Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice
Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice
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DOI：
10.1111/j.1467-7652.2004.00055.x
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发表时间：
2004-03-01
影响因子：
13.8
通讯作者：
Takaiwa, F
中科院分区：
文献类型：
--
作者：
Qu, LQ;Takaiwa, F
利用稳定的转基因水稻植株，分析了水稻种子中表达的15个基因的启动子的时空表达模式及其促进重组蛋白在种子中表达的潜力。这15个基因包括10个种子贮藏蛋白基因和5个碳水化合物和氮代谢相关酶基因。启动子的谷蛋白和13 kDa和16 kDa的醇溶谷蛋白直接胚乳特异性表达，特别是在外部（周边区域）的胚乳，而胚球蛋白和18 kDa的油质蛋白启动子直接在胚和糊粉层的表达。GUS基因与26 kDa球蛋白启动子的融合导致在内部淀粉胚乳组织中表达。应该注意的是，10 kDa醇溶谷蛋白基因是唯一一个需要5'和3'侧翼区进行内源胚乳特异性表达的测试基因。丙酮酸正磷酸二激酶（PPDK）和ADP-葡萄糖焦磷酸化酶（AGP 3）小亚基基因的启动子不仅在种子中有活性，而且在营养组织的韧皮部中也有活性。在种子内，这两种启动子的表达不同之处在于PPDK基因仅在胚乳中表达，而AG β小亚基基因在整个种子中表达。GUS报告基因与丙氨酸氨基转移酶（AlaAT）启动子融合后在淀粉胚乳的内部表达，而淀粉分支酶（SBE 1）和谷氨酸合成酶（GOGAT）基因主要在胚乳和胚之间的盾片中表达。当启动子活性在种子成熟过程中进行了检查，谷蛋白GluB-4，26 kDa球蛋白和10 kDa和16 kDa醇溶蛋白启动子表现出更高的活性比其他。这里分析的种子启动子表现出各种各样的活动和表达模式，从而提供了许多选择，适合于各种应用在植物生物技术。
Using stable transgenic rice plants, the promoters of 15 genes expressed in rice seed were analysed for their spatial and temporal expression pattern and their potential to promote the expression of recombinant proteins in seeds. The 15 genes included 10 seed storage protein genes and five genes for enzymes involved in carbohydrate and nitrogen metabolism. The promoters for the glutelins and the 13 kDa and 16 kDa prolamins directed endospermspecific expression, especially in the outer portion (peripheral region) of the endosperm, whilst the embryo globulin and 18 kDa oleosin promoters directed expression in the embryo and aleurone layer. Fusion of the GUS gene to the 26 kDa globulin promoter resulted in expression in the inner starchy endosperm tissue. It should be noted that the 10 kDa prolamin gene was the only one tested that required both the 5' and 3' flanking regions for intrinsic endosperm-specific expression. The promoters from the pyruvate orthophosphate dikinase (PPDK) and ADP-glucose pyrophosphorylase (AGPase) small subunit genes were active not only in the seed, but also in the phloem of vegetative tissues. Within the seed, the expression from these two promoters differed in that the PPDK gene was only expressed in the endosperm, whereas the AGPase small subunit gene was expressed throughout the seed. The GUS reporter gene fused to the alanine aminotransferase (AlaAT) promoter was expressed in the inner portion of the starchy endosperm, whilst the starch branching enzyme (SBE1) and the glutamate synthase (GOGAT) genes were mainly expressed in the scutellum (between the endosperm and embryo). When promoter activities were examined during seed maturation, the glutelin GluB-4, 26 kDa globulin and 10 kDa and 16 kDa prolamin promoters exhibited much higher activities than the others. The seed promoters analysed here exhibited a wide variety of activities and expression patterns, thus providing many choices suitable for various applications in plant biotechnology.