Chia L L, McLaughlin C
Mol Gen Genet. 1979 Feb 26;170(2):137-44. doi: 10.1007/BF00337788.
The decay kinetics of mRNA was studied in a yeast temperature-sensitive mutant, ts136, which is defective in cytoplasmic RNA production at 37 degree C. The disappearance of the synthetic capacity of mRNA was determined by withdrawing equal volumes of ts136 cell culture and pulse-labelling with [35S]methionine at various time intervals after the shift to 37 degrees C from 23 degrees C. The synthesized proteins were separated on a two-dimensional gel electrophoretic system and then quantitatively analyzed for theri incorporated radioactivities by scintillation counting. Our results show that yeast mRNAs have divergent functional half-lives ranging from 4.5 to 41 min, with an average value of 22 min. Each mRNA exhibits a simple exponential decay with its own characteristic dacay pattern. Of the approximately 500 major polypeptides made by yeast cells, which are detectable on autoradiograms of the gels, 80 were arbitrarily selected and the mRNAs coding for those polypeptides were examined for their decay kinetics.
在一个酵母温度敏感突变体ts136中研究了mRNA的衰变动力学,该突变体在37℃时细胞质RNA产生存在缺陷。通过在从23℃转移到37℃后的不同时间间隔抽取等体积的ts136细胞培养物并用[35S]甲硫氨酸进行脉冲标记,来测定mRNA合成能力的消失情况。合成的蛋白质在二维凝胶电泳系统上进行分离,然后通过闪烁计数对其掺入的放射性进行定量分析。我们的结果表明,酵母mRNA具有不同的功能半衰期,范围从4.5分钟到41分钟,平均值为22分钟。每个mRNA都呈现出具有自身特征衰变模式的简单指数衰变。在酵母细胞产生的大约500种主要多肽中,这些多肽在凝胶的放射自显影片上是可检测到的,从中任意选择了80种,并检查了编码这些多肽的mRNA的衰变动力学。