DEVELOPMENTAL REGULATION OF GENE-EXPRESSION IN WHEAT EMBRYOS - MOLECULAR-CLONING OF A DNA-SEQUENCE ENCODING THE EARLY METHIONINE-LABELED (EM) POLYPEPTIDE
DEVELOPMENTAL REGULATION OF GENE-EXPRESSION IN WHEAT EMBRYOS - MOLECULAR-CLONING OF A DNA-SEQUENCE ENCODING THE EARLY METHIONINE-LABELED (EM) POLYPEPTIDE
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DOI：
10.1111/j.1432-1033.1984.tb08561.x
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发表时间：
1984-01-01
期刊：
影响因子：
--
通讯作者：
CUMING, AC
中科院分区：
文献类型：
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作者：
CUMING, AC
早期甲硫氨酸标记的（Em）多肽是在胞质部分中发现的最丰富的单一多肽（30，000 × 106）。g上清液）。通过用[35 S]甲硫氨酸标记，在小麦胚萌发的最早阶段（0-3 h）容易检测到多肽的合成。此后，Em多肽的合成迅速下降。通过分子克隆与来自干小麦胚的mRNA互补的DNA，分离编码该多肽的一部分的DNA序列。该克隆序列的身份被证实的杂交选择翻译的EM信使，该产品被分析的1-和2-维聚丙烯酰胺凝胶电泳，并通过分析其部分蛋白水解消化产物。凝胶印迹杂交从胚胎中分离的RNA在连续阶段的萌发与此克隆的DNA序列证实，在Em多肽的合成，在体内标记检测到的下降，是其相应的mRNA的降解的结果。
The early-methionine-labeled (Em) polypeptide is the most abundant single polypeptide found in the cytosolic fraction (30,000 .times. g supernatant) of dry wheat embryos. Synthesis of the polypeptide is readily detectable during the earliest stages (0-3 h) of wheat embryo germination, by labeling with [35S]methionine. Thereafter, synthesis of the Em polypeptide declines rapidly. A DNA sequence encoding a portion of this polypeptide was isolated by the molecular cloning of DNA complementary to the mRNA from dry wheat embryos. The identity of this cloned sequence was confirmed by hybrid-selected translation of the Em messenger, the product being analyzed by 1- and 2-dimensional polyacrylamide gel electrophoresis, and by analysis of its partial proteolytic digestion products. Gel blot hybridization of RNA isolated from embryos at successive stages of germination with this cloned DNA sequence confirms that the decline in the synthesis of the Em polypeptide, detected by labeling in vivo, is a consequence of the degradation of its corresponding mRNA.