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与小鼠乳腺肿瘤病毒长末端重复序列编码的肽具有抗原相关性的蛋白质与胞质内A颗粒有关。

Proteins antigenically related to peptides encoded by the mouse mammary tumour virus long terminal repeat sequence are associated with intracytoplasmic A particles.

作者信息

Smith G H, Young L J, Benjamini F, Medina D, Cardiff R D

出版信息

J Gen Virol. 1987 Feb;68 ( Pt 2):473-86. doi: 10.1099/0022-1317-68-2-473.

Abstract

Intracytoplasmic A particles (CAP), previously identified as cytoplasmic nucleocapsid precursors to mouse mammary tumour virus (MMTV), reacted strongly in immunodiffusion tests with polyclonal antibodies raised against synthetic oligopeptides derived from the open reading frame (ORF) in the long terminal repeat (LTR) of MMTV. In Western blots, several CAP proteins (p80, p72-68, p36, p32, p18-12) were reactive with polyclonal antibodies raised against three separate LTR ORF synthetic peptides. Disrupted MMTV virions did not react with the anti-LTR ORF peptides suggesting that ORF proteins were excluded from mature virions during maturation. Serial dilution of anti-LTR ORF antibody demonstrated that the most reactive CAP proteins in Western blots migrated as a doublet band with estimated molecular weights of 68,000 and 72,000. Reactivity of anti-LTR ORF serum with these and other CAP proteins was removed upon preincubation with free synthetic peptide. Absorption with LTR synthetic peptides did not affect the reactivity of antibodies directed against MMTV gag proteins with similarly sized CAP polyproteins. LTR ORF-related proteins with molecular weights similar to those associated with CAP were also detectable in Western blots of total cytoplasmic extracts of MMTV-infected mammary tumour cells.

摘要

胞质内A颗粒(CAP),先前被鉴定为小鼠乳腺肿瘤病毒(MMTV)的细胞质核衣壳前体,在免疫扩散试验中与针对源自MMTV长末端重复序列(LTR)开放阅读框(ORF)的合成寡肽产生的多克隆抗体发生强烈反应。在蛋白质免疫印迹中,几种CAP蛋白(p80、p72 - 68、p36、p32、p18 - 12)与针对三种不同LTR ORF合成肽产生的多克隆抗体发生反应。破坏的MMTV病毒粒子不与抗LTR ORF肽反应,这表明在成熟过程中ORF蛋白被排除在成熟病毒粒子之外。抗LTR ORF抗体的系列稀释表明,在蛋白质免疫印迹中反应性最强的CAP蛋白以双重条带形式迁移,估计分子量为68,000和72,000。抗LTR ORF血清与这些及其他CAP蛋白的反应性在与游离合成肽预孵育后消失。用LTR合成肽吸收并不影响针对MMTV gag蛋白的抗体与大小相似的CAP多蛋白的反应性。在MMTV感染的乳腺肿瘤细胞总细胞质提取物的蛋白质免疫印迹中也可检测到分子量与CAP相关的LTR ORF相关蛋白。

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