黄瓜抗坏血酸氧化酶基因5'-上游区域中的伤口响应顺式元件。

Wound-responsive cis-element in the 5'-upstream region of cucumber ascorbate oxidase gene.

作者信息

Asao Hiroshi, Yoshida Kazuya, Nishi Yukio, Shinmyo Atsuhiko

机构信息

Nara Agricultural Experiment Station, Shijo-cho, Kashihara, Nara 634-0813, Japan.

出版信息

Biosci Biotechnol Biochem. 2003 Feb;67(2):271-7. doi: 10.1271/bbb.67.271.

DOI:10.1271/bbb.67.271

PMID:12728985

Abstract

The cucumber (Cucumis sativas) AAO1 gene (former name, Aso1) encodes an ascorbate oxidase that catalyzes the oxidation by molecular oxygen of ascorbic acid to dehydroascorbate. CsAAO1 mRNA concentrations rose rapidly after mechanical wounding of cucumbers. To study the wound-responsive expression of CsAAO1 in detail, we examined transgenic tobacco plants harboring a CsAAO1 promoter-beta-glucuronidase fusion gene. CsAAO1 promoter activity in leaves of the tobacco was induced by wounding. Analysis of the regulatory properties of 5'-deleted promoter fragments showed that a putative wound-responsive cis-element (WRE) was located -736 to -707 bp from the translation initiation site. DNA binding factors that bound specifically to the putative WRE sequence were identified in tobacco nuclear extracts by gel retardation assays.

摘要

黄瓜（Cucumis sativas）AAO1基因（原名Aso1）编码一种抗坏血酸氧化酶，该酶催化分子氧将抗坏血酸氧化为脱氢抗坏血酸。黄瓜受到机械损伤后，CsAAO1 mRNA浓度迅速上升。为了详细研究CsAAO1的伤口响应表达，我们检测了携带CsAAO1启动子- β-葡萄糖醛酸酶融合基因的转基因烟草植株。烟草叶片中CsAAO1启动子活性受伤口诱导。对5'-缺失启动子片段调控特性的分析表明，一个假定的伤口响应顺式元件（WRE）位于翻译起始位点上游-736至-707 bp处。通过凝胶阻滞试验在烟草核提取物中鉴定出与假定WRE序列特异性结合的DNA结合因子。