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Analysis of splice donor and acceptor site function in a transposable gene trap derived from the maize element Activator
Authors:L Nussaume  K Harrison  V Klimyuk  R Martienssen  V Sundaresan and J D G Jones
Institution:(1) Sainsbury Laboratory, John Innes Institute, NR4 7UH Colney Lane, Norwich, UK;(2) Cold Spring Harbor Laboratory, Cold Spring Harbor, P.O. Box 100, 11724 New York, USA;(3) Present address: Laboratoire de Métabolisme Carboné, Centre d'Etude Nucléaire de Cadarache, 13108 St Paul lez Durance Cedex, France
Abstract:Gene trap vectors have been used in insertional mutagenesis in animal systems to clone genes with interesting patterns of expression. These vectors are designed to allow the expression of a reporter gene when the vector inserts into a transcribed region. In this paper we examine alternative splicing events that result in the expression of a GUS reporter gene carried on a Ds element which has been designed as a gene trap vector for plants. We have developed a rapid and reliable method based on PCR to study such events. Many splice donor sites were observed in the 3prime Ac border. The relative frequency of utilisation of certain splice donor and acceptor sites differed between tobacco and Arabidopsis. A higher stringency of splicing was observed in Arabidopsis.
Keywords:Splicing  Transposon tagging  Intron  Ac/Ds
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