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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Development genes and evolution 210 (2000), S. 623-629 
    ISSN: 1432-041X
    Keywords: Keywords Germline transformation ; Green fluorescent protein ; Pax-6 ; Transposable elements ; Transposon mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The efficiency of transposon-mediated germline transformation is dependent on the transposon mobility in the host embryo, and on the detectability of the used transformation marker. Therefore, high susceptibility of the transformation marker to position effect suppression is a disadvantage. Here we present data that the eye-specific expression of green fluorescent protein, driven by the 3xP3-EGFP marker, outperforms the commonly used ”mini”-white transformation marker in Drosophila germline transformation experiments: 3xP3-EGFP is more sensitive than ”mini”-white in identifying transgenic individuals and reacts differently to position effect suppression. Therefore, 3xP3-EGFP offers an ideal marker for applications in functional genomics where as many gene loci as possible should be targeted in the genome of a specific organism, for example, as intended in the Drosophila gene disruption project. Furthermore, we give a detailed description of the embryonic and larval expression mediated by the 3xP3-EGFP marker. These pre-adult expression patterns, and the potentially universal applicability of the transformation marker also offer additional advantages for selecting transgenic individuals in organisms other than Drosophila. This will be of great interest to the field of evolutionary developmental biology and to modern pest management programs.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Development genes and evolution 210 (2000), S. 630-637 
    ISSN: 1432-041X
    Keywords: Keywords Green fluorescent protein ; Pax-6 ; Transformation marker ; Transposable elements ; Transposon mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Genetic manipulation of a series of diverged arthropods is a highly desirable goal for a better understanding of developmental and evolutionary processes. A major obstacle so far has been the difficulty in obtaining marker genes that allow easy and reliable identification of transgenic animals. Here, we present a versatile vector set for germline transformation based on the promiscuous transposons mariner, Hermes and piggyBac. Into these vectors, we introduced a potentially universal marker system that is comprised of an artificial promoter containing three Pax-6 homodimer binding sites. This promoter drives strong expression of spectral variants of the enhanced green fluorescent protein (EGFP) in larval, pupal, and adult photoreceptors. Using special filter sets, the yellow (EYFP) and cyan (ECFP) variant are fully distinguishable and therefore represent a separable pair of markers. Furthermore, we adapted a simple plasmid-based transposition assay system to enable quick functional tests of our vectors in different arthropod species before employing them in more laborious germline transformation experiments. Using this system we demonstrate that our vectors transpose in both Drosophila melanogaster and Drosophila virilis.
    Type of Medium: Electronic Resource
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  • 3
    Publication Date: 2000-11-21
    Print ISSN: 0949-944X
    Electronic ISSN: 1432-041X
    Topics: Biology
    Published by Springer
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  • 4
    Publication Date: 2000-11-21
    Print ISSN: 0949-944X
    Electronic ISSN: 1432-041X
    Topics: Biology
    Published by Springer
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