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  • 1
    ISSN: 1617-4623
    Keywords: Bactrocera tryoni ; Excision assay ; Lucilia cuprina ; Transposable element
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plasmid-based excision assays performed in embryos of two non-drosophilid species using the mariner transposable element from Drosophila mauritiana resulted in empty excision sites identical to those observed after the excision of mariner from D. mauritiana chromosomes. In the presence of the autonomous mariner element Mos1, excision products were recovered from D. melanogaster, D. mauritiana and the blowfly Lucilia cuprina. When a hsp82 heat shock promoter-Mos1 construct was used to supply mariner transposase, excision products were also recovered from the Queensland fruitfly Bactrocera tryoni. Analysis of DNA sequences at empty excision sites led us to hypothesise that the mariner excision/repair process involves the formation of a heteroduplex at the excision breakpoint. The success of these assays suggests that they will provide a valuable tool for assessing the ability of mariner and mariner-like elements to function in non-drosophilid insects and for investigating the basic mechanisms of mariner excision and repair.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-6857
    Keywords: Drosophila ; Musca ; recombination ; transgenics ; transposable elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic insect technology will provide opportunities to explore the basic biology of a broad range of insect species in ways that will prove insightful and important. It is also a technology that will provide opportunities to manipulate the genotypes of insects of practical significance to the health and welfare of humans. The Hermes transposable element from the housefly, Musca domestica, is a short inverted repeat-type element related to hobo from Drosophila melanogaster, Ac from Zea mays, and Tam3 from Antirrhinum majus. It has potential to become a versatile and efficient broad host-range insect transformation vector. The ability of Hermes to transpose when introduced into five species of diptera from four divergent families was tested using an in vivo, interplasmid transpositional recombination assay. Hermes was capable of transposing in all species tested, demonstrating that Hermes has a broad host-range. In addition, the rates of transposition were sufficiently high in all species tested to suggest that Hermes will be an efficient gene transfer vector in a wide range of insect species. The Hermes element also revealed a pattern of integration into the target substrate that permitted factors determining integration site selection to be identified. Primary nucleotide sequence of the integration site played a role as did proximity to preferred integration sites and the nucleosomal organization of the target.
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  • 3
    ISSN: 1573-6857
    Keywords: hermit ; hobo ; Lucilia cuprina ; transposable element
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the cloning ofhermit, a member of thehAT family of transposable elements from the genome of the Australian sheep blowfly,Lucilia cuprina. Hermit is 2716 bp long and is 49% homologous to the autonomoushobo element,HFL1, at the nucleic acid level.Hermit has 15 bp terminal inverted repeats that share 10 bp with the terminal inverted repeats ofHFL1. Conceptual translation reveals a 583 residue open reading frame (ORF) that is 64% similar and 42% identical to theHFL1 ORF. However, the sequence of thehermit element contains two frameshifts within the putative ORF, indication thathermit is an inactive element. Analysis ofL. cuprina strains from within and outside Australia suggested thathermit is present as a single copy in all the genomes analysed.
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  • 4
    ISSN: 1573-6857
    Keywords: Drosophila ; Musca ; recombination ; transgenics ; transposable elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic insect technology will provide opportunities to explore the basic biology of a broad range of insect species in ways that will prove insightful and important. It is also a technology that will provide opportunities to manipulate the genotypes of insects of practical significance to the health and welfare of humans. TheHermes transposable element from the housefly,Musca domestica, is a short inverted repeat-type element related tohobo fromDrosophila melanogaster, Ac fromZea mays, andTam3 fromAntirrhinum majus. It has potential to become a versatile and efficient broad host-range insect transformation vector. The ability ofHermes to transpose when introduced into five species of diptera from four divergent families was tested using anin vivo, interplasmid transpositional recombination assay.Hermes was capable of transposing in all species tested, demonstrating thatHermes has a broad host-range. In addition, the rates of transposition were sufficiently high in all species tested to suggest thatHermes will be an efficient gene transfer vector in a wide range of insect species. TheHermes element also revealed a pattern of integration into the target substrate that permitted factors determining integration site selection to be identified. Primary nucleotide sequence of the integration site played a role as did proximity to preferred integration sites and the nucleosomal organization of the target.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 22 (1993), S. 373-384 
    ISSN: 0739-4462
    Keywords: gene transformation ; non-drosophilid ; Drosophila melanogaster ; Anastrepha suspensa ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A P-element mobility excision assay was used to determine if non-drosophilid insects could support P gene vector function. Present studies included the testing of Muscids, Sphaerocerids, and Phorids, none of which were able to support P mobility. A new excision indicator plasmid was developed allowing the detection and recovery of virtually all P-element excision products. The frequency and sequence analysis of excision products from Drosophila melanogaster and another drosophilid, Chymomyza procnemis, indicated both quantitative and qualitative differences in the activity of transposase. The quantitative relationships observed in the original assay were maintained, and qualitative differences in transposase activity were reflected in the sequence of the empty donor sites. The results suggest that host factors are involved in cutting and ligating P-element DNA during excision, with transposase facilitating these processes. Possible limitations on P mobility by abnormal transpoase transcript processing were tested in Anastrepha suspensa using transposase-encoding plasmids having deleted intron sequences. A transposase cDNA supported normal P excision in D. Melanogaster, and a low level of mobility in A. suspensa. Possible applications of gene transfer in insects are presented, in particular methods to genetically sterilize and sex insects for the sterile-insect technique. © 1993 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
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