Journal Article

Molecular Genetics Using T-DNA in Rice

Gynheung An, Shinyoung Lee, Sung-Hyun Kim and Seong-Ryong Kim

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 46, issue 1, pages 14-22
Published in print January 2005 | ISSN: 0032-0781
Published online January 2005 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci502
Molecular Genetics Using T-DNA in Rice

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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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.

Keywords: Keywords: DNA pools — Functional genomics — Gene tagging — Reverse genetics — Tag end sequence — T-DNA.; Abbreviations: CaMV, cauliflower mosaic virus; GFP, green fluorescent protein; GUS, β-glucuronidase; TAIL PCR, thermal asymmetric interlaced polymerase chain reaction; T-DNA, transferred DNA; TES, tag end sequence.

Journal Article.  7098 words. 

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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