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  • 1
    Electronic Resource
    Electronic Resource
    Production of fertile transgenic maize by electroporation of suspension culture cells (1994)
    Springer
    Plant molecular biology 24 (1994), S. 51-61 
    Details
    ISSN: 1573-5028
    Keywords: Zea mays L. ; transformation ; electroporation ; bar ; phosphinothricin acetyltransferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fertile, transgenic maize plants were generated by electroporation of suspension culture cells that were treated with a pectin-degrading enzyme. Electroporation of cells from two different suspension cultures, one derived from A188 X B73 and one derived from a B73-related inbred, with a plasmid containing the bar gene, resulted in high-frequency recovery of stably transformed callus lines. Plants were regenerated from thirteen transformed callus lines and transmission of bar to progeny was demonstrated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040573
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  • 2
    Electronic Resource
    Electronic Resource
    Stress responses in alfalfa (Medicago sativa L.). 8. Cis-elements and trans-acting factors for the quantitative expression of a bean chalcone synthase gene promoter in electroporated alfalfa protoplasts (1991)
    Springer
    Plant molecular biology 16 (1991), S. 877-890 
    Details
    ISSN: 1573-5028
    Keywords: chalcone synthase promoter ; electroporation ; elicitor ; transient gene expression ; alfalfa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A chimeric gene consisting of a bean (Phaseolus vulgaris L.) chalcone synthase (CHS) promoter fused to a bacterial chloramphenicol acetyltransferase (CAT) reporter gene was strongly expressed, and further induced by fungal elicitor, when electroporated into alfalfa (Medicago sativa L.) suspension cell protoplasts. Functional analysis of 5′ deletions of the CHS promoter-CAT construct in these protoplasts indicated that the region between −326 and −130 contained both activator and silencer elements. Co-electroporation experiments confirmed that these cis-acting elements were binding sites for functionally active trans factors. In vitro DNase I footprinting revealed four potential binding sites for alfalfa suspension cell nuclear proteins between positions −326 and −130 of the CHS promoter. These sites mapped to regions shown to contain functional cis-acting elements on the basis of the deletion analysis. Three of these sites mapped to previously identified binding sites for bean nuclear proteins. Competition gel retardation analysis using oligonucleotide probes containing binding site sequences revealed sequence-specific binding of alfalfa nuclear proteins to an AT-rich element and a putative GT-1 factor consensus binding sequence. Our results define cis elements and their cognate trans factors functionally active in determining the quantitative expression of a defense response gene in a heterologous transient expression system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00015079
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  • 3
    Electronic Resource
    Electronic Resource
    Rapid identification and characterization of peroxisomal assembly mutants in Yarrowia lipolytica (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 9 (1993), S. 507-517 
    Details
    ISSN: 0749-503X
    Keywords: Peroxisome ; immunofluorescence ; PTS-1 ; electroporation ; yeast ; targeting ; biogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe the isolation and characterization of peroxisomal assembly mutants in the genetically manipulable yeast Yarrowia lipolytica (pay mutants). These mutants were initially identified as oleic acid-non-utilizers by their inability to grow on oleic acid, the utilization of which requires peroxisomal β-oxidation enzymes. Identification of a subset of oleic acid-non-utilizers as pay mutants was obtained by a rapid immunofluorescence procedure using antibodies to the peroxisomal targeting signal Ser-Lys-Leu-CO2H. Punctate structures characteristic of peroxisomes were not detected in pay mutants using this technique. This rapid identification by immunofluorescence should be generally applicable to the selection of peroxisomal assembly mutants in other yeasts. To take advantage of the pay mutant system, we constructed a genomic library in the autonomously replicating vector pINA445 and developed an efficient and rapid electroporation procedure for the functional complementation of these mutants. We have been successful in functionally complementing two independent pay mutants. Molecular analysis of these and other complementing genes will allow for characterization of some of the cellular elements involved in peroxisomal assembly.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320090506
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    Paper (German National Licenses)
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