Sippel A E, Groner B, Hynes N, Schütz G
Eur J Biochem. 1977 Jul 1;77(1):153-64. doi: 10.1111/j.1432-1033.1977.tb11653.x.
Total rat liver poly(A)-containing polysomal mRNA was size-fractionated on polyacrylamide gels in 98% formamide. Complementary DNA (cDNA) was prepared from the 8--14-S mRNA fraction and separated into sequences representing abundant and non-abundant mRNAs. The cDNA complementary to the abundant small mRNA of the rat liver cell (approximately 20 species) was hybridized to nuclear RNA of different lengths to determine the size distribution of nuclear RNA molecules which contain these messenger sequences. It was found that: 1. All abundant 8--14-S poly(A)-containing mRNAs have larger nuclear precursor molecules; 20% of the different messenger sequences are found in nuclear RNA of several times their cytoplasmic length. 2. 70% of the mass of the examined nuclear messenger sequences is in RNA molecules of a size similar to their polysomal mRNA; 30% are in larger than 18-S RNA and 2% are between 37 S and 44 S. 3. The majority of small messenger-containing RNA molecules in the RNA prepared from isolated nuclei are of true nuclear origin, since their frequency distribution differs significantly from that of the polysomal 8--14-S mRNA.
将大鼠肝脏中含多聚腺苷酸的多核糖体mRNA在98%甲酰胺的聚丙烯酰胺凝胶上进行大小分级分离。从8 - 14S的mRNA组分制备互补DNA(cDNA),并将其分离为代表丰富和不丰富mRNA的序列。将与大鼠肝细胞丰富的小mRNA(约20种)互补的cDNA与不同长度的核RNA杂交,以确定包含这些信使序列的核RNA分子的大小分布。结果发现:1. 所有丰富的含多聚腺苷酸的8 - 14S mRNA都有更大的核前体分子;20%的不同信使序列存在于长度为其细胞质长度数倍的核RNA中。2. 所检测的核信使序列质量的70%存在于大小与其多核糖体mRNA相似的RNA分子中;30%存在于大于18S的RNA中,2%存在于37S和44S之间。3. 从分离的细胞核制备的RNA中,大多数含小信使的RNA分子是真正的核源,因为它们的频率分布与多核糖体8 - 14S mRNA的频率分布有显著差异。