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Molecular analysis of T-DNA insertion mutant population for transgenic rice plants

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  • URLhttps://db.koreascholar.com/Article/Detail/298464
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한국육종학회 (The Korean Breeding Society)
초록

We have generated 383 independent transgenic lines for genetically modified (GM) rice that contained GPD, UtrCSP, BrTSR15 and BrTSR53 genes overexpression constructs under the control of the constitutive CaMV 35S promoter. TaqMan copy number assay was determined inserted T-DNA copy number. Also FSTs analysis was isolated from 203 single copy T-DNA lines of transgenic plants and sequence mapped to the rice chromosomes. In analyzing single copy lines, we identified 95 FSTs, among which 37 (38.9%) were integrated into genic regions and 58 (61.1%) into intergenic regions. About 27 homozygous lines were obtained through multi-generations of planting, resistance screening and TaqMan copy number assay. To investigate the transgene expression patterns, quantitative real-time PCR analysis was performed using total RNAs from leaf tissue of single copy, intergenic region of T-DNA insertion and homozygous T2 plants. The mRNA expression levels of the examined transgenic rice were significantly increased in all of the transgenic plants. In addition, myc-tagged 35S::BrTSR15 and 35S::BrTSR53 transgenic plants were displayed higher levels of transgene protein than WT plants. These results may be useful for producing of large-scale transgenic plants or T-DNA inserted mutants in rice.

저자
  • Hyemin Lim(National Academy of Agricultural Science, Rural Development Administration)
  • A-Ram Kim(National Academy of Agricultural Science, Rural Development Administration)
  • Jung-Il Cho(National Academy of Agricultural Science, Rural Development Administration)
  • Hyeon-So Ji(National Academy of Agricultural Science, Rural Development Administration)
  • Ung-Han Yoon(National Academy of Agricultural Science, Rural Development Administration)
  • Tae-Ho Kim(National Academy of Agricultural Science, Rural Development Administration)
  • Gang-Seob Lee(National Academy of Agricultural Science, Rural Development Administration) Corresponding Author