Molecular genetics using T-DNA in rice.
Now that sequencing of the rice genome is nearly completed, functional analysis of its large number of genes is the next challenge. Because rice is easy to transform, T-DNA has been used successfully to generate insertional mutant lines. Collectively, several laboratories throughout the world have established at least 200,000 T-DNA insertional lines. Some of those carry the GUS or GFP reporters for either gene or enhancer traps. Others are activation tagging lines for gain-of-function mutagenesis when T-DNA is inserted in the intergenic region. A forward genetic approach showed limited success because of somaclonal variations induced during tissue culture. To utilize these resources more efficiently, tagged lines have been produced for reverse genetics approaches. DNA pools of the T-DNA-tagged lines have been prepared for polymerase chain reaction (PCR) screening of insertional mutants in a given gene. Appropriate T-DNA insertion sites are determined by sequencing the region flanking the T-DNA. This information is then used to make databases that are shared with the scientific community. International efforts on seed amplification and maintenance are needed to exploit these valuable materials efficiently.
An
Gynheung
G
National Research Laboratory of Plant Functional Genomics, Division of Molecular and Life Sciences, Pohang University of Science and Technology (POSTECH), Pohang 790-784, Korea. genean@postech.ac.kr
Lee
Shinyoung
S
Kim
Sung-Hyun
SH
Kim
Seong-Ryong
SR
D004269:DNA, Bacterial / Q000235:genetics*; D030561:Databases, Nucleic Acid; D015723:Gene Library; D023281:Genomics; D008967:Molecular Biology; D016254:Mutagenesis, Insertional; D012275:Oryza / Q000235:genetics* / Q000382:microbiology
D004269:DNA, Bacterial; C036483:T-DNA