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麻疹病毒感染产生的宿主DNA合成抑制因子(DSF)的纯化

Purification of host DNA synthesis-suppressing factor (DSF) produced by infection with measles virus.

作者信息

Yamamoto T K, Minagawa T, Iida H

出版信息

Jpn J Microbiol. 1976 Dec;20(6):499-505. doi: 10.1111/j.1348-0421.1976.tb01018.x.

Abstract

Host DNA synthesis-suppressing factor (DSF) produced into culture fluid of cloned HeLa cells (HeLa C-9) infected with a small plaque variant of Toyoshima strain of measles virus was purified by precipitation with ammonium sulfate, chromatography on CM-cellulose and DEAE-cellulose, and gel-filtration on Sephadex G-100 and G-200. The specific activity of the finally purified DSF was 302 units/mg of protein representing approximately 300-fold purification. The molecular weight of DSF was estimated to be about 55 000. By isoelectric focusing, two kinds of DSF having isoelectric points of 4.24 and 5.24 were detectable. The purified DSF was able to suppress host DNA synthesis of HeLa cells, continuous human lymphoid cells (NC-37), mouse L cells and Meth-A cells derived from an ascitic tumor of the mouse. The activity of the purified DSF was inactivated by heating at 56 C for 30 min or by treatment with trypsin.

摘要

从感染了麻疹病毒丰岛株小蚀斑变种的克隆海拉细胞(海拉C-9)培养液中产生的宿主DNA合成抑制因子(DSF),通过硫酸铵沉淀、CM-纤维素和DEAE-纤维素柱层析以及Sephadex G-100和G-200凝胶过滤进行纯化。最终纯化的DSF比活性为302单位/毫克蛋白质,纯化倍数约为300倍。DSF的分子量估计约为55000。通过等电聚焦可检测到两种等电点分别为4.24和5.24的DSF。纯化的DSF能够抑制海拉细胞、人连续淋巴细胞(NC-37)、小鼠L细胞以及源自小鼠腹水瘤的Meth-A细胞的宿主DNA合成。纯化的DSF活性在56℃加热30分钟或用胰蛋白酶处理后会失活。

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