Ryffel G U, McCarthy B J
Biochemistry. 1975 Apr 8;14(7):1379-85. doi: 10.1021/bi00678a006.
The kinetics of hybridization of polyadenylated RNA from mouse L-cells with complementary DNA (cDNA) synthesized with reverse transcriptase revealed three classes of differing abundance. The simplest interpretation requires three frequency classes representing polyadenylated RNA; 5, 45, and 50 percent of the total polyadenylated RNA and about 3, 300, and 7600 different RNA sequences of 6 times 10-5 daltons, respectively. The complementary DNA synthesized with L-cell polyadenylated RNA as template hybridized efficiently with RNA from different mouse tissues, indicating that most species of the L-cell RNA in the highand middle frequency class are present in all mouse tissues. Kinetics of hybridization of complementary DNA synthesized with cytoplasmic polyadenylated brain RNA as template suggested a higher complexity for brain RNA. Thirty-five percent of this brain cDNA failed to hybridize with L-cell RNA. This complementary DNA fraction, isolated by hydroxylapatite chromatography, represented approximately 11,000 RNA sequences specific for the brain. On the other hand, hybridization of complementary DNA synthesized on polyadenylated mouse liver RNA with L-cell RNA failed to demonstrate differences between these two groups of polyadenylated RNA.
从小鼠L细胞中提取的多聚腺苷酸化RNA与用逆转录酶合成的互补DNA(cDNA)杂交的动力学显示出三类丰度不同的情况。最简单的解释是,这三类频率代表了多聚腺苷酸化RNA;分别占总多聚腺苷酸化RNA的5%、45%和50%,以及约3种、300种和7600种分子量为6×10⁻⁵道尔顿的不同RNA序列。以L细胞多聚腺苷酸化RNA为模板合成的互补DNA能有效地与来自不同小鼠组织的RNA杂交,这表明高频和中频类别的大多数L细胞RNA物种存在于所有小鼠组织中。以细胞质多聚腺苷酸化脑RNA为模板合成的互补DNA的杂交动力学表明脑RNA的复杂性更高。该脑cDNA的35%不能与L细胞RNA杂交。通过羟基磷灰石层析分离得到的这部分互补DNA代表了大约11000种脑特异性RNA序列。另一方面,以多聚腺苷酸化小鼠肝RNA合成的互补DNA与L细胞RNA杂交,未显示出这两组多聚腺苷酸化RNA之间的差异。