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  • 1
    ISSN: 1617-4623
    Keywords: Homologous recombination ; Protoplast transformation ; β-Glucuronidase ; Maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The requirements for homologous recombination between plasmid DNA molecules have been studied using the PEG (polyethylene glycol)-mediated transformation system of maize (Zea mays L.) protoplasts coupled with the transient expression assay for β-glucuronidase (GUS). Two plasmids were introduced into maize protoplasts; one plasmid (pB×26) contained a genomic clone of the Adh1 maize gene; the other plasmid (piGUS) was a promoterless construction containing part of intron A of the Adhl gene fused to the gusA coding sequence. Thus, the two vectors shared an effective homologous region consisting of a 459 by (Hindlll—PvuII) fragment of the yAdh1 intron A sequence. An active gusA fusion gene would result upon homologous recombination between the plasmids within the intron A sequence, and indeed GUS activity was observed in extracts following co-transformation of maize protoplasts with the two plasmids. The presence of recombinant DNA molecules in protoplast DNA isolated 1 day after co-transformation was verified using polymerase chain reactions (PCR) and Southern blots. For efficient homologous recombination, both plasmids had to be linearized. The recombination reaction was induced by restriction of the plasmid molecules either inside the effective homologous region or at the borders of the intron sequence. However, the presence of even small, terminal, nonhomologous sequences at the 3′ end of the pB×26 fragment inhibited the recombination reaction. Also, both ends of the linearized piGUS DNA molecules were involved in the recombination reaction. The results revealed some features of homologous recombination reactions occurring in plant cells which cannot be accommodated by mechanisms postulated for similar reactions in animal system and in lower eukaryotes.
    Type of Medium: Electronic Resource
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  • 2
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    In:  CASI
    Publication Date: 2016-06-07
    Description: Various techniques and methods in fluid management are discussed. Propellant on-orbit transfer efficiency from the space shuttle orbiter to orbital transfer vehicle is discussed. Techniques such as refueling, residuals recovery and propellant storage are described.
    Keywords: SPACE TRANSPORTATION
    Type: Satellite Serv. Workshop, Vol. 1; p 381-405
    Format: application/pdf
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  • 3
    Publication Date: 2019-06-27
    Description: Data was obtained on silicone-bonded fiberglass, isocyanurate foam, and two dozen other insulators. Materials were selected to withstand heat sterilization, outer space, and the Martian atmosphere. Significant environmental parameters were vibration, landing shock, and launch venting.
    Keywords: MATERIALS
    Type: NPO-11586
    Format: application/pdf
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