A general method to isolate genes tagged by a high copy number transposable element.
The Petunia hybrida line W138 contains more than 200 copies of the transposable element dTph1. In W138 progeny these elements give rise to new unstable mutations at high frequency. With the aim of isolating these mutated genes a method was developed to isolate dTph1 flanking sequences unique for mutant plants. This method is based on differential screening of cloned inverse polymerase chain reaction (IPCR) products originating from the mutated plant. It directly yields a probe for the mutated gene which can be used to screen pre-existing cDNA and genomic libraries. This method may be generally applicable to isolate genes tagged by other high copy number transposable elements, like Mutator (Mu) or Dissociation (Ds) in Zea mays.
Souer
E
E
Department of Genetics, Vrije Universiteit, Amsterdam, The Netherlands.
Quattrocchio
F
F
de Vetten
N
N
Mol
J
J
Koes
R
R
D000483:Alleles; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004251:DNA Transposable Elements; D018744:DNA, Plant / Q000235:genetics; D005784:Gene Amplification; D017343:Genes, Plant; D005821:Genetic Techniques; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009154:Mutation; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction; D016324:Sequence Tagged Sites
D017931:DNA Primers; D004251:DNA Transposable Elements; D018744:DNA, Plant