Masamune Y, Richardson C C
J Biol Chem. 1977 Dec 10;252(23):8498-053.
Commercial preparations of guanosine 5'-triphosphate and deoxyguanosine 5'-triphosphate are contaminated with oligoribonucleotides 4 to 6 residues in length. The oligoribonucleotides can be separated from the nucleoside 5'-triphosphates by chromatography on DEAE-Sephadex A-25 or by gel filtration through Sephadex G-25. The oligoribonucleotides are effective primers for the DNA polymerase of bacteriophage T7 on the single-stranded circular DNAs of phage fd and phiX174; they are covalently attached to the 5' terminus of the newly synthesized DNA. The priming activity is specific; the oligoribonucleotides do not serve as primers for DNA polymerase I of Escherichia coli or for the DNA polymerase induced by phage T4.
鸟苷 5'-三磷酸和脱氧鸟苷 5'-三磷酸的商业制剂被长度为 4 至 6 个残基的寡核糖核苷酸污染。寡核糖核苷酸可通过在 DEAE-葡聚糖 A-25 上进行色谱分离或通过葡聚糖 G-25 凝胶过滤从核苷 5'-三磷酸中分离出来。这些寡核糖核苷酸是噬菌体 T7 的 DNA 聚合酶在噬菌体 fd 和 phiX174 的单链环状 DNA 上的有效引物;它们共价连接到新合成 DNA 的 5' 末端。引发活性具有特异性;这些寡核糖核苷酸不能作为大肠杆菌 DNA 聚合酶 I 或噬菌体 T4 诱导的 DNA 聚合酶的引物。