Evaluation of seed storage protein gene 3′-untranslated regions in enhancing gene expression in transgenic rice seed
Evaluation of seed storage protein gene 3′-untranslated regions in enhancing gene expression in transgenic rice seed
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DOI：
10.1007/s11248-011-9552-4
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发表时间：
2012-06-01
影响因子：
3
通讯作者：
Qu, Le Qing
中科院分区：
文献类型：
--
作者：
Li, Wen Jing;Dai, Ling Ling;Qu, Le Qing
3′非翻译区（utr）是调控基因表达的重要序列元件。我们评估了9个种子储存蛋白（SSP）基因的3'- utr作为终止子在稳定的转基因水稻品系中促进由谷氨酸启动子驱动的-葡萄糖醛酸酶（）报告基因表达的潜力。其中6个3'- utr显著提高了-启动子的活性，但没有改变其组织特异性，但改变了其在胚乳中的表达模式。-、-和3'- utr的表达量分别比Nos终止物高3.12倍、2.45倍和2.14倍。这3个3’- utr与启动子、-和启动子结合后，在稳定的转基因水稻品系和原生质体中瞬时表达的GUS水平也有所提高，这表明GUS水平的提高与启动子序列无关。蛋白质产量的增加伴随着mRNA水平的改变，这表明这种增强是由于转录物水平的增加。-、-和的3'- utr与强启动子结合，可能是水稻种子中高产重组蛋白的理想候选者。在单基因或多基因转化中，9个ssp3 '- utr可以作为忠实的终止子，避免同源性基因沉默。
3' untranslated regions (UTRs) are important sequence elements that modulate the expression of genes. We evaluated the potential of the 3'-UTRs of 9 seed storage protein (SSP) genes as terminators in enhancing the expression of the -glucuronidase () reporter gene driven by the glutelin - promoter in stable transgenic rice lines. Six of the 3'-UTRs significantly enhanced the activity of the - promoter without changing its tissue specificity but altered its expression pattern in endosperm. With the 3'-UTRs of -, - and , the expression of was higher by 3.12-, 2.45- and 2.14-fold, respectively, than with the Nos terminator. These three 3'-UTRs, combined with , - and promoters, also increased GUS levels in stable transgenic rice lines or in transient expression in protoplasts, which indicated that the enhancements were independent of the promoter sequence. The increase in protein production was accompanied by altered mRNA levels, which suggests that the enhancements were due to increased transcript level. The 3'-UTRs of -, - and , when combined with strong promoters, might be ideal candidates for high production of recombinant proteins in rice seeds. The 9 SSP 3'-UTRs could function as faithful terminators in mono- or multi-gene transformation avoiding homology-based gene silencing.