Dong Qing, Jiang Haiyang, Xu QianQian, Li Xiaoming, Peng Xiaojian, Yu Haibing, Xiang Yan, Cheng Beijiu
Key Lab of Biomass Improvement and Conversion, Anhui Agricultural University, Hefei, 230036, China.
Appl Biochem Biotechnol. 2015 Feb;175(3):1344-57. doi: 10.1007/s12010-014-1277-4. Epub 2014 Nov 13.
The use of tissue-specific promoters to drive the expression of target genes during certain developmental stages or in specific organs can prevent unnecessary gene expression caused by constitutive promoters. Utilizing heterologous promoters to regulate the expression of genes in transgenic receptors can help prevent gene silencing. Here, we engineered heterologous maize promoters that regulate gene-specific expression in rice plant receptors. We performed a histochemical and quantitative β-glucuronidase (GUS) analysis of the Zea mays legumin1 (ZM-LEGF) gene promoter and detailed detection of stably transformed rice expressing the GUS gene under the control of the promoter of ZM-LEGF (pZM-LEGF) and its truncated promoters throughout development. When the promoter sequence was truncated, the location and intensity of GUS expression changed. The results suggest that the sequence from -140 to +41 is a critical region that confers the expression of the entire promoter. Truncation of pZM-LEG (3'-deleted region of pZM-LEGF) markedly increased the GUS activity, with the core cis-elements located in the -273 to -140 regions, namely pZM-LEG6. Detailed analysis of pZM-LEG6::GUS T2 transformant rice seeds and plant tissues at different developmental stages indicated that this promoter is an ideal vegetative tissue-specific promoter that can serve as a valuable tool for transgenic rice breeding and genetic engineering studies.
利用组织特异性启动子在特定发育阶段或特定器官中驱动靶基因的表达,可以防止组成型启动子引起的不必要的基因表达。利用异源启动子调控转基因受体中基因的表达有助于防止基因沉默。在此,我们构建了可调控水稻受体中基因特异性表达的异源玉米启动子。我们对玉米豆球蛋白1(ZM-LEGF)基因启动子进行了组织化学和定量β-葡萄糖醛酸酶(GUS)分析,并在整个发育过程中对在ZM-LEGF(pZM-LEGF)启动子及其截短启动子控制下稳定表达GUS基因的水稻进行了详细检测。当启动子序列被截短时,GUS表达的位置和强度发生了变化。结果表明,从-140到+41的序列是赋予整个启动子表达的关键区域。pZM-LEG(pZM-LEGF的3'缺失区域)的截短显著增加了GUS活性,核心顺式元件位于-273至-140区域,即pZM-LEG6。对不同发育阶段的pZM-LEG6::GUS T2转化体水稻种子和植物组织的详细分析表明,该启动子是一种理想的营养组织特异性启动子,可作为转基因水稻育种和基因工程研究的宝贵工具。