Plesner P, Goodchild J, Kalckar H M, Zamecnik P C
Proc Natl Acad Sci U S A. 1987 Apr;84(7):1936-9. doi: 10.1073/pnas.84.7.1936.
Endogenous oligonucleotides were found in trichloroacetic acid extracts of hamster lung fibroblasts and Tetrahymena cells. Peaks of radioactivity that eluted with retention times similar to oligonucleotide markers (5- to 50-mer) were found by HPLC in cells labeled briefly with 32Pi. Only minute amounts of UV-absorbing material were detected, consistent with a rapid turnover of phosphate groups. The 32P-labeled material also migrated as oligonucleotides on 20% polyacrylamide gels; it was not hydrolyzed by alkaline phosphatase but was digested by snake venom phosphodiesterase, S1 nuclease, and pancreatic RNase and was phosphorylated by T4 polynucleotide kinase. The 32P-labeled material isolated by HPLC was alkali labile and the hydrolyzate ran as nucleotides on paper chromatography. It is concluded that the oligonucleotides are mainly oligoribonucleotides, but it is possible that oligodeoxynucleotides are also present.
在仓鼠肺成纤维细胞和四膜虫细胞的三氯乙酸提取物中发现了内源性寡核苷酸。在用32Pi短暂标记的细胞中,通过高效液相色谱法(HPLC)发现,放射性峰的洗脱保留时间与寡核苷酸标记物(5至50聚体)相似。仅检测到微量的紫外线吸收物质,这与磷酸基团的快速周转一致。32P标记的物质在20%聚丙烯酰胺凝胶上也以寡核苷酸的形式迁移;它不被碱性磷酸酶水解,但被蛇毒磷酸二酯酶、S1核酸酶和胰核糖核酸酶消化,并被T4多核苷酸激酶磷酸化。通过HPLC分离的32P标记物质对碱不稳定,其水解产物在纸色谱上呈核苷酸形式迁移。结论是这些寡核苷酸主要是寡核糖核苷酸,但也可能存在寡脱氧核苷酸。