Introns and their positions affect the translational activity of mRNA in plant cells
Introns and their positions affect the translational activity of mRNA in plant cells
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DOI：
10.1093/embo-reports/kve090
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发表时间：
2001-05-01
期刊：
影响因子：
7.7
通讯作者：
Lonsdale, DM
中科院分区：
文献类型：
--
作者：
Bourdon, V;Harvey, A;Lonsdale, DM
为了进一步增加植物中的转基因表达水平超过用5'非翻译前导内含子获得的增强，将三个不同的玉米内含子插入荧光素酶报告基因的编码序列内的三个不同位置。将构建体转化到玉米（Black Mexican Sweet）细胞和原生质体中，并测定它们的活性。虽然所有测试的内含子都被正确剪接，但只有其中一个位于特定位置的内含子能够增强基因表达。使用正确的剪接位点去除内含子，产生的荧光素酶mRNA的量没有显着差异。这些数据表明，内含子的位置和序列对表达水平具有显著影响，表明前体mRNA的核加工通过mRNP的结构决定最终表达水平。
In an attempt to further increase transgene expression levels in plants over and above the enhancement obtained with a 5' untranslated leader intron, three different maize introns were inserted at three different positions within the coding sequence of the luciferase reporter gene. Constructs were transformed into maize (Black Mexican Sweet) cells and protoplasts, and their activity determined. Although all introns tested were correctly spliced, only one of them in a particular position was able to enhance gene expression. Correct splicing sites were used for intron removal and the quantity of luciferase mRNA produced did not differ significantly. These data indicate that both the position and the sequence of an intron have marked effects on expression levels, suggesting that nuclear processing of the pre-mRNA determines final expression levels through the structure of the mRNP.