Cock J M, Swarup R, Dumas C
Reproduction et Développement des Plantes, UMR 9938 CNRS-INRA-ENSL, Ecole Normale Supérieure de Lyon, France.
Mol Gen Genet. 1997 Aug;255(5):514-24. doi: 10.1007/s004380050524.
Gene expression can be inhibited by antisense RNA transcripts. Although this phenomenon is widely used to analyse gene function in plants, the molecular mechanisms involved are poorly understood. One approach to improving our understanding of antisense gene regulation is to analyse the function of endogenous antisense transcripts. To date, only a small number of plant genes have been shown to be transcribed in both directions and limited information is available concerning the role of natural antisense transcripts in plants. In this study, we have identified several natural antisense transcripts which hybridise to probes derived from the S locus receptor kinase gene (SRK). The RNase protection assay and reverse trancriptase-PCR were used to demonstrate that a proportion of the antisense transcripts are encoded directly by SRK. Using different RNase protection probes, regions of the promoter, exon I (which encodes the S domain) and intron I of SRK were shown to be transcribed in an antisense direction. An antisense SRK transcript was shown to inhibit translation of a sense transcript in vitro. The possible role of antisense SRK transcripts in vivo is discussed.
基因表达可被反义RNA转录物抑制。尽管这一现象在植物基因功能分析中被广泛应用,但其涉及的分子机制仍知之甚少。增进我们对反义基因调控理解的一种方法是分析内源性反义转录物的功能。迄今为止,仅有少数植物基因被证明可双向转录,关于天然反义转录物在植物中的作用的信息也很有限。在本研究中,我们鉴定出了几种与来源于S位点受体激酶基因(SRK)的探针杂交的天然反义转录物。核糖核酸酶保护试验和逆转录聚合酶链反应被用于证明一部分反义转录物直接由SRK编码。使用不同的核糖核酸酶保护探针,SRK的启动子、外显子I(编码S结构域)和内含子I区域被证明以反义方向转录。一种反义SRK转录物在体外被证明可抑制正义转录物的翻译。本文讨论了反义SRK转录物在体内可能的作用。