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Rescue of a tk-plasmid from transgenic mice reveals its episomal transmission
Authors:Udo Kiessling   Klaus Becker   Michael Strauss   Joerg Schoeneich  Erhard Geissler
Affiliation:(1) Zentralinstitut für Molekularbiologie der Akademie der Wissenschaften der DDR, Robert-Roessle-Straße 10, DDR-1115 Berlin;(2) Zentralinstitut für Genetik und Kulturpflanzenforschung der Akademie der Wissenschaften der DDR, Corrensstraße 3, DDR-4325 Gatersleben, German Democratic Republic
Abstract:Summary This communication demonstrates the usefulness of the plamid rescue procedure for recovery of plasmids from transgenic mice. We have microinjected the plasmid pSK1 harbouring the Herpes simplex virus thymidine kinase gene into fertilized mouse oocytes and succeeded in recovering plasmids from newborns by transformation of E. coli either with HindIII cut cellular DNA or with uncut DNA. The majority of the rescued plasmids were indistinguishable from pSK1 by restriction analysis. The rescued plasmids proved to be functionally active in a transient expression assay in mouse Ltk- cells. The pSK1 DNA sequences were inherited by up to 90% of the second generation progeny mice, which is not in agreement with a Mendelian transmission of heterozygous markers integrated into a single site of the chromosome.These data support the assumption that germ line transmission of non-integrated episomal plasmids can occur.
Keywords:Transgenic mice  Plasmid rescue  HSV thymidine kinase gene  Extrachromosomal DNA  Transmission
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