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经生长素处理大豆下胚轴后浓度降低的聚腺苷酸化RNA序列。

Polyadenylated RNA sequences which are reduced in concentration following auxin treatment of soybean hypocotyls.

作者信息

Baulcombe D C, Key J L

出版信息

J Biol Chem. 1980 Sep 25;255(18):88907-13.

PMID:6157689
Abstract

Previous work has shown that any effect of exogenous auxin on gene expression in soybean hypocotyl tissue must be restricted to a relatively small fraction of the polyadenylated RNA. However, kinetic hybridization analysis with cDNA probes revealed that a minor abundant class of sequences is markedly reduced in concentrations in the auxin-treated polyadenylated RNA. Recombinant plasmids containing copies of polyadenylated RNA species were constructed using the G-C tailing procedure and clones of auxin-regulated sequences were detected by differential in situ hybridization with cDNA of polyadenylated RNA from auxin-treated or untreated hypocotyls. Although the 12 clones which were selected all contained different size inserts, and were therefore independent, 11 of these apparently hybridized to just two different RNA species. The rate constant of the auxin-sensitive abundant component of the untreated polyadenylated RNA/DNA hybridization was similar to that of the reaction between the two major groups of clones and untreated polyadenylated RNA. This indicates that these cloned sequences are homologous with that cDNA fraction. The twelfth clone is thought to be representative of a group of less abundnt auxin-regulated polyadenylated mRNA species which had been detected in an earlier analysis of the in vitro translation products of soybean hypocotyl RNA. Both the timing and the extent of the influence of auxin on the relative concentration of these cloned sequences are quite consistent with a close relationship between growth regulation by auxin and its effects on gene expression.

摘要

先前的研究表明,外源生长素对大豆下胚轴组织基因表达的任何影响都必须局限于多聚腺苷酸化RNA的相对较小部分。然而,用cDNA探针进行的动力学杂交分析表明,一类少量丰富的序列在生长素处理的多聚腺苷酸化RNA中的浓度显著降低。使用G-C加尾程序构建了含有多聚腺苷酸化RNA物种拷贝的重组质粒,并通过与生长素处理或未处理的下胚轴多聚腺苷酸化RNA的cDNA进行差异原位杂交来检测生长素调节序列的克隆。虽然所选的12个克隆都包含不同大小的插入片段,因此是独立的,但其中11个显然只与两种不同的RNA物种杂交。未处理的多聚腺苷酸化RNA/DNA杂交中生长素敏感的丰富成分的速率常数与两组主要克隆与未处理的多聚腺苷酸化RNA之间反应的速率常数相似。这表明这些克隆序列与该cDNA片段同源。第十二个克隆被认为代表了一组丰度较低的生长素调节的多聚腺苷酸化mRNA物种,这些物种在早期对大豆下胚轴RNA体外翻译产物的分析中已被检测到。生长素对这些克隆序列相对浓度的影响的时间和程度与生长素的生长调节及其对基因表达的影响之间的密切关系相当一致。

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