Manna F, Massardo D R, Wolf K, Luccarini G, Carlomagno M S, Rivellini F, Alifano P, Del Giudice L
Istituto Internazionale di Genetica e Biofisica, C.N.R., Napoli, Italy.
Nucleic Acids Res. 1994 May 11;22(9):1712-8. doi: 10.1093/nar/22.9.1712.
In vivo analysis of expression of the chloroplast rDNA cluster during somatic embryogenesis of Daucus carota (D.carota) was performed by Northern-blot analysis with different DNA probes, spanning both the 16S rRNA gene, the 16S-23S rRNA spacer, which contains the two mosaic tRNA genes tRNA(Ile) and tRNA(Ala), and the region upstream of the 16S rRNA gene, where a tRNA(Val) maps. We show that expression both of the spacer tRNAs tRNA(Ile) and tRNA(Ala) is not significantly regulated during development whereas the amount of the transcript corresponding to tRNA(Val) is not detectable during early embryonic stages and progressively accumulates during late phases. Multiple transcription start sites have been identified upstream of the tRNA(Val) gene by S1 mapping analysis, which are activated late during the embryogenesis. These data indicate that developmental control mechanisms act on plastid gene expression during embryogenesis in carrot.
利用不同的DNA探针通过Northern杂交分析,对胡萝卜体细胞胚胎发生过程中叶绿体rDNA簇的表达进行了体内分析。这些探针覆盖了16S rRNA基因、16S - 23S rRNA间隔区(其中包含两个镶嵌的tRNA基因tRNA(Ile)和tRNA(Ala))以及16S rRNA基因上游区域(tRNA(Val)定位于此)。我们发现,间隔区tRNA(tRNA(Ile)和tRNA(Ala))的表达在发育过程中没有明显的调控,而对应于tRNA(Val)的转录本在胚胎早期阶段无法检测到,在后期逐渐积累。通过S1图谱分析,在tRNA(Val)基因上游鉴定出多个转录起始位点,这些位点在胚胎发生后期被激活。这些数据表明,发育控制机制在胡萝卜胚胎发生过程中作用于质体基因表达。