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经同位素标记的双链f2噬菌体RNA诱导产生的小鼠细胞干扰素的纯化

Purification of mouse cell interferon induced by isotopically labelled double-stranded f2 phage RNA.

作者信息

Táborský I, Doskocil J, Stancek D

出版信息

J Hyg Epidemiol Microbiol Immunol. 1977;21(3):261-70.

PMID:564366
Abstract

The dynamics of interferon formation by an established cell line of mouse fibroblast (L cells) and by mouse peritoneal leukocytes induced by double-stranded RNA extracted from E. coli f2 phage is described. The L cells produced interferon at a lower rate, the maximum values were obtained at 12 to 20 hours after induction, and the production was ultimately dependent on the established cell line used and on the presence of DEAE-dextran during induction. The mouse peritoneal leukocytes (MPL), on the other hand, did not require DEAE-dextran and the maximum of interferon production was reached between 6 and 12 hours after induction. Both the L cell- and the MPL-interferons were purified and concentrated so that the final specific biologic activity was 100-to 300-fold higher than that of the initial preparations (1 to 5 X 10(6) interferon units per mg protein). Polyacrylamide gel electrophoresis showed similar migration profiles for the preparations of both interferons. The smaller part of the activity was situated in a broader, slow-moving peak and the greater part formed a sharp, high and fast-moving peak. Using 3H uridine-labelled f2 ds-RNA for induction of interferon it was found that one of the radioactivity zones coincided with the fast-moving activity peak of the purified and concentrated interferon.

摘要

本文描述了由已建立的小鼠成纤维细胞系(L细胞)以及由从大肠杆菌f2噬菌体中提取的双链RNA诱导的小鼠腹腔白细胞产生干扰素的动力学过程。L细胞产生干扰素的速率较低,诱导后12至20小时达到最大值,其产生最终取决于所用的已建立细胞系以及诱导过程中DEAE-葡聚糖的存在。另一方面,小鼠腹腔白细胞(MPL)不需要DEAE-葡聚糖,诱导后6至12小时达到干扰素产生的最大值。L细胞干扰素和MPL干扰素均经过纯化和浓缩,最终的比生物活性比初始制剂高100至300倍(每毫克蛋白质1至5×10⁶个干扰素单位)。聚丙烯酰胺凝胶电泳显示两种干扰素制剂的迁移图谱相似。活性的较小部分位于较宽的、移动缓慢的峰中,较大部分形成尖锐的、高的且移动快速的峰。使用³H尿苷标记的f2双链RNA诱导干扰素时,发现其中一个放射性区域与纯化和浓缩干扰素的快速移动活性峰重合。

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