通过四磷酸盐测定法测量艾氏腹水癌细胞中初级转录本的大小。
Size of primary transcripts in Ehrlich ascites cells as measured by tetraphosphate determination.
作者信息
Schmincke C D, Herrmann K, Hausen P
出版信息
Proc Natl Acad Sci U S A. 1976 Jun;73(6):1994-8. doi: 10.1073/pnas.73.6.1994.
A method for the quantitation of 5"-tetraphosphate ends in 32P-labeled RNA has been developed. The tetraphosphate content of different RNA fractions obtained from Ehrlich ascites cells labeled with 32P for different lengths of time has been determined. Ribosomal RNA and poly(U)-binding RNA, labeled for long periods, (mRNA) lack 5'-terminal tetraphosphate. 5S RNA, pulse labeled 4-5S RNA, and poly(U)-binding hnRNA (heterogeneous nuclear RNA) do contain tetraphosphate. From the amount of the tetraphosphate, molecular weight data can be calculated for these RNA fractions which agree with independent determinations by denaturing gel electrophoresis. The results demonstrate that the majority of the poly(A) containing hnRNA molecules are small (less than 28S) and contain the tetraphosphate of the primary transcript. Therefore, they do not originate from the 3'-end of large molecules by processing events.
已开发出一种定量32P标记RNA中5'-四磷酸末端的方法。测定了从用32P标记不同时长的艾氏腹水细胞中获得的不同RNA组分的四磷酸含量。长时间标记的核糖体RNA和聚（U）结合RNA（mRNA）缺乏5'-末端四磷酸。5S RNA、脉冲标记的4-5S RNA和聚（U）结合的hnRNA（不均一核RNA）确实含有四磷酸。根据四磷酸的量，可以计算出这些RNA组分的分子量数据，这些数据与通过变性凝胶电泳的独立测定结果一致。结果表明，大多数含聚（A）的hnRNA分子较小（小于28S），并含有初级转录本的四磷酸。因此，它们不是通过加工事件从大分子的3'-末端产生的。
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