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通过荧光猝灭测量流感病毒在细胞内酸性区室中的融合。

Fusion of influenza virus in an intracellular acidic compartment measured by fluorescence dequenching.

作者信息

Stegmann T, Morselt H W, Scholma J, Wilschut J

机构信息

Laboratory of Physiological Chemistry, University of Groningen, The Netherlands.

出版信息

Biochim Biophys Acta. 1987 Nov 2;904(1):165-70. doi: 10.1016/0005-2736(87)90100-3.

Abstract

The fusion of influenza virus with cultured cells has been investigated. The virus was labelled with the fluorescent probe octadecyl rhodamine B and fusion was monitored as fluorescence dequenching due to dilution of the probe from the viral into a cellular target membrane. Fusion with the plasma membrane does not occur, unless the extracellular pH is temporarily lowered. At neutral pH fusion occurs only after a lag phase of 10-15 min, the time required for virus internalization, and the reaction is inhibited by NH4Cl, indicating that it takes place in an intracellular acidic compartment, most likely the endosome. This suggests that influenza virus infects cells via the endocytic pathway.

摘要

对流感病毒与培养细胞的融合进行了研究。该病毒用荧光探针十八烷基罗丹明B进行标记,由于探针从病毒稀释到细胞靶膜中导致荧光猝灭,以此监测融合情况。除非细胞外pH值暂时降低,否则不会与质膜发生融合。在中性pH值下,融合仅在10 - 15分钟的延迟期后发生,这是病毒内化所需的时间,并且该反应受到氯化铵的抑制,表明其发生在细胞内酸性区室中,很可能是内体。这表明流感病毒通过内吞途径感染细胞。

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