Evaluation of seed storage-protein gene 5' untranslated regions in enhancing gene expression in transgenic rice seed
Evaluation of seed storage-protein gene 5' untranslated regions in enhancing gene expression in transgenic rice seed
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DOI：
10.1007/s00122-010-1386-6
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发表时间：
2010-11-01
影响因子：
5.4
通讯作者：
Qu, Le Qing
中科院分区：
文献类型：
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作者：
Liu, Wen Xian;Liu, Hua Liang;Qu, Le Qing
5'非翻译区 (UTR) 是调节基因表达的重要序列元件。使用由水稻种子贮藏蛋白谷蛋白的 GluC 启动子驱动的 β-葡萄糖醛酸酶 (GUS) 报告基因，我们评估了 6 个种子贮藏蛋白基因的 5'-UTR 在增强稳定转基因水稻品系中外源基因表达水平方面的潜力。所有 5'-UTR 均显着增强 GluC 启动子的表达水平，而不改变其表达模式。 Glb-1和GluA-1的5'-UTR分别使GUS的表达增加约3.36倍和3.11倍。 Glb-1、OsAct2 和 CMV35S 启动子下游的两个 5'-UTR 也增加了稳定转基因水稻品系或瞬时表达原生质体中的 GUS 表达水平。因此，增强与启动子序列无关。实时定量 RT-PCR 分析表明，蛋白质产量的增加并不伴随 mRNA 水平的变化，这表明蛋白质产量的增加是由于 mRNA 翻译效率的提高所致。当与强启动子结合时，Glb-1 和 GluA-1 的 5'-UTR 可能是在水稻种子中高产重组蛋白的理想候选者。
5' untranslated regions (UTRs) are important sequence elements that modulate the expression of genes. Using the beta-glucuronidase (GUS) reporter gene driven by the GluC promoter for the rice-seed storage-protein glutelin, we evaluated the potential of the 5'-UTRs of six seed storage-protein genes in enhancing the expression levels of the foreign gene in stable transgenic rice lines. All of the 5'-UTRs significantly enhanced the expression level of the GluC promoter without altering its expression pattern. The 5'-UTRs of Glb-1 and GluA-1 increased the expression of GUS by about 3.36- and 3.11-fold, respectively. The two 5'-UTRs downstream of the Glb-1, OsAct2 and CMV35S promoters also increased GUS expression level in stable transgenic rice lines or in transient expression protoplasts. Therefore, the enhancements were independent of the promoter sequence. Real-time quantitative RT-PCR analysis showed that the increase in protein production was not accompanied by alteration in mRNA levels, which suggests that the enhancements were due to increasing the translational efficiencies of the mRNA. The 5'-UTRs of Glb-1 and GluA-1, when combined with strong promoters, might be ideal candidates for high production of recombinant proteins in rice seeds.