Weir E M, Riezman H, Grienenberger J M, Becker W M, Leaver C J
Eur J Biochem. 1980 Dec;112(3):469-77.
The relative levels of translatable messenger RNA for isocitrate lyase and malate synthase were determined in the dry seed and for the first seven days of development of cucumber cotyledons. After extraction and quantification of total and poly(A)-rich RNA each day, the RNA fractions were translated in an optimized wheat germ system and the specific polypeptides were immunoprecipitated quantitatively. The radiolabeled isocitrate lyase and malate synthase polypeptides were then fractionated on dodecylsulphate/polyacrylamide gels, visualized by exposure to X-ray film and quantified densitometrically. The relative levels of translatable messenger RNA for these enzymes rise and fall with a developmental program similar to the enzyme activities, but preceding the latter by about one day. This implies that the rise in enzyme activity is dependent upon a prior postgerminative increase in translatable messenger RNA for the enzymes. These studies also suggest that messenger RNA levels may be regulated, at least in part, by light.
测定了黄瓜子叶干种子及发育前七天中异柠檬酸裂解酶和苹果酸合酶可翻译信使核糖核酸的相对水平。每天对总RNA和富含多聚腺苷酸的RNA进行提取和定量后，将RNA组分在优化的小麦胚系统中进行翻译，并对特定多肽进行定量免疫沉淀。然后将放射性标记的异柠檬酸裂解酶和苹果酸合酶多肽在十二烷基硫酸钠/聚丙烯酰胺凝胶上进行分离，通过曝光于X射线胶片进行可视化，并进行光密度定量。这些酶的可翻译信使核糖核酸的相对水平随着与酶活性相似的发育程序上升和下降，但比酶活性提前约一天。这意味着酶活性的上升依赖于发芽后这些酶的可翻译信使核糖核酸的预先增加。这些研究还表明，信使核糖核酸水平可能至少部分受光的调节。
