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通过与伴刀豆球蛋白A(鼠白血病病毒-植物血凝素-弗氏病毒)凝集来纯化肿瘤病毒

Purification of oncornaviruses by agglutination with concanacalin A (murine leukemia virus-phytohemagglutinin-friend virus).

作者信息

Stewart M L, Summers D F, Soeiro R, Fields B N, Maizel J V

出版信息

Proc Natl Acad Sci U S A. 1973 May;70(5):1308-12. doi: 10.1073/pnas.70.5.1308.

Abstract

Concanavalin A (Con A) has been used to rapidly and selectively agglutinate murine and avian oncornavirions from culture medium or plasma. The agglutinated virus was concentrated rapidly and gently by low-speed centrifugation and solubilization with alpha-methyl mannoside. Infectious virus was purified 2.3 times with respect to nucleic-acid content, and more than 60% of its infectivity was recovered. Infectious particles of densities 1.18 and 1.16 g/cm(3) were found in mouse cells infected with Friend virus. Con A reacted only with particles of density 1.16 g/cm(3), indicating heterogeneity with respect to carbohydrate content or structure as well as buoyant density. Electron microscopy of virus agglutinated with Con A showed a zone of Con A-glycoprotein complexes averaging 12-15 nm in thickness.

摘要

伴刀豆球蛋白A(Con A)已被用于快速、选择性地凝集来自培养基或血浆中的鼠源和禽源肿瘤病毒粒子。通过低速离心和用α-甲基甘露糖苷溶解,可快速且温和地浓缩凝集的病毒。相对于核酸含量,感染性病毒被纯化了2.3倍,并且回收了超过60%的感染性。在感染了弗氏病毒的小鼠细胞中发现了密度为1.18和1.16 g/cm³的感染性颗粒。Con A仅与密度为1.16 g/cm³的颗粒发生反应,这表明在碳水化合物含量或结构以及浮力密度方面存在异质性。用Con A凝集的病毒的电子显微镜观察显示,Con A - 糖蛋白复合物区域的平均厚度为12 - 15纳米。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f3c4/433485/c16b96c873a3/pnas00068-0026-a.jpg

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