Timchenko N A, Timchenko L T, Bilobrov S V, Zhuchenko O P, Krutiakov V M
Mol Biol (Mosk). 1987 May-Jun;21(3):640-6.
A library of double-stranded cDNA was prepared using poly (A) + RNA from regenerating rat liver 20 h after partial hepatectomy. Differential screening of 350 recombinant clones with cDNA-G0 and cDNA-S identified eleven cDNA clones (pRL), the sequences of which were preferentially expressed during the DNA replication period. Levels of mRNAs complementary to these clones were 2--10-fold higher in the S-period, than in G0. Using plasmid cDNAs to different mRNA, pRL we have investigated the changes in the levels of mRNA pRL during liver regeneration. The level of mRNA mRL2 and pRL79 was increased just before DNA replication. mRNA pRL35 accumulates after partial hepatectomy with the maximum at 6 h. The augment of two other mRNA concentrations was expressed to a lesser extent. Northern-blot analysis allowed to determine the individual dual mRNAs corresponding to each of the three clones with their sizes ranging from about 1650 to 3900 bases. Three mRNAs (pRL35, 67 and 79) were shown (by hybrid-selected translation) to code for proteins of about 100, 140 and 120 kDa, respectively.
利用部分肝切除术后20小时再生大鼠肝脏的聚腺苷酸（poly (A) +）RNA制备双链cDNA文库。用cDNA-G0和cDNA-S对350个重组克隆进行差异筛选，鉴定出11个cDNA克隆（pRL），其序列在DNA复制期优先表达。与这些克隆互补的mRNA水平在S期比在G0期高2至10倍。利用针对不同mRNA的质粒cDNA（pRL），我们研究了肝脏再生过程中mRNA pRL水平的变化。mRNA mRL2和pRL79的水平在DNA复制前升高。mRNA pRL35在部分肝切除术后积累，在6小时达到最大值。另外两种mRNA浓度的增加程度较小。Northern印迹分析能够确定与三个克隆中的每一个相对应的单个双链mRNA，其大小约为1650至3900个碱基。通过杂交选择翻译显示，三种mRNA（pRL35、67和79）分别编码约100、140和120 kDa的蛋白质。
