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一个突变的凝集素基因从一个阻断其表达的插入元件中拯救出来。

A mutant lectin gene is rescued from an insertion element that blocks its expression.

作者信息

Okamuro J K, Goldberg R B

机构信息

Department of Biology, University of California, Los Angeles 90024-1606.

出版信息

Plant Cell. 1992 Sep;4(9):1141-6. doi: 10.1105/tpc.4.9.1141.

Abstract

The soybean lectin gene Le1 encodes a prevalent seed protein and is highly regulated during the life cycle. The mutant lectin gene allele le1 is not transcribed detectably, contains a 3.5-kb Tgm1 insertion element within its coding region 0.6 kb 3' to the transcription start site, and leads to a lectinless phenotype. To determine whether the Tgm1 element or a secondary mutation was responsible for repressing le1 gene transcription, we eliminated the insertion element by constructing a chimeric lectin gene (le1/Le1) that contained the 5' half of the le1 gene and its promoter region and the 3' half of the wild-type Le1 gene. Transformed tobacco seed containing the le1/Le1 gene produced both lectin mRNA and protein, demonstrating that the mutant lectin gene control region is transcriptionally competent. By contrast, transformed seed containing the le1 gene produced no detectable lectin mRNA. We conclude that the absence of detectable transcription from the le1 gene is due to transcriptional inhibition by the Tgm1 insertion element and that this element acts at a distance to block transcription from an upstream promoter region.

摘要

大豆凝集素基因Le1编码一种普遍存在的种子蛋白,并且在生命周期中受到高度调控。突变的凝集素基因等位基因le1无法检测到转录,在其转录起始位点下游0.6 kb的编码区域内含有一个3.5 kb的Tgm1插入元件,并导致无凝集素表型。为了确定是Tgm1元件还是二次突变导致le1基因转录受到抑制,我们通过构建一个嵌合凝集素基因(le1/Le1)来消除插入元件,该基因包含le1基因的5'半部分及其启动子区域以及野生型Le1基因的3'半部分。含有le1/Le1基因的转基因烟草种子产生了凝集素mRNA和蛋白质,表明突变凝集素基因的控制区域具有转录活性。相比之下,含有le1基因的转基因种子未产生可检测到的凝集素mRNA。我们得出结论,le1基因无法检测到转录是由于Tgm1插入元件的转录抑制作用,并且该元件在远距离起作用以阻断上游启动子区域的转录。

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