Maréchal-Drouard L, Guillemaut P, Cosset A, Arbogast M, Weber F, Weil J H, Dietrich A
Institut de Biologie Moléculaire des Plantes du CNRS, Université Louis Pasteur, Strasbourg, France.
Nucleic Acids Res. 1990 Jul 11;18(13):3689-96. doi: 10.1093/nar/18.13.3689.
Total transfer RNAs were extracted from highly purified potato mitochondria. From quantitative measurements, the in vivo tRNA concentration in mitochondria was estimated to be in the range of 60 microM. Total potato mitochondrial tRNAs were fractionated by two-dimensional polyacrylamide gel electrophoresis. Thirty one individual tRNAs, which could read all sense codons, were identified by aminoacylation, sequencing or hybridization to specific oligonucleotides. The tRNA population that we have characterized comprises 15 typically mitochondrial, 5 'chloroplast-like' and 11 nuclear-encoded species. One tRNA(Ala), 2 tRNAs(Arg), 1 tRNA(Ile), 5 tRNAs(Leu) and 2 tRNAs(Thr) were shown to be coded for by nuclear DNA. A second, mitochondrial-encoded, tRNA(Ile) was also found. Five 'chloroplast-like' tRNAs, tRNA(Trp), tRNA(Asn), tRNA(His), tRNA(Ser)(GGA) and tRNA(Met)m, presumably transcribed from promiscuous chloroplast DNA sequences inserted in the mitochondrial genome, were identified, but, in contrast to wheat (1), potato mitochondria do not seem to contain 'chloroplast-like' tRNA(Cys) and tRNA(Phe). The two identified tRNAs(Val), as well as the tRNA(Gly), were found to be coded for by the mitochondrial genome, which again contrasts with the situation in wheat, where the mitochondrial genome apparently contains no tRNA(Val) or tRNA(Gly) gene (2).
从高度纯化的马铃薯线粒体中提取了总转运RNA。通过定量测量,估计线粒体中体内tRNA浓度在60微摩尔范围内。用二维聚丙烯酰胺凝胶电泳对马铃薯线粒体总tRNA进行分级分离。通过氨酰化、测序或与特定寡核苷酸杂交鉴定出31种能识别所有有义密码子的单个tRNA。我们所鉴定的tRNA群体包括15种典型的线粒体tRNA、5种“类叶绿体”tRNA和11种核编码tRNA。已证明1种tRNA(Ala)、2种tRNA(Arg)、1种tRNA(Ile)、5种tRNA(Leu)和2种tRNA(Thr)由核DNA编码。还发现了第二种由线粒体编码的tRNA(Ile)。鉴定出5种“类叶绿体”tRNA,即tRNA(Trp)、tRNA(Asn)、tRNA(His)、tRNA(Ser)(GGA)和tRNA(Met)m,推测它们是从插入线粒体基因组的混杂叶绿体DNA序列转录而来的,但与小麦不同(1),马铃薯线粒体似乎不包含“类叶绿体”tRNA(Cys)和tRNA(Phe)。已鉴定的2种tRNA(Val)以及tRNA(Gly)被发现由线粒体基因组编码,这再次与小麦的情况形成对比,在小麦中,线粒体基因组显然不包含tRNA(Val)或tRNA(Gly)基因(2)。