Raj N B, Pitha P M
Proc Natl Acad Sci U S A. 1981 Dec;78(12):7426-30. doi: 10.1073/pnas.78.12.7426.
The levels of interferon mRNA as a function of interferon induction by poly(rI) . poly(rC) in human fibroblast cells were determined by RNA hybridization using a cloned beta interferon cDNA and by translation in Xenopus oocytes. Whereas previous studies analyzed mixtures of interferons, the availability of the cloned beta interferon cDNA and the antiserum to purified beta interferon enabled us to focus on the expression of only one class (beta) of interferon genes. The induction of interferon synthesis depends primarily on the accumulation of interferon beta mRNA in the cells, and the interferon beta mRNA rapidly disappears several hours after its appearance in the cytoplasm. No detectable interferon beta mRNA sequences are present in uninduced cells. The degradation of interferon beta mRNA in the induced cells requires ongoing protein synthesis; accumulation of interferon beta mRNA was observed in the continuous presence of cycloheximide. The interferon beta mRNA detected at the early stages of induction is 1100 nucleotides long and its size progressively decreases with time. By both the hybridization and the translational assay in Xenopus oocytes, only one size of interferon beta mRNA and one species of beta interferon could be identified.
利用克隆的β干扰素cDNA通过RNA杂交以及在非洲爪蟾卵母细胞中进行翻译,测定了人成纤维细胞中作为聚肌苷酸-聚胞苷酸诱导干扰素作用函数的干扰素mRNA水平。以往的研究分析的是干扰素混合物,而克隆的β干扰素cDNA和针对纯化β干扰素的抗血清使我们能够专注于仅一类(β)干扰素基因的表达。干扰素合成的诱导主要取决于细胞中干扰素βmRNA的积累,并且干扰素βmRNA在出现在细胞质中数小时后迅速消失。未诱导的细胞中不存在可检测到的干扰素βmRNA序列。诱导细胞中干扰素βmRNA的降解需要持续的蛋白质合成;在放线菌酮持续存在的情况下观察到了干扰素βmRNA的积累。在诱导早期检测到的干扰素βmRNA长度为1100个核苷酸,其大小随时间逐渐减小。通过在非洲爪蟾卵母细胞中的杂交和翻译测定,只能鉴定出一种大小的干扰素βmRNA和一种β干扰素。