title: Selecting karyophilic DNA cis elements in Xenopus laevis oocytes; a new approach.
abstract: The intracellular localisation and mobility of exogenous DNA introduced into Xenopus laevis oocytes is largely unknown. In this paper, we report a new technique to investigate the cytoplasmic/nuclear transport of a random pool of linear, double-stranded, oligomeric DNA of 147 bp in length. We chose a combinatorial approach which made use of repetitive rounds of selection and amplification to search for new cis elements mediating nuclear import or retention. A new PCR-based methodology was established to reliably detect exogenous DNA in subcellular and total extracts prepared from Xenopus laevis oocytes. Studies in vivo and with cellular extracts indicate the presence of a highly efficient nuclease activity in the nuclear compartment. The described combinatorial approach constitutes a promising tool for the isolation of novel DNA cis elements which may play an important role in the nuclear internalisation and retention of exogenous DNA in Xenopus laevis oocytes.
authors.affiliation: Deutsches Krebsforschungszentrum, Heidelberg, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Marcello
authors.forename: Roger
authors.initials: R
authors.lastname: Fischer
authors.forename: Helmut
authors.initials: H
authors.lastname: Tröster
authors.forename: Michael F
authors.initials: MF
authors.lastname: Trendelenburg
authors.forename: Georg
authors.initials: G
authors.lastname: Sczakiel
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002467:Cell Nucleus / Q000235:genetics; D003593:Cytoplasm / Q000235:genetics; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D005260:Female; D009865:Oocytes / Q000502:physiology*; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D014982:Xenopus laevis / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004247:DNA