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植物细胞核中的tRNA(甲硫氨酸)基因普遍被内含子打断。

Plant nuclear tRNA(Met) genes are ubiquitously interrupted by introns.

作者信息

Akama K, Kashihara M

机构信息

Department of Biology, Faculty of Science, Shimane University, Matsue, Japan.

出版信息

Plant Mol Biol. 1996 Nov;32(3):427-34. doi: 10.1007/BF00019094.

Abstract

We have isolated three independent clones for nuclear elongator tRNA(Met) genes from an Arabidopsis DNA library using a tRNA(Met)-specific probe generated by PCR. Each of the coding sequences for tRNAMet in these clones is identical and is interrupted by an identical 11 bp long intervening sequence at the same position in the anticodon loop of the tRNA. Their sequences differ at two positions from the intron in a soybean counterpart. Southern analysis of Arabidopsis DNA demonstrates that a gene family coding for tRNA(Met) is dispersed at at least eight loci in the genome. The unspliced precursor tRNA(Met) intermediate was detected by RNA analysis using an oligonucleotide probe complementary to the putative intron sequence. In order to know whether introns commonly interrupt plant tRNA(Met) genes, their coding sequences were PCR-amplified from the DNAs of eight phylogenetically separate plant species. All 53 sequences determined contain 10 to 13 bp long intervening sequences, always positioned one base downstream from the anticodon. They can all be potentially folded into the secondary structure characteristic for plant intron-containing precursor tRNAs. Surprisingly, GC residues are always present at the 5'-distal end of each intron.

摘要

我们使用通过PCR产生的tRNA(Met)特异性探针,从拟南芥DNA文库中分离出了三个独立的核延长因子tRNA(Met)基因克隆。这些克隆中每个tRNAMet的编码序列都是相同的,并且在tRNA反密码子环的相同位置被一个相同的11 bp长的间隔序列中断。它们的序列与大豆对应物中的内含子在两个位置上不同。对拟南芥DNA的Southern分析表明,编码tRNA(Met)的基因家族分散在基因组的至少八个位点。使用与推定内含子序列互补的寡核苷酸探针通过RNA分析检测到未剪接的前体tRNA(Met)中间体。为了了解内含子是否普遍中断植物tRNA(Met)基因,从八个系统发育上不同的植物物种DNA中PCR扩增了它们的编码序列。所确定的所有53个序列都包含10到13 bp长的间隔序列,总是位于反密码子下游一个碱基处。它们都可以潜在地折叠成含有植物内含子的前体tRNA的二级结构特征。令人惊讶的是,GC残基总是存在于每个内含子的5'远端。

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