从禽肉瘤病毒QV2中纯化病毒蛋白。

Purification of viral proteins from avian sarcoma virus QV2.

作者信息

Ueno A, Kohno M, Ishihama A, Toyoshima K

出版信息

J Biochem. 1979 Oct;86(4):929-42. doi: 10.1093/oxfordjournals.jbchem.a132625.

DOI:10.1093/oxfordjournals.jbchem.a132625

Abstract

A procedure was established whereby most of the major viral proteins were isolated to apparent homogeneity in biologically and immunologically active forms from a single batch of avian sarcoma virus QV2. For the initial step of purification, gently disrupted virions were fractionated by CsCl centrifugation into envelope proteins, RNA-dependent DNA polymerase, and viral core proteins. Further purification of envelope glycoproteins and DNA polymerase was performed by affinity chromatography on agarose columns cross-linked with plant lectins and poly(C), respectively. On the other hand, core proteins were fractionated by a combination of gel filtration and ion-exchange column chromatography into components p27, p19, and p15. The core protein p15 thus isolated retained proteolytic activity even after storage for 6 months. The present study also demonstrated that QV2 p19 is structurally altered from the corresponding protein of avian myeloblastosis virus (AMV), a reference avian leukosis-sarcoma virus having a well-characterized polypeptide composition.

摘要

建立了一种方法，通过该方法可以从一批禽肉瘤病毒QV2中以生物学和免疫活性形式分离出大部分主要病毒蛋白，达到明显的均一性。在纯化的第一步中，将温和破碎的病毒粒子通过氯化铯离心分离为包膜蛋白、依赖RNA的DNA聚合酶和病毒核心蛋白。包膜糖蛋白和DNA聚合酶的进一步纯化分别通过与植物凝集素和聚（C）交联的琼脂糖柱进行亲和层析来完成。另一方面，核心蛋白通过凝胶过滤和离子交换柱层析相结合的方法分离成p27、p19和p15组分。如此分离得到的核心蛋白p15即使在储存6个月后仍保留蛋白水解活性。本研究还表明，QV2 p19在结构上与禽成髓细胞瘤病毒（AMV）的相应蛋白不同，AMV是一种具有明确多肽组成的参考禽白血病-肉瘤病毒。