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Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
An expression vector inhibits gene expression in Xenopus embryos by antisense RNA
Michael Schmid1, Herbert Steinbeisser2, Hans-Henning Epperlein3
1Medizinisch-Naturwissenschaftliches Forschungszentrum, Universität Tübingen, Ob dem Himmelreich 7, W-7400, Tübingen, Germany.
Abstract:
An expression vector was constructed containing the entire bovine papilloma virus (BPV-1) genome and part of the a-actin gene of Xenopus laevis cloned in the antisense orientation into the neomycin resistance gene under the control of the herpes simplex virus (HSV) thymidine kinase (TK) promoter. When this vector is microinjected into X. laevis embryos it replicates extrachromosomally, at least up to the tadpole stage, and a fusion RNA is synthesized after the mid blastula transition (MBT). The expression of the antisense gene results in a morphological abnormality of somites demonstrating that antisense RNA generated by an episomal replicating expression vector can inhibit the expression of a selected gene during early embryogenesis of X. laevis.
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