MacDonald R J, Stary S J, Swift G H
J Biol Chem. 1982 Dec 25;257(24):14582-5.
We have cloned via recombinant DNA technology the mRNA sequence of rat pancreatic ribonuclease, and have determined the entire nucleotide sequence of the mature message. Clones bearing RNase sequences within a double-stranded complementary DNA library of rat pancreatic mRNA were initially detected by hybridization with size-fractionated rat pancreatic polyadenylated RNA that included mRNA 0.85 to 1.0 kilobase in length. Recombinant plasmids bearing RNase mRNA sequences were conclusively identified by comparison of the amino acid sequence of the encoded protein with the known amino acid sequence of rat RNase. RNase mRNA is 783 nucleotides in length, plus a poly(a) tail with an average length of 140 nucleotides, and contains long 5' and 3' noncoding regions relative to other pancreatic mRNAs. It encodes a secretory preRNase of 152 amino acid residues including a signal peptide of 25 amino acids.
我们通过重组DNA技术克隆了大鼠胰腺核糖核酸酶的mRNA序列,并确定了成熟信使的完整核苷酸序列。在大鼠胰腺mRNA的双链互补DNA文库中,携带核糖核酸酶序列的克隆最初是通过与大小分级的大鼠胰腺多聚腺苷酸化RNA杂交检测到的,该RNA包括长度为0.85至1.0千碱基的mRNA。通过将编码蛋白质的氨基酸序列与大鼠核糖核酸酶的已知氨基酸序列进行比较,最终确定了携带核糖核酸酶mRNA序列的重组质粒。核糖核酸酶mRNA长度为783个核苷酸,加上平均长度为140个核苷酸的聚腺苷酸尾巴,相对于其他胰腺mRNA,它含有较长的5'和3'非编码区。它编码一种由152个氨基酸残基组成的分泌型前核糖核酸酶,包括一个由25个氨基酸组成的信号肽。