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  • transformation
  • Springer  (12)
  • Cambridge University Press (CUP)
  • PANGAEA
  • 1985-1989  (10)
  • 1955-1959  (2)
  • 1935-1939
Collection
Publisher
  • Springer  (12)
  • Cambridge University Press (CUP)
  • PANGAEA
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Year
  • 1
    ISSN: 1573-5028
    Keywords: Agrobacterium ; gene expression ; legumin (Pisum) ; Nicotiana ; seed storage protein ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 3.4-kilobase genomic DNA fragment from Pisum sativum L. containing the LegA gene, which encodes a major legumin storage protein, was transferred to Nicotiana plumbaginifolia using an Agrobacterium tumefaciens strain containing the Bin 19 binary vector system. Northern hybridisation analysis of legA-transformed plants demonstrated that legumin-specific RNA was present in developing seeds but not in developing leaves. Legumin protein was immunologically detected in the mature seeds of legA-transformed plants, and was present as the correct-size protein composed of disulphide-bonded polypeptides. It is concluded that the transferred pea genomic fragment contains all the information necessary for seed-specific expression of the legA gene, and for correct processing of the primary transcript and the precursor legumin protein.
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  • 2
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; Beta vulgaris ; sugar beet ; regeneration ; shooter mutants ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Beta vulgaris plants were found to be susceptible to Agrobacterium tumefaciens strains carrying octopine Ti-plasmids after wounding of GA3 elongated stems or of hypocotyls. Tumors could be isolated and cultured aseptically. The tumor marker, octopine synthase (Ocs) activity, was present demonstrating the applicability of the Agrobacterium system for transfer of genetic information. For the production of transgenic plants two procedures were tested: inoculation of explants derived from cotyledons and hypocotyls of two weeks old seedlings and a leaf-disc procedure. The first method yielded both octopine positive calli as well as shoot regeneration on the six genotypes tested. In most cases, regeneration occurred from pre-existing, non-transformed meristems. The presence of Ocs activity could not be demonstrated in these shoots, although in one case octopine positive callus was formed at the base of the shoot, suggesting a chimeric structure of the plantlet or T-DNA genes, which were silent within the shoot and became active again in proliferating callus. The leaf-disc method did not give rise to direct or indirect regeneration, but transformed callus proliferated on the leaf edges. Optimal transformation frequencies were dependent on B. vulgaris genotype and Agrobacterium strain. The use of Agrobacterium shooter mutants or strains carrying an isolated cytokinin gene in order to influence endogenous phytohormone ratios did not result in the formation of shoots nor did it increase levels of regeneration in the first method. Further optimization is in order and in progress.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Bulletin of experimental biology and medicine 105 (1988), S. 389-392 
    ISSN: 1573-8221
    Keywords: immortalization ; oncogenes ; transfection ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 12 (1989), S. 127-133 
    ISSN: 1573-0603
    Keywords: polyethylene glycol ; transformation ; protoplasts ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Polyethylene glycol can be used to induce DNA uptake into plant protoplasts. Procedures for isolation, culture and transformation ofN. tabacum protoplasts are described and can be adapted for other dicot and monocot species. Criteria for proof of transformation are discussed.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 13 (1989), S. 151-161 
    ISSN: 1573-5028
    Keywords: β-glucuronidase (gusA) gene ; maize ; protoplasts ; stable co-transformation ; transformation ; Zea mays L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient co-transformation protocol using polyethylene glycol was developed for Zea mays L. (cv. A188 × BMS) protoplasts isolated from suspension culture cells. Co-transformation was accomplished by using plasmid constructions containing β-glucuronidase (gusA) or neomycin phosphotransferase (neo) gene coding sequences; both were under control of the CaMV 35S promoter. Protoplast culture and transformation conditions were optimized to assure efficient recovery of transformed cells. The overall efficiency of transformation was 1 × 10−4 (calculated per viable protoplast plated). Among kanamycin-resistant lines, 50% showed a high level of GUS activity (above one unit). Southern blot hybridization confirmed the presence of numerous gusA and neo coding sequences in the maize genome. In two analyzed lines, integrated sequences appeared to be organized in tandem head-to-tail repeats. Results also indicated that the integrated sequences were partially methylated.
