is probably translated in a single, multifunctional polypeptide, or at most in two polypeptides coded for by the trp1B-trp1C and by the trp1A regions. In addition, the polar effect of nonsense mutations in the trp1C region is cancelled by rare spontaneous mutations occuring at or very near the trp1 locus, which may act by creating an internal single for the initiation of transcription and/or translation.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2048
    Keywords: Auxotroph ; Haploid ; Hyoscyamus ; Nitrate reductase ; Protoplasts ; Variants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A population of 3070 clones derived from N-methyl-N′-nitro-N-nitrosoguanidine (MNNG)-treated mesophyll protoplasts of haploid Hyoscyamus muticus was tested for amino-acid auxotrophy without enrichment. One clone (MA-2) was stably and specifically dependent on casein hydrolysate and could be fed also by a number of single amino acids or by other reduced nitrogen sources. MA-2 was found to be chlorate resistant and devoid of in vivo nitrate reductase activity under inductive conditions. Permissive and restrictive growth conditions for MA-2 were investigated more closely and media were found promoting morphogenesis. Selection and testing of clones were complicated by an unspecific growth stimulation of some wild type cultures by amino acids, thiamine and m-inositol.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 172 (1979), S. 233-241 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The complex locus aro3 of Schizosaccharomyces pombe was subjected to genetical fine structure analysis. By comparing the complementation map and the meiotic recombination map, the aro3 locus could be subdivided into the five adjacent subregions A, B, C, D and E. Out of 115 aro3 alleles, 26 nonsense alleles and 30 missense alleles could be identified by the criteria of nonsense suppressor sensitivity and leakiness, respectively. Most alleles with a pleiotropic complementation pattern are of the nonsense type. We conclude from the polarity of the complementation patterns characterising the nonsense alleles that the translation direction proceeds from subregion A to subregion E. Antipolar effects in complementation are more frequent than in the analogous system of the arom gene cluster of Neurospora crassa.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 3 (1984), S. 111-122 
    ISSN: 1573-5044
    Keywords: Rosa cell suspension culture ; disk method ; inhibition zone ; drug sensitivity ; L-ethionine ; mercuric chloride ; diffusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When paper disks carrying small volumes of highly concentrated drugs were placed on the suface of the medium in plant cell culture plates, diffusion of the drugs led to a circular area of non-dividing cells (an inhibition zone) around the disks. Out of 63 drugs tested 37 were inhibitory and 15 of these produced a clear inhibition zone. Drug concentrations could be estimated by measuring the inhibition zone diameter. Cell growth and drug diffusion were analysed and the influence of several variables on inhibition zone formation studied. Inhibition zones occured with cultures of Zea mays, Acer pseudoplatanus, Daucus carota and Hyoscyamus muticus and protoplast-derived cells of Rosa. Possible applications of the method in plant cell genetics and physiology are discussed.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 3 (1984), S. 123-130 
    ISSN: 1573-5044
    Keywords: Rosa cell suspension culture ; disk method ; exhibition zone ; amino acid analog ; reversion of drug toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Growth inhibition of plated cells by antimetabolites present in the culture medium was alleviated by corresponding metabolites supplied on filter paper disks placed on the surface of the medium. The metabolites diffused into the medium producing a halo of growing cells (an exhibition zone). Exhibition zones were recorded for 15 pairs of compounds and with cell cultures of Rosa, Acer pseudoplatanus, Daucus carota, Hyoscyamus muticus and Zea mays. The interaction between the compounds could be quantified by measuring exhibition zone diameters and the method was partially optimised for L-ethionine/L-methionine and p-fluorophenylalanine/L-phenylalanine. The exhibition zone method was compared to two other methods, measurement of growth yield and plating efficiency, for demonstrating reversion of growth inhibition.
    Type of Medium: Electronic Resource
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