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利用克隆的cDNA对大豆一组热激蛋白的mRNA进行分析。

An analysis of mRNAs for a group of heat shock proteins of soybean using cloned cDNAs.

作者信息

Schöffl F, Key J L

出版信息

J Mol Appl Genet. 1982;1(4):301-14.

PMID:6896722
Abstract

Hybridization studies carried out with poly(A)+ RNA and its corresponding cDNA showed the presence of a new highly abundant RNA class after heat shock (hs) at 40 degrees C in soybean hypocotyl compared to tissue incubated under normal conditions at 28 degrees C. cDNA clones complementary to RNAs of this class were isolated; eleven clones were characterized and used in the analysis of these abundant RNAs. The most abundant hs-sequences were found to be 800-900 nucleotides in length and present in about 19,000 copies per cell. Extensive sequence homology among hs-induced RNAs was indicated by cross-hybridization of cDNA clones and by common protein patterns generated in hybrid release translations. The existence of at least two different nucleotide sequences common to several different hs poly(A)+ mRNAs was documented by different, nonoverlapping protein patterns obtained by in vitro synthesis with hybrid selected RNAs. Four clones contained a sequence common to mRNAs for at least 13 proteins of 15,000-18,000 daltons; another sequence common to mRNA for three to four proteins of 21,000-23,000 daltons was selected by one clone. Two other clones selected a major hs-protein of about 18,000 daltons. The mRNAs of these low molecular weight hs-proteins accumulated rapidly after induction at either 40 degrees C or 42.5 degrees C and decreased rapidly during subsequent incubation at 28 degrees C.

摘要

用聚腺苷酸加(poly(A)+)RNA及其相应的互补DNA(cDNA)进行的杂交研究表明,与在28℃正常条件下培养的组织相比,大豆下胚轴在40℃热激(hs)后出现了一类新的高丰度RNA。分离出了与这类RNA互补的cDNA克隆;对11个克隆进行了表征,并用于分析这些丰度高的RNA。发现最丰富的热激序列长度为800 - 900个核苷酸,每个细胞中约有19,000个拷贝。cDNA克隆的交叉杂交以及杂交释放翻译中产生的共同蛋白质模式表明热激诱导的RNA之间存在广泛的序列同源性。通过用杂交选择的RNA进行体外合成获得的不同的、不重叠的蛋白质模式证明了几种不同的热激聚腺苷酸加(poly(A)+)信使核糖核酸(mRNA)共有至少两种不同的核苷酸序列。四个克隆包含了至少13种分子量为15,000 - 18,000道尔顿的蛋白质的mRNA共有的序列;一个克隆选择了分子量为21,000 - 23,000道尔顿的三到四种蛋白质的mRNA共有的另一个序列。另外两个克隆选择了一种主要的分子量约为18,000道尔顿的热激蛋白。这些低分子量热激蛋白的mRNA在40℃或42.5℃诱导后迅速积累,并在随后28℃的培养过程中迅速减少。

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