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玉米胚乳蔗糖合酶mRNA的体外翻译

Translation of Zea mays endosperm sucrose-synthase mRNA in vitro.

作者信息

Wöstemeyer J, Behrens U, Merckelbach A, Müller M, Starlinger P

出版信息

Eur J Biochem. 1981;114(1):39-44. doi: 10.1111/j.1432-1033.1981.tb06168.x.

Abstract

mRNA of 23-day-old maize endosperm was translated both in wheat germ extracts and rabbit reticulocyte lysates. A protein with an apparent molecular weight of 88,000 comigrates in dodecylsulfate/polyacrylamide electrophoresis with sucrose synthase. This protein is precipitated with an antiserum against sucrose synthase and shows the same protease digestion pattern as the enzyme. It is not synthesized with mRNA extracted from sh/sh mutant kernels lacking sucrose synthase. By these criteria, the protein is the translation product in vitro of sucrose synthase mRNA. The separation of mRNA in methylmercury-hydroxide--agarose gels and subsequent translation indicates a length of sucrose synthase mRNA of 2800 nucleotides which is compatible with the coding length necessary for a protein with a molecular weight of 88,000 plus untranslated sequences.

摘要

23日龄玉米胚乳的mRNA在小麦胚芽提取物和兔网织红细胞裂解物中均能被翻译。一种表观分子量为88,000的蛋白质在十二烷基硫酸盐/聚丙烯酰胺电泳中与蔗糖合酶迁移率相同。该蛋白质能被抗蔗糖合酶的抗血清沉淀,并且显示出与该酶相同的蛋白酶消化模式。它不能用从缺乏蔗糖合酶的sh/sh突变体籽粒中提取的mRNA合成。根据这些标准,该蛋白质是蔗糖合酶mRNA的体外翻译产物。在氢氧化甲基汞-琼脂糖凝胶中分离mRNA并随后进行翻译表明,蔗糖合酶mRNA的长度为2800个核苷酸,这与一个分子量为88,000的蛋白质加上非翻译序列所需的编码长度相符。

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