Generation of T-DNA Tagging Lines with a Bidirectional Gene Trap Vector and the Establishment of an Insertion-Site Database View Full Text


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Article Info

DATE

2004-03

AUTHORS

Choong-Hwan Ryu, Jung-Hwa You, Hong-Gyu Kang, Junghe Hur, Young-Hea Kim, Min-Jung Han, Kyungsook An, Byoung-Chull Chung, Choon-Hwan Lee, Gynheung An

ABSTRACT

We have developed a binary T-DNA vector, pGA2717, that contains the promoter-less β-glucuronidase (gus) gene adjacent to the right border and the promoter-less green fluorescence protein (gfp) gene next to the left border of the T-DNA. Therefore, inserting T-DNA into a gene can result in the activation of either gus or gfp. A total of 12 169 T-DNA insertional lines of japonica rice were generated using this binary vector. Out of 3140 lines examined, 0.5% of their mature seeds and 2.0% of the 3-day-old etiolated seedlings were GFP-positive. However, GUS assays of the same materials resulted in the identification of 151 (4.8%) GUS-positive lines. Using DNA gel blot and reverse transcription (RT)-PCR analyses, we confirmed that the GFP-positive lines were a true indication of gene trapping. A fusion transcript was also obtained between gfp and the trapped gene. We isolated 990 genomic sequences flanking T-DNA from our analysis of 2099 transgenic plants. Among the insertions, 625 T-DNAs were integrated into genic regions; 361 were located in intergenic regions. These tagging lines will be valuable in trapping and studying various genes for their expression patterns, as well as providing a useful tool for genetic approaches. More... »

PAGES

489-502

References to SciGraph publications

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/b:plan.0000038257.93381.05

    DOI

    http://dx.doi.org/10.1023/b:plan.0000038257.93381.05

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1038436591

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/15316285


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