Department of Biology, Washington University, St Louis, MO 63130, USA.
EMBO J. 1988 Feb;7(2):297-302. doi: 10.1002/j.1460-2075.1988.tb02812.x.
Genes encoding beta-conglycinin, a soybean seed storage protein, are expressed only in seeds during mid-to-late stages in embryogeny. It was previously determined that a DNA sequence 200 nucleotides upstream of the transcriptional start site of the gene encoding the alpha'-subunit of beta-conglycinin is essential for regulated gene expression in transgenic plants. The regulatory effect of this DNA element was tested by inserting the element in different positions and different orientations within a chimeric constitutively expressed reporter gene. The reporter gene was comprised of the 35S promoter from cauliflower mosaic virus (CaMV), a gene encoding chloramphenicol acetyltransferase (CAT) and the polyadenylation signal from the alpha'-subunit gene. The element had no significant effect on the expression of the CAT gene in roots, stems, or leaves, regardless of the position of its insertion (i.e. 5 or 3 of the gene). However, there was 25- to 40-fold enhancement of CAT gene expression in seeds during mid-to-late stages of embryo development when the element was placed in either orientation within the 35S promoter. There was 2- to 4-fold enhancement of CAT activity when the element was placed 3' of the CAT coding sequence. No enhancement was detected when the element was placed downstream of the 3' non-coding region. This is, to our knowledge, the first identification of a cis-acting element that enhances gene expression in a tissue-specific and temporally regulated manner during embryo development in plants.
基因编码β-伴大豆球蛋白,一种大豆种子贮藏蛋白,仅在胚胎发生的中期到后期在种子中表达。先前已经确定,基因编码β-伴大豆球蛋白的α'-亚基的转录起始位点上游的 200 个核苷酸的 DNA 序列对于转基因植物中受调控的基因表达是必需的。通过在嵌合组成型表达报告基因内的不同位置和不同方向插入该元件,测试了该 DNA 元件的调节作用。报告基因由花椰菜花叶病毒(CaMV)的 35S 启动子、编码氯霉素乙酰转移酶(CAT)的基因和α'-亚基基因的多聚腺苷酸化信号组成。无论其插入位置(即基因的 5 或 3')如何,该元件对根、茎或叶中 CAT 基因的表达均没有显著影响。然而,当元件以任一方向置于 35S 启动子内时,在胚胎发育的中期到后期,种子中 CAT 基因的表达增强了 25-40 倍。当元件置于 CAT 编码序列的 3'时,CAT 活性增强了 2-4 倍。当元件置于 3'非编码区的下游时,未检测到增强。这是,据我们所知,首次鉴定出一种顺式作用元件,该元件以组织特异性和胚胎发育过程中时间调控的方式增强植物中基因的表达。