Efficient gene targeting by homologous recombination in rice.
Modification of genes through homologous recombination, termed gene targeting, is the most direct method to characterize gene function. In higher plants, however, the method is far from a common practice. Here we describe an efficient and reproducible procedure with a strong positive/negative selection for gene targeting in rice, which feeds more than half of the world's population and is an important model plant. About 1% of selected calli and their regenerated fertile plants were heterozygous at the targeted locus, and only one copy of the selective marker used was found at the targeted site in their genomes. The procedure's applicability to other genes will make it feasible to obtain various gene-targeted lines of rice.
Terada
Rie
R
National Institute for Basic Biology, Okazaki, 444-8585, Japan.
Urawa
Hiroko
H
Inagaki
Yoshishige
Y
Tsugane
Kazuo
K
Iida
Shigeru
S
D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D019020:Directed Molecular Evolution / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D017343:Genes, Plant / Q000235:genetics*; D008969:Molecular Sequence Data; D012275:Oryza / Q000145:classification / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D011786:Quality Control; D011995:Recombination, Genetic; D012231:Rhizobium / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014170:Transformation, Genetic; D019076:Transgenes
D004269:DNA, Bacterial