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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 22 (2000), S. 217-223 
    ISSN: 1573-0603
    Keywords: Flow cytometry ; Muscle cell ; Porcine ; Transfection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A detailed methodology is described for fluorescence-activated cell sorting (FACS) of porcine muscle cells that have been transfected to express green fluorescent protein (GFP). Cells are liberated from porcine skeletal muscle and primary cultures are transfected with DNA encoding GFP. Primary cultures are subjected to immunocytochemistry using a primary muscle-specific monoclonal antibody followed by incubation with a phycoerythrin-conjugated second antibody. Transfected myoblasts aresorted from fibroblasts using forward angle light scatter and ninety degree light scatter, phycoerythrin fluorescence, and GFP fluorescence. These procedures allow for isolation of genetically-engineered porcine muscle cells more rapidly than traditional clonal selection procedures. Consequently, FACS provides porcine myoblast populations that retain the majority of their replicative capacity and are not contaminated with non-myogenic cells.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5079
    Keywords: ascorbic acid ; ascorbate-glutathione cycle ; bean yield ; dehydroascorbate ; ozone ; photosynthesis ; soybean ; vegetative yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We examined the characteristics of ascorbic acid (ASC) level, dehydroascorbate (DHA) level, and the ASC–DHA redox status in the leaflets of two soybean cultivars grown in a field environment and exposed to elevated ozone (O3) levels. These two cultivars, one that preliminary evidence indicated to be O3-tolerant (cv Essex), and one that was indicated to be O3-sensitive (cv Forrest), were grown in open-top chambers during the summer of 1997. The plants were exposed daily to a controlled, moderately high O3 level (≈58 nl l−1 air) in the light, beginning at the seedling stage and continuing to bean maturity. Concurrently, control plants were exposed to carbon-filtered, ambient air containing a relatively low O3 level (≈24 nl l−1 air) during the same period. Elevated O3 did not affect biomass per plant, mature leaf area accretion, or bean yield per plant of cv Essex. In contrast, elevated O3 level decreased the biomass and bean yield per plant of cv Forrest by approximately 20%. Daily leaflet photosynthesis rate and stomatal conductance per unit area did not decrease in either cultivar as a result of prolonged O3 exposure. A 10% lower mature leaflet area in O3-treated cv Forrest plants contributed to an ultimate limitation in long-term photosynthetic productivity (vegetative and bean yield). Possible factors causing cv Essex to be more O3 tolerant than cv Forrest were: 1) mature leaflets of control and O3-treated cv Essex plants consistently maintained a higher daily ASC level than leaflets of cv Forrest plants, and 2) mature leaflets of cv Essex plants maintained a higher daily ASC–DHA redox status than leaflets of cv Forrest plants.
    Type of Medium: Electronic Resource
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