Sehgal P B, Soreq H, Tamm I
Proc Natl Acad Sci U S A. 1978 Oct;75(10):5030-3. doi: 10.1073/pnas.75.10.5030.
Polynucleotide phosphorylase (polyribonucleotide:orthophosphate nucleotidyltransferase, EC 2.7.7.8) purified from Escherichia coli was used enzymatically to deadenylate polyadenylated human fibroblast interferon mRNA preparations obtained from human diploid fibroblasts (FS-4 strain) induced by poly(I)-poly(C) (20 microgram/ml) in the presence of cycloheximide (50 microgram/ml, 4 hr). Both the polyadenylated and the deadenylated interferon mRNA preparations were translated into biologically active human interferon when injected into oocytes of Xenopus laevis. In the oocytes the functional stability of deadenylated interferon mRNA was indistinguishable from that of polyadenylated interferon mRNA.
从大肠杆菌中纯化得到的多核苷酸磷酸化酶(聚核糖核苷酸:正磷酸核苷酸基转移酶,EC 2.7.7.8)被用于酶促反应,使从人二倍体成纤维细胞(FS-4株)中获得的聚腺苷酸化人成纤维细胞干扰素mRNA制剂去腺苷酸化。这些人二倍体成纤维细胞由聚(I)-聚(C)(20微克/毫升)在环己酰亚胺(50微克/毫升,4小时)存在的情况下诱导产生。当将聚腺苷酸化和去腺苷酸化的干扰素mRNA制剂注射到非洲爪蟾的卵母细胞中时,二者均可被翻译成具有生物活性的人干扰素。在卵母细胞中,去腺苷酸化的干扰素mRNA的功能稳定性与聚腺苷酸化的干扰素mRNA的功能稳定性没有差异。