Establishment of gene-trap and enhancer-trap systems in the moss Physcomitrella patens.
Because of its simple body plan and ease of gene knockout and allele replacement, the moss Physcomitrella patens is often used as a model system for studies in plant physiology and developmental biology. Gene-trap and enhancer-trap systems are useful techniques for cloning genes and enhancers that function in specific tissues or cells. Additionally, these systems are convenient for obtaining molecular markers specific for certain developmental processes. Elements for gene-trap and enhancer-trap systems were constructed using the uidA reporter gene with either a splice acceptor or a minimal promoter. Through a high rate of transformation conferred by a method utilizing homologous recombination, 235 gene-trap and 1073 enhancer-trap lines were obtained from 5637 and 3726 transgenic lines, respectively. The expression patterns of these trap lines in the moss gametophyte varied. The candidate gene trapped in a gene-trap line YH209, which shows rhizoid-specific expression, was obtained by 5' and 3' RACE. This gene was named PpGLU, and forms a clade with plant acidic alpha-glucosidase genes. Thus, these gene-trap and enhancer-trap systems should prove useful to identify tissue- and cell-specific genes in Physcomitrella.
Hiwatashi
Y
Y
National Institute for Basic Biology, Okazaki 444-8585, Japan.
Nishiyama
T
T
Fujita
T
T
Hasebe
M
M
D015152:Blotting, Northern; D015139:Blotting, Southern; D019068:Bryopsida / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D018628:Gene Dosage; D018507:Gene Expression Regulation, Developmental; D017343:Genes, Plant / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D010802:Phylogeny; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D005819:Genetic Markers