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噬菌体及相关颗粒的琼脂糖凝胶电泳

Agarose gel electrophoresis of bacteriophages and related particles.

作者信息

Serwer P

出版信息

J Chromatogr. 1987 Jul 17;418:345-57. doi: 10.1016/0378-4347(87)80014-2.

Abstract

Viruses and related particles have been fractionated by electrophoresis through gels. For agarose gels, the radius at the exclusion limit for spheres varies from 1500 nm in a 0.04% gel to 3.6 nm in a 4.0% gel. Thus, the size of the gel's pores can be adjusted to sieve all known viruses. By measurement of electrophoretic mobility (mu) as a function of agarose concentration, the mu in the absence of a solid support (mu 0) can be determined for any particle. From the shape of a semilogarithmic plot of mu as a function of agarose percentage, a rod-shaped particle can be discriminated from a spherical particle. The sphere's radius can be determined from this plot with an accuracy of +/- 8%. Accuracy of +/- 1% has been more recently achieved using two-dimensional agarose gel electrophoresis. Though bacteriophages have been the primary object of study, the above techniques of agarose gel electrophoresis have also been applied to plant viruses and should be applicable to animal viruses. The mu 0 values measured for bacteriophages with and without their tail fibers suggest a mechanism of controlling attachment to a host. A related mechanism is proposed for the control of the virulence of animal viruses. Measurement of outer radius for different forms of the capsid of bacteriophage P22 reveals variability in outer radius too small to be detected by electron microscopy.

摘要

病毒及相关颗粒已通过凝胶电泳进行分级分离。对于琼脂糖凝胶,球形颗粒的排阻极限半径在0.04%的凝胶中为1500纳米,在4.0%的凝胶中为3.6纳米。因此,可以调整凝胶的孔径来筛分所有已知病毒。通过测量电泳迁移率(μ)作为琼脂糖浓度的函数,可以确定任何颗粒在没有固体支持物时的迁移率(μ0)。根据μ作为琼脂糖百分比的半对数图的形状,可以区分棒状颗粒和球形颗粒。通过该图可以确定球形颗粒的半径,精度为±8%。最近,使用二维琼脂糖凝胶电泳已实现±1%的精度。尽管噬菌体一直是主要的研究对象,但上述琼脂糖凝胶电泳技术也已应用于植物病毒,并且应该适用于动物病毒。对有尾纤维和无尾纤维的噬菌体测量的μ0值表明了一种控制与宿主附着的机制。有人提出了一种控制动物病毒毒力的相关机制。对噬菌体P22不同形式衣壳的外半径测量显示,外半径的变化太小,无法通过电子显微镜检测到。

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