Nagao Y, Yagi M, Tsuchiya T, Tsuda M
Faculty of Pharmaceutical Sciences, Okayama University, Japan.
Nucleic Acids Symp Ser. 1988(19):89-92.
A quantitative procedure involving RNA-RNA hybridization kinetics was developed for measurement of specific mRNA accumulated in particular tissues and cells. Two types of riboprobes for quantitating mouse beta-tubulin mRNA were prepared; one was a truncated RNA covering only the coding portion of beta-tubulin cDNA and the other was a non-truncated RNA covering the vector portion as well as the coding portion. These antisense RNAs were hybridized with the mouse brain RNA, yielding heat-stable hybrids. The truncated and non-truncated antisense RNA probes showed similar hybridization kinetics. Hybridization of the sense RNA, consisting of the beta-tubulin coding portion, with the antisense RNA probe gave standards for determining the proportion of beta-tubulin mRNA in total brain RNA. By this method, the amounts of beta-tubulin mRNA included in the brains of mice of 10 and 50 days old were quantitated.
开发了一种涉及RNA-RNA杂交动力学的定量方法，用于测量特定组织和细胞中积累的特定mRNA。制备了两种用于定量小鼠β-微管蛋白mRNA的核糖探针；一种是仅覆盖β-微管蛋白cDNA编码部分的截短RNA，另一种是覆盖载体部分以及编码部分的非截短RNA。这些反义RNA与小鼠脑RNA杂交，产生热稳定的杂交体。截短和非截短的反义RNA探针显示出相似的杂交动力学。由β-微管蛋白编码部分组成的正义RNA与反义RNA探针杂交，为确定总脑RNA中β-微管蛋白mRNA的比例提供了标准。通过这种方法，对10日龄和50日龄小鼠脑中所含的β-微管蛋白mRNA量进行了定量。
