首页> 美国卫生研究院文献>Nucleic Acids Research >Improved methods for structure probing in large RNAs: a rapid heterologous sequencing approach is coupled to the direct mapping of nuclease accessible sites. Application to the 5 terminal domain of eukaryotic 28S rRNA.

【2h】

Improved methods for structure probing in large RNAs: a rapid heterologous sequencing approach is coupled to the direct mapping of nuclease accessible sites. Application to the 5 terminal domain of eukaryotic 28S rRNA.

机译：改进的在大型RNA中探测结构的方法：快速的异源测序方法与核酸酶可及位点的直接作图相结合。应用于真核28S rRNA的5末端结构域。

代理获取

本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。

页面导航

摘要
著录项
相似文献
相关主题

摘要

We have developed a combined approach for probing native structures in large RNAs. In the first method, after digestion with a structure specific nuclease, accessible sites are mapped at sequence resolution along the entire RNA molecule which is used as a template for the reverse transcriptase elongation of a 5' end labelled selected primer (coding strand of a small restriction fragment of the cloned gene). This method circumvents any prior end-labelling of RNA, a technique with major limitations for large RNAs. In the second approach, a rapid "heterologous" sequencing can be easily applied to definite domains of an RNA molecule in a variety of species (or individuals), without additional DNA cloning nor end-labelling of RNA. By taking advantage of the presence of evolutionary conserved tracts within an RNA sequence, it allows a rapid analysis of RNA folding patterns in terms of phylogenetic comparisons : when located within such a conserved tract, selected restriction fragments from a cloned gene can be used as heterologous primers for sequencing the upstream divergent region in RNAs of other species by currently available technology, i.e. reverse transcriptase elongation in the presence of chain terminator dideoxynucleotides.

机译：我们已经开发出一种组合的方法来探测大RNA中的天然结构。在第一种方法中，用结构特异性核酸酶消化后，可访问的位点沿着整个RNA分子以序列分辨率定位，用作5'末端标记的选定引物（小链的编码链）逆转录酶延伸的模板克隆基因的限制性片段）。该方法规避了RNA的任何先前末端标记，这是对大型RNA的主要限制。在第二种方法中，快速的“异源”测序可轻松应用于各种物种（或个体）中RNA分子的确定域，而无需额外的DNA克隆或RNA的末端标记。通过利用RNA序列中进化保守区的存在，可以在系统发育比较方面快速分析RNA折叠模式：当位于此类保守区中时，可以从克隆基因中选择限制性片段用作异源引物，用于通过当前可用的技术对其他物种的RNA中的上游趋异区域进行测序，即在存在链终止剂双脱氧核苷酸的情况下逆转录酶延伸。

著录项

期刊名称 Nucleic Acids Research
作者
H L Qu; B Michot; J P Bachellerie;
展开▼
作者单位

展开▼
年(卷),期 1983(11),17
年度 1983
页码 5903–5920
总页数 18
原文格式 PDF
正文语种
中图分类分子生物学;
关键词

相似文献

外文文献

1. Conformation of yeast 18S rRNA. Direct chemical probing of the 5′ domain in ribosomal subunits and in deproteinized RNA by reverse transcriptase mapping of dimethyl sulfate-accessible sites [J] . Bernard Ehresmann, Chantal Ehresmann, Jean-Pierre Bachellerie, Nucleic acids research . 1985,第23期

机译：酵母18S rRNA的构象。通过硫酸二甲酯可及位点的逆转录酶作图法直接化学探测核糖体亚基和去蛋白RNA中的5'结构域
2. The 3′-terminal primary structure of five eukaryotic 18S rRNAs determined by the direct chemical method of sequencing. The highly conserved sequence include an invariant region complementary to eukaryotic 5S rRNA [J] . Ahmed A. Azad, Nicholas J. Deacon Nucleic acids research . 1980,第19期

机译：通过直接化学测序方法确定了五个真核18S rRNA的3'-末端一级结构。高度保守的序列包括与真核5S rRNA互补的恒定区
3. The 3′-terminal primary structure of five eukaryotic 18S rRNAs determined by the direct chemical method of sequencing. The highly conserved sequence include an invariant region complementary to eukaryotic 5S rRNA [J] . Ahmed A. Azad, Nicholas J. Deacon Nucleic acids research . 1980,第19期

机译：通过直接化学测序方法确定了五个真核18S rRNA的3'-末端一级结构。高度保守的序列包括与真核5S rRNA互补的恒定区
4. The 3-terminal primary structure of five eukaryotic 18S rRNAs determined by the direct chemical method of sequencing. The highly conserved sequences include an invariant region complementary to eukaryotic 5S rRNA. [O] . A A Azad, N J Deacon 1980

机译：通过直接化学测序方法确定的5个真核18S rRNA的3末端一级结构。高度保守的序列包括与真核5S rRNA互补的恒定区。
5. The 3'-terminal region of bacterial 23S ribosomal RNA: structure and homology with the 3'-terminal region of eukaryotic 28S rRNA and with chloroplast 4.5s rRNA. [O] . Machatt, M A, Ebel, J P, Branlant, C 1981

机译：细菌23S核糖体RNA的3'末端区域：与真核28S rRNA的3'末端区域和叶绿体4.5s rRNA的结构和同源性。

Improved methods for structure probing in large RNAs: a rapid heterologous sequencing approach is coupled to the direct mapping of nuclease accessible sites. Application to the 5 terminal domain of eukaryotic 28S rRNA.

摘要

著录项

相似文献

相关主题

期刊订阅