L'vov V M, Shvartsman A L, Gaĭtskhoki V S, Skobeleva N A, Frolova L Iu
Mol Biol (Mosk). 1981 Jul-Aug;15(4):845-56.
The distribution of the sequences coding for ceruloplasmin (CP) in rat liver heterogeneous nuclear RNA (hnRNA) was studied using highly specific CP cDNA as a hybridization probe. The content of CP-coding sequences in poly(A)-containing and poly(A)-free subfractions of hnRNA was shown to be respectively 1 and 27 equivalents of CP mRNA molecule per one hepatocyte. The gel electrophoresis of hnRNA under strongly denaturing conditions with the subsequent transfer of RNA to diazobenzyloxymethyl paper and hybridization with [32P]-cDNA probe showed that CP mRNA sequences were of multiple molecular weight distribution. In particular, 9.0, 6.6, 2.4 and 1.6 megadalton fractions of non-polyadenylate hnRNA carried CP-coding sequences while the only hand that hybridized to CP cDNA was detected in polyadenylated hnRNA. This band was of a molecular weight 1.1-1.2 megadaltons corresponding to that of cytoplasmic CP mRNA. The hybridization of high molecular weight hnRNA with full-length CP cDNA followed by the determination of the size of cDNA fragments protected against SI nuclease demonstrated that coding sequences of CP pre-mRNA are interrupted by intervening sequences.
使用高度特异性的铜蓝蛋白(CP)cDNA作为杂交探针,研究了大鼠肝脏不均一核RNA(hnRNA)中编码铜蓝蛋白(CP)的序列分布。结果表明,hnRNA中含poly(A)和不含poly(A)的亚组分中CP编码序列的含量分别为每个肝细胞1个和27个CP mRNA分子当量。在强变性条件下对hnRNA进行凝胶电泳,随后将RNA转移至重氮苄氧基甲基纸上,并与[32P] - cDNA探针杂交,结果显示CP mRNA序列具有多种分子量分布。特别是,非聚腺苷酸化hnRNA的9.0、6.6、2.4和1.6兆道尔顿级分携带CP编码序列,而在聚腺苷酸化hnRNA中仅检测到与CP cDNA杂交的条带。该条带的分子量为1.1 - 1.2兆道尔顿,与细胞质CP mRNA的分子量相对应。高分子量hnRNA与全长CP cDNA杂交,随后测定抗S1核酸酶保护的cDNA片段大小,结果表明CP前体mRNA的编码序列被间隔序列中断。