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  • 1
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The plasma membrane surface proteins of intact somatic (leaf) and reproductive (pollen, generative cell or sperm cell) protoplasts of lily (Lilium longiflorum) and rapeseed (Brassica napus cv. Midas) were compared after probing with N-hydroxysuccinimido- (NHS) or sulfo-NHS-biotin. The plasma membranes of intact protoplasts are impermeable to these biotin probes, which bind covalently to the free amino groups of surface proteins. Enzyme-labelled streptavidin was used to detect membrane proteins after separation by SDS-PAGE and western blotting. In lily, six proteins specific to the surface membrane of leaf protoplasts were identified varying from 25–64 kDa, three proteins to pollen protoplasts in the range 35–64 kDa and two proteins to generative cell protoplasts, 63 and 67 kDa. In rapeseed leaf protoplasts, seven proteins in the range 22–69 kDa were detected, while in the sperm enriched fraction five proteins were present in the same kDa range. The proteins identified as membrane specific for generative cell protoplasts of lily have been isolated and were used as antigens for monoclonal antibody production. Preliminary results indicate the successful production of antibodies to surface antigens. These antibodies will be used to localise surface specific epitopes which are likely to be involved in cell-cell recognition at fertilization.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 12 (1999), S. 125-126 
    ISSN: 1432-2145
    Keywords: Key words Gene regulation ; Ory s 1 ; Oryza sativa ; Pollen allergen ; Promoter ; Rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5028
    Keywords: Anther ; Brassica napus ; Brassica rapa ; Ca2+-binding protein ; cDNA cloning ; pollen allergen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Thirteen cDNA clones encoding IgE-binding proteins were isolated from expression libraries of anthers of Brassica rapa L. and B. napus L. using serum IgE from a patient who was specifically allergic to Brassica pollen. These clones were divided into two groups, I and II, based on the sequence similarity. All the group I cDNAs predicted the same protein of 79 amino acids, while the group II predicted a protein of 83 amino acids with microheterogeneity. Both of the deduced amino acid sequences contained two regions with sequence similarity to Ca2+-binding sites of Ca2+-binding proteins such as calmodulin. However flanking sequences were distinct from that of calmodulin or other Ca2+-binding proteins. RNA-gel blot analysis showed the genes of group I and II were preferentially expressed in anthers at the later developmental stage and in mature pollen. The recombinant proteins produced in Escherichia coli was recognized in immunoblot analysis by the IgE of a Brassica pollen allergic patient, but not by the IgE of a non-allergic patient. The cDNA clones reported here, therefore, represent pollen allergens of Brassica species.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-5028
    Keywords: Generative cells ; gene expression ; Lilium longiflorum ; pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Metabolic labelling with [35S]-methionine demonstrated that generative cells ofLilium longiflorum possess their own set of mRNA and are capable of synthesising proteins independently from the vegetative cell. The isolated generative cells synthesised ten proteins, of which six were unique to these specialised cells. Isolation of generative cells from pollen grains after [35S]-methionine labelling resulted in an identical protein profile, therefore the synthesis of these proteins was not due to isolation shock. Addition of cycloheximide, abolished TCA-precipitable counts, whilst actinomycin D had no qualitative effect on the observed protein profile, indicating active translation of pre-existing mRNAs by the generative cells.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 39 (1999), S. 607-614 
    ISSN: 1573-5028
    Keywords: generative cell ; histone ; Lilium longiflorum ; male gamete ; pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Formation of the generative cell in flowering plants initiates the male spermatogenesis pathway which eventually culminates in the process of double fertilization. Little is known about the molecular mechanisms operative in the generative cell. Here, we report the isolation and characterization of cDNA clones encoding generative cell-specific histones, gcH2A and gcH3. In situ hybridization analysis revealed that both genes are activated after formation of the generative cell and accumulation of transcripts increases progressively during generative cell maturation. These results suggest that gcH2A and gcH3 mRNAs are products of transcriptional activation of the generative cell nucleus and thus provide the first evidence for male germ line cell-specific gene expression in flowering plants.
    Type of Medium: Electronic Resource
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