M13噬菌体在病毒感染的大肠杆菌提取物中的膜相关组装。
Membrane-associated assembly of M13 phage in extracts of virus-infected Escherichia coli.
作者信息
Wickner W, Killick T
出版信息
Proc Natl Acad Sci U S A. 1977 Feb;74(2):505-9. doi: 10.1073/pnas.74.2.505.
Abstract
Assembly of coliphage M13 is known to occur as the viral DNA crosses the cytoplasmic membrane, shedding its virus-coded DNA unwinding protein and acquiring from the membrane approximately 2400 copies of the major coat protein. Conditions are described in which extracts of M13-infected E. coli and membranes prepared from such extracts will support virus assembly at a rate equivalent to that of intact cells. Extracts prepared from cells infected with temperature-sensitive M13 mutants in genes 1, 3, 4, or 5 are temperature-sensitive in this cell-free assembly reaction. Phage assembly in vitro requires magnesium and as yet an unidentified heat-stable cofactor of low molecular weight. The rate of virus assembly is approximately linear with respect to extract concentration over a 10(4)-fold range, consistent with the observation that the entire M13 assembly activity copurifies with the cell membrane fraction.
摘要
已知噬菌体M13的组装发生在病毒DNA穿过细胞质膜时，此时它会脱落病毒编码的DNA解旋蛋白，并从膜上获取大约2400份主要衣壳蛋白。文中描述了这样的条件：用M13感染的大肠杆菌提取物以及由此制备的膜，能以与完整细胞相当的速率支持病毒组装。用基因1、3、4或5中温度敏感型M13突变体感染细胞制备的提取物，在这种无细胞组装反应中对温度敏感。体外噬菌体组装需要镁以及一种尚未鉴定出的低分子量热稳定辅因子。在10^4倍的范围内，病毒组装速率与提取物浓度大致呈线性关系，这与整个M13组装活性与细胞膜部分共纯化的观察结果一致。
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