Abstract
This paper reports our attempts to characterize transgene integration sites in transgenic mouse lines generated by the microinjection of large (from 30 to 145 kb) pig DNA fragments encompassing a mammary specific gene, the whey acidic protein gene (WAP). Among the various methods used, the thermal asymmetric interlaced (TAIL-) PCR method allowed us (1) to analyze transgene/genomic borders and internal concatamer junctions for eleven transgenic lines, (2) to obtain sequence information for seven borders, (3) to place three transgenes in the mouse genome, and (4) to obtain sequence data for seven transgene junctions in concatamers. Finally, we characterized various rearrangements in the borders and the inner parts of the transgene. The possibility of such complex rearrangements should be carefully considered when transgenic animals are produced with large genomic DNA fragments.
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Aknowledgments
This work was supported by the ANR program ANR-06-POGM-008_TRANSINTEX. We thank P. Ratet (Institut des Sciences du Végétal, CNRS, Gif-sur-Yvette, France) for his fruitful advices concerning the use of TAIL-PCR method and the members of the mouse facility (UEAR, Jouy-en-Josas, France) for the maintenance of animals.
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Le Saux, A., Houdebine, LM. & Jolivet, G. Chromosome integration of BAC (bacterial artificial chromosome): evidence of multiple rearrangements. Transgenic Res 19, 923–931 (2010). https://doi.org/10.1007/s11248-010-9368-7
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DOI: https://doi.org/10.1007/s11248-010-9368-7