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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Telomere-mediated chromosomal truncation in maize
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Telomere-mediated chromosomal truncation in maize

机译：端粒介导的玉米染色体截短

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摘要

Direct repeats of Arabidopsis telomeric sequence were constructed to test telomere-mediated chromosomal truncation in maize. Two constructs with 2.6 kb of telomeric sequence were used to transform maize immature embryos by Agrobacterium-mediated transformation. One hundred seventy-six transgenic lines were recovered in which 231 transgene loci were revealed by a FISH analysis. To analyze chromosomal truncations that result in transgenes located near chromosomal termini, Southern hybridization analyses were performed. A pattern of smear in truncated lines was seen as compared with discrete bands for internal integrations, because telomeres in different cells are elongated differently by telomerase. When multiple restriction enzymes were Used to map the transgene positions, the size of the smears shifted in accordance with the locations of restriction sites on the construct. This result demonstrated that the transgene was present at the end of the chromosome immediately before the integrated telomere sequence. Direct evidence for chromosomal truncation came from the results of FISH karyotyping, which revealed broken chromosomes with transgene signals at the ends. These results demonstrate that telomere-mediated chromosomal truncation operates in plant species. This technology will be useful for chromosomal engineering in maize as well as other plant species.
机译：构建了拟南芥端粒序列的直接重复序列，以测试玉米中端粒介导的染色体截短。通过土壤杆菌介导的转化，使用两个具有2.6 kb端粒序列的构建体来转化玉米未成熟胚。回收了176个转基因品系，其中通过FISH分析揭示了231个转基因基因座。为了分析导致转基因位于染色体末端附近的染色体截短，进行了Southern杂交分析。与用于内部整合的离散条带相比，可以看到截断线的涂片模式，因为不同细胞中的端粒被端粒酶不同地延长。当使用多种限制酶对转基因位置作图时，涂片的大小根据构建体上限制位点的位置而变化。该结果表明转基因存在于染色体的末端，紧接在整合的端粒序列之前。染色体截短的直接证据来自FISH核型分析的结果，该结果揭示了末端带有转基因信号的破碎染色体。这些结果证明端粒介导的染色体截短在植物物种中起作用。该技术对于玉米以及其他植物物种的染色体工程将很有用。

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