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鸡胚细胞单层培养中黏液病毒神经氨酸酶的细胞内合成。I. 新城疫病毒感染细胞中的神经氨酸酶活性、血凝素合成及细胞结合唾液酸含量

Intracellular synthesis of myxovirus neuraminidase in chick embryo cell monolayer culture. I. Neuraminidase activity, hemagglutinin synthesis, and content of cellbound sialic acid in Newcastle disease virus-infected cells.

作者信息

Lipkind M A, Tsvetkova I V

出版信息

J Virol. 1967 Apr;1(2):327-33. doi: 10.1128/JVI.1.2.327-333.1967.

Abstract

Neuraminidase (Nase) activity of chick embryo monolayer cell homogenates was determined by its rate of splitting of neuraminlactose, free neuraminic acid (NA) being determined by the thiobarbituric acid assay. Noninfected cells were found to have no detectable amount of Nase activity. Newcastle disease virus (NDV)-infected cells (multiplicity of infection, 20 to 75 plaque-forming units per cell) displayed a high level of Nase synthesis, the rate of synthesis being parallel to that of hemagglutinin (HA) synthesis (with a 1.5 hr delay in the latter). An "eclipse" of the Nase and HA activities associated with the virus that was adsorbed onto cells was observed. The data provide evidence that the Nase is not incorporated into the viral envelope from a pre-existing cell supply but that its synthesis is coded by the viral genome. The content of cell-bound sialic acid, determined simultaneously in infected-cell homogenates, showed characteristic features allowing certain conclusions concerning the renewal of NA-terminating cell receptors during the course of infection, and the intracellular action of the Nase of the virus introduced into cells by the inoculum and that of the newly synthesized Nase at different stages of infection.

摘要

通过神经氨酸乳糖的裂解速率来测定鸡胚单层细胞匀浆中的神经氨酸酶(Nase)活性,游离神经氨酸(NA)则通过硫代巴比妥酸测定法来测定。发现未感染的细胞没有可检测到的Nase活性。感染新城疫病毒(NDV)的细胞(感染复数为每细胞20至75个蚀斑形成单位)表现出高水平的Nase合成,合成速率与血凝素(HA)合成速率平行(后者延迟1.5小时)。观察到与吸附在细胞上的病毒相关的Nase和HA活性出现“隐蔽期”。这些数据证明,Nase并非从预先存在的细胞储备中整合到病毒包膜中,而是其合成由病毒基因组编码。在感染细胞匀浆中同时测定的细胞结合唾液酸含量显示出特征性,这使得我们能够就感染过程中NA末端细胞受体的更新以及接种物引入细胞的病毒Nase和新合成的Nase在感染不同阶段的细胞内作用得出某些结论。

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