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  • 6
    ISSN: 1573-5028
    Keywords: herbicide ; mutant ; photosystem II ; psbA gene ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutations conferring herbicide resistance in 3 mutant strains of the cyanobacterium Synechocystis 6714 have been characterized by gene cloning and sequencing. The mutants display very different phenotypes: DCMU-IIA is DCMU-resistant and atrazine-resistant, DCMU-IIB is DCMU-resistant and atrazine-sensitive, and Az-V is DCMU-sensitive, atrazine-resistant and presents particular photoinhibition properties. These mutants were originally obtained either by one-step selection (DCMU-IIA) or by two-step selection (DCMU-IIB and Az-V). psbA copies carrying herbicide resistance have been identified by transformation experiments as psbAI in all cases. Sequences of the psbAI copy of each mutant have been compared to the wild-type sequence. In the single mutant DCMU-IIA, a point mutation at codon 264 (Ser→Ala) results in resistance to both DCMU and atrazine. In the double mutants DCMU-IIB and Az-V, two point mutations were found. DCMU-IIB was derived from DCMU-IIA and had acquired a second mutation at codon 255 (Phe→Leu) resulting in a slight increase in DCMU resistance and complete abolition of atrazine resistance. Az-V contains two changes at codons 211 (Phe→Ser) and 251 (Ala→Val) resulting in high atrazine resistance but only slight DCMU resistance.
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  • 7
    ISSN: 1573-5060
    Keywords: Hordeum vulgare ; isolated microspores ; particle bombardment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A highly regenerable, isolated microspore system for barley, Hordeum vulgare L. cv. Igri, has been developed which is amenable to transformation studies using particle bombardment. The system allows DNA to be delivered to microspores at the single cell stage and both transient and stable transformation events have been demonstrated. The potential advantages of using isolated microspores as the target tissue in routine transformation systems are discussed.
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  • 8
    ISSN: 1573-5060
    Keywords: alfalfa ; alpha-amylase ; field performance ; manganese-dependent lignin peroxidase ; Medicago sativa ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Transgenic alfalfa plants expressinBacillus licheniformis alpha-amylase and mangaese-dependent lignin peroxidase (Mn-P) from Phanerochaete chrysosporium were produced using the Agrobacterium tumefaciens transformation system. In each case, there was a range of expression of the introduced gene among independent transgenic plants. Plants producing alpha-amylase showed no alteration of phenotype. Production of Mn-P in alfalfa, howeven, in most cases adversely affected plant growth and development. Affected plants were stunted with yellowing foliage, but survived and produced seed. Results from field trials showed that Mn-P production in transgenic alfalfa reduced dry matter yield and plant height. The extent of these symptoms and yield reduction was, for the most part, related to the level of foreign protein production as estimated by Western analysis. Field data from transgenic plants expressing alpha-amylase showed that there was no effect of foreign protein production on plant performance. Expression of Mn-P was shown to segregate in sexual progeny derived from transgenic plants.
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  • 9
    ISSN: 1573-5028
    Keywords: Agrobacterium rhizogenes ; binary vector ; kanamycin resistance ; transformation ; secondary metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Co-transfer of Agrobacterium rhizogenes T-DNA and T-DNA from the A. tumefaciens binary vector pBin19 (Bevan, 1984) was studied in detail using Nicotiana rustica. High frequencies of co-transfer of T-DNA's were observed, even when no selection pressure was exerted. Increased levels of pBin19 T-DNA were found in hairy root cultures with selection at higher levels of kanamycin sulphate (50–200 μg ml−1). Several other species were also transformed by A. rhizogenes carrying pBin19 and A. rhizogenes harbouring a different binary factor, pAGS125 (Van den Elzen et al., 1985), was used to transform N. rustica hairy roots to confer hygromycin B resistance.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 10 (1986), S. 117-123 
    ISSN: 1573-0603
    Keywords: growth factors ; DNA synthesis ; cancer ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid mitogenic assay suitable for the detection of transforming growth factors in the extracts of tissues or cells or in the medium conditioned by tumor cells in vitro is described. The method utilizes a nontumorigenic mouse embryo cell line (AKR-2B cells) maintained in serum-free conditions. Three classes of growth factors can be distinguished using this assay.
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