Park Hyeong Cheol, Kim Man Lyang, Kang Yun Hwan, Jeong Jae Cheol, Cheong Mi Sun, Choi Wonkyun, Lee Sang Yeol, Cho Moo Je, Kim Min Chul, Chung Woo Sik, Yun Dae-Jin
Division of Applied Life Science, Plant Molecular Biology and Biotechnology Research Center and Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju, Korea.
Mol Cells. 2009 Apr 30;27(4):475-80. doi: 10.1007/s10059-009-0063-6. Epub 2009 Apr 13.
The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.
大豆(Glycine max)钙调蛋白亚型4(GmCaM - 4)的转录在暴露于病原体或NaCl后0.5小时内会显著诱导。先前已在GmCaM - 4启动子中鉴定出响应病原体和盐胁迫调节GmCaM - 4基因表达的核心顺式作用元件,分别位于-1207至-1128 bp以及-858至-728 bp之间。在此,我们表征了在病原体处理的核提取物中,在GmCaM - 4启动子的两个核心顺式作用元件处形成的DNA结合复合物的特性。我们构建了含有约1.3 kb GmCaM - 4启动子各种缺失的GUS报告基因构建体,并分析了用这些构建体转化的烟草植株中的GUS表达。GUS表达分析表明,先前鉴定的两个核心区域参与了在异源系统中诱导GmCaM - 4表达。最后,拟南芥原生质体的瞬时表达分析表明,在病原体或NaCl处理后,GmCaM - 4启动子产生的GUS活性水平高于CaMV35S启动子,这表明GmCaM - 4启动子可能有助于构建条件基因表达系统。