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Identification of a promoter sequence from the betl1 gene cluster able to confer transfer-cell-specific expression in transgenic maize

机译:从betl1基因簇中鉴定能够在转基因玉米中赋予转移细胞特异性表达的启动子序列

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The maize (Zea mays L.) betl1 locus, encoding a basal endosperm transfer layer-specific protein, has been mapped and molecularly cloned in its entirety. The locus is shown to consist of three gene copies in the maize inbred line A69Y. To distinguish the three transcription units from the locus name, we have termed them BETL1a, BETL1b, and BETL1c. Two of the copies are expressed, whereas one is inactive and contains retrotransposon-like insertions in both promoter and intron regions. Based on this information, and a restriction site map covering 17 kb around the BETL1 locus, a DNA fragment putatively containing an active promoter sequence was identified. This fragment was tested for its ability to confer transfer-cell-specific expression in transient and stably transformed maize tissues. The transgenic maize plants obtained showed the predicted cell-type specificity of expression restricted to the basal endosperm transfer cells, although there were minor deviations in promoter strength and timing and accumulation of the transgene product from the corresponding BETL-1 endogene expression pattern.
机译:编码基础胚乳转移层特异性蛋白的玉米(Zea mays L.)betl1基因座已作图,并进行了分子克隆。在玉米自交系A69Y中显示基因座由三个基因拷贝组成。为了将三个转录单位与基因座名称区分开,我们将它们称为BETL1a,BETL1b和BETL1c。表达了两个拷贝,而一个拷贝是无活性的,并且在启动子和内含子区域均包含逆转座子样插入。基于该信息,以及在BETL1基因座周围覆盖17 kb的限制性酶切位点图,鉴定了推测含有活性启动子序列的DNA片段。测试了该片段在瞬时和稳定转化的玉米组织中赋予转移细胞特异性表达的能力。所获得的转基因玉米植物显示出预测的细胞类型表达仅限于基础胚乳转移细胞,尽管启动子强度和转基因产物的启动子强度和时间与积累与相应的BETL-1内生基因表达模式略有差异。

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