Longnecker R, Kieff E
Department of Medicine, Harvard University, Boston, Massachusetts 02115.
J Virol. 1990 May;64(5):2319-26. doi: 10.1128/JVI.64.5.2319-2326.1990.
Recent cDNA cloning and sequencing of two Epstein-Barr virus (EBV)-specific mRNAs from latently infected cultures revealed that these RNAs are encoded across the fused terminal repeats of the viral genome and that they are likely to encode two nearly identical proteins with the same transmembrane domains. The smaller predicted protein (LMP2B) lacks 119 amino-terminal amino acids found in the larger one (LMP2A). To test whether these proteins are expressed in latently infected lymphocytes, antibodies to the LMP2 proteins were derived by immunizing rabbits with TrpE-LMP2A fusion proteins. Affinity-purified LMP2-specific antibodies recognized 54- and 40-kilodalton proteins, corresponding to LMP2A and LMP2B, in immunoblots of rodent fibroblasts stably transfected with eucaryotic expression plasmids containing either the LMP2A or LMP2B cDNA. Similar-size proteins were also identified in immunoblots of latently infected lymphocytes. LMP2A localized to membranes in cellular fractionation studies. In immunofluorescent studies, LMP2 localized in the plasma membrane of EBV-infected lymphocytes, with the majority of reactivity confined to the region of the LMP1 patch. This reactivity was detected in almost all lymphoblastoid cells latently infected with EBV.
最近,对来自潜伏感染培养物的两种爱泼斯坦-巴尔病毒(EBV