Li J K
Department of Biology, Utah State University, Logan 84322-5500.
J Virol Methods. 1987 Sep;17(3-4):299-310. doi: 10.1016/0166-0934(87)90140-6.
The seven mouse mammary tumor virus (MMTV) structural polypeptides (GP52, GP36, P28, P23, P14, P12, and P10) were purified to apparent homogeneity. An improved and reproducible chromatographic method was used to obtain high yields of viral proteins from the same batch of isopycnically purified MMTV. The viral structural proteins were first extracted repeatedly with a high salt and high pH buffer in the presence of 1% Triton X-100. Loss of the minor viral proteins during column purification was minimized by first purifying the smaller molecular weight viral proteins (P14, P12 and P10) by oligo-d(T) column chromatography from the dissociated virions. The other four major viral polypeptides (GP52, GP36, P28 and P23) were then fractionated by affinity and ion-exchange columns. High titer polyclonal antibodies were produced against all of the seven structural proteins except P12 and P10. These antisera were monospecific and showed no cross-reactivity in radioimmunoassay towards other MMTV proteins. With minor modifications, this method has also been applied to purify other structural proteins from several different C-type retroviruses.
七种小鼠乳腺肿瘤病毒（MMTV）结构多肽（GP52、GP36、P28、P23、P14、P12和P10）被纯化至表观均一性。采用一种改进的、可重复的色谱方法，从同一批次等密度纯化的MMTV中获得高产率的病毒蛋白。首先在1% Triton X-100存在的情况下，用高盐和高pH缓冲液反复提取病毒结构蛋白。通过首先从解离的病毒粒子中用寡聚d（T）柱色谱法纯化较小分子量的病毒蛋白（P14、P12和P10），使柱纯化过程中次要病毒蛋白的损失最小化。然后通过亲和柱和离子交换柱对其他四种主要病毒多肽（GP52、GP36、P28和P23）进行分离。针对除P12和P10之外的所有七种结构蛋白制备了高滴度多克隆抗体。这些抗血清具有单特异性，在放射免疫分析中对其他MMTV蛋白无交叉反应。经过微小修改，该方法也已应用于从几种不同的C型逆转录病毒中纯化其他结构蛋白。
