Analysis of splice donor and acceptor site function in a transposable gene trap derived from the maize element Activator.
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 3' 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.
Nussaume
L
L
Sainsbury Laboratory, John Innes Institute, Norwich, UK.
Harrison
K
K
Klimyuk
V
V
Martienssen
R
R
Sundaresan
V
V
Jones
J D
JD
D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017931:DNA Primers; D004251:DNA Transposable Elements; D018076:DNA, Complementary / Q000096:biosynthesis; D017343:Genes, Plant; D005966:Glucuronidase / Q000096:biosynthesis; D007438:Introns; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D018749:RNA, Plant / Q000032:analysis / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D013045:Species Specificity; D014026:Nicotiana / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D003313:Zea mays / Q000235:genetics*
D017931:DNA Primers; D004251:DNA Transposable Elements; D018076:DNA, Complementary; D012333:RNA, Messenger; D018749:RNA, Plant; D011994:Recombinant Proteins; D005966:Glucuronidase