Jasmonate-inducible expression of a potato cathepsin D inhibitor-GUS gene fusion in tobacco cells
Jasmonate-inducible expression of a potato cathepsin D inhibitor-GUS gene fusion in tobacco cells
复制标题
茉莉酸诱导的马铃薯组织蛋白酶 D 抑制剂-GUS 基因融合在烟草细胞中的表达
作者：
A. Ishikawa;T. Yoshihara;K. Nakamura
编码组织蛋白酶D抑制剂（CDI）的马铃薯基因在块茎和花蕾中组成型表达，并且在组织受伤或通过用茉莉酸甲酯（MJA）处理时，其在叶中是可诱导的。将CDI基因的2.4kb启动子与β-葡萄糖醛酸苷酶（GUS）编码序列融合，得到的融合基因（CDI：GUS）在烟草悬浮转化细胞中可被MJA诱导表达。在生长素和生长素的MJA诱导的融合基因的表达的抑制的情况下获得的诱导的最大水平被释放的aphidicolin，结果表明，MJA诱导的表达在活跃的细胞分裂过程中被抑制。JA和MJA在诱导融合基因表达方面表现出相似的活性，而其他JA相关化合物如葫芦酸、块茎酸和二氢茉莉酸既不诱导融合基因的表达，也不抑制MJA诱导的融合基因的表达。二氢茉莉酸甲酯特异性地刺激MJA诱导的融合基因的表达。即使用100 bp长的CDI基因启动子也观察到融合基因的MJA诱导型表达，尽管与2.4 kb长的启动子相比表达水平显著降低。100 bp长的CDI启动子不含G-box或六聚体基序，这些基序与其他几种植物基因的MJA响应性表达有关。通过缺失或寡核苷酸插入对100 bp长启动子的进一步诱变表明，尽管-100和-82之间的序列是MJA响应性表达所需的，但单独存在该序列并不赋予MJA响应性表达。
A potato gene encoding cathepsin D inhibitor (CDI) is expressed constitutively in tubers and flower buds and it is inducible in leaves upon wounding of the tissue or by treatment with methyl jasmonate (MJA). A fusion gene (CDI:GUS) in which the 2.4 kb long promoter of the CDI gene was translationaly fused with the coding sequence for β-glucuronidase (GUS) showed MJA-inducible expression in transformed tobacco cells in suspension. The maximum level of induction by MJA was obtained in the absence of auxin and repression of MJA-inducible expression of the fusion gene by auxin was released by aphidicolin, the results suggesting that MJA-inducible expression is repressed during active cell division. JA and MJA showed similar activities in inducing the expression of the fusion gene, while other JA-related compounds such as cucurbic acid, tuberonic acid and dihydrojasmonic acid neither induced expression of the fusion gene nor inhibited the MJA-inducible expression of the fusion gene. Methyl dihydrojasmonate specifically stimulated the MJA-inducible expression of the fusion gene. The MJA-inducible expression of the fusion gene was observed even with a 100 bp long promoter of the CDI gene albeit with significantly decreased level of expression compared to the 2.4 kb long promoter. The 100 bp long CDI promoter did not contain a G-box or hexamer motif that had been implicated in the MJA-responsive expression of several other plant genes. Further mutagenesis of the 100 bp long promoter by deletion or oligonucleotide insertion suggested that although a sequence between −100 and −82 is required for the MJA-responsive expression, the presence of this sequence alone does not confer the MJA-responsive expression.
影响因子：
5.6
作者：
IZAWA, T;FOSTER, R;CHUA, NH
通讯作者：
CHUA, NH