Lindstrom J T, Vodkin L O, Harding R W, Goeken R M
Department of Agronomy, University of Illinois, Urbana.
Dev Genet. 1990;11(2):160-7. doi: 10.1002/dvg.1020110206.
A series of constructs containing the developmentally regulated soybean lectin gene (Le1) were used to transform tobacco plants in order to assess developmental and quantitative regulation conferred by flanking sequences. The largest of the lectin constructs contained approximately 3,000 base pairs (bp) of Le1 5 flanking region and 1,500 bp of the 3 flanking region. The smallest construct contained no 5 flanking region and 194 bp of the 3 flanking region. ELISA assays of lectin in individual tobacco seeds and Southern blot analyses confirmed that most constructs were inherited as unique insertion events. Maximal expression of Le1 required more than 338 bp of 5 sequence, indicating that far upstream factors are involved in quantitative control of lectin expression. Lectin expression declined more than 80% between deletions with 1,700 versus 338 bp of 5 flanking sequence. In contrast, developmental control of lectin expression was maintained by Le1 inserts with only 190 bp of 5 sequence. The lectin promoter offers a potential means to target high levels of gene expression to the developing seeds of soybean or other dicotyledonous plants.
为了评估侧翼序列赋予的发育调控和定量调控,使用了一系列包含发育调控的大豆凝集素基因(Le1)的构建体来转化烟草植株。最大的凝集素构建体包含约3000个碱基对(bp)的Le1 5'侧翼区域和1500 bp的3'侧翼区域。最小的构建体不包含5'侧翼区域,仅包含194 bp的3'侧翼区域。对单个烟草种子中的凝集素进行ELISA分析和Southern印迹分析证实,大多数构建体作为独特的插入事件遗传。Le1的最大表达需要超过338 bp的5'序列,这表明远上游因子参与凝集素表达的定量控制。在具有1700 bp与338 bp的5'侧翼序列的缺失之间,凝集素表达下降超过80%。相比之下,仅具有190 bp的5'序列的Le1插入片段维持了凝集素表达的发育控制。凝集素启动子提供了一种潜在的方法,可将高水平的基因表达靶向大豆或其他双子叶植物的发育种子。