Hellmund D, Metzlaff M, Serfling E
Nucleic Acids Res. 1984 Nov 12;12(21):8253-68. doi: 10.1093/nar/12.21.8253.
We present the primary structure of a chloroplast tRNAArgACG gene of the plant, Pelargonium zonale, and its faithful expression in Xenopus oocyte nuclei. This tRNAArg gene is located 250 bp downstream of a 5S RNA gene within a cloned 5kb long ribosomal DNA segment (Fig. 1). The Pelargonium tRNAArg gene shares 97% and 86% sequence homology with tRNAArgACG genes of Spirodela oligorhiza and Euglena gracilis chloroplasts, respectively, and also extensive homology (70%) with the corresponding gene of E. coli. It lacks an intervening sequence and, like eukaryotic tRNA genes, does not code for the 3' terminal CCA nucleotides. Moreover, the chloroplast tRNAArg gene carries all the sequence elements essential for transcription by vertebrate RNA polymerase III since it is efficiently expressed in Xenopus oocyte nuclei, even in the presence of 1 microgram/ml alpha-amanitin. In Xenopus oocyte nuclei, no transcripts of the chloroplast 5S RNA gene were detected.
我们展示了天竺葵叶绿体tRNAArgACG基因的一级结构及其在非洲爪蟾卵母细胞核中的稳定表达。该tRNAArg基因位于一个克隆的5kb长核糖体DNA片段内5S RNA基因下游250bp处(图1)。天竺葵tRNAArg基因与少根紫萍和纤细裸藻叶绿体的tRNAArgACG基因的序列同源性分别为97%和86%,与大肠杆菌的相应基因也有广泛的同源性(70%)。它没有内含序列,并且像真核tRNA基因一样,不编码3'末端的CCA核苷酸。此外,叶绿体tRNAArg基因携带了脊椎动物RNA聚合酶III转录所需的所有序列元件,因为它即使在存在1微克/毫升α-鹅膏蕈碱的情况下也能在非洲爪蟾卵母细胞核中高效表达。在非洲爪蟾卵母细胞核中,未检测到叶绿体5S RNA基因的转录本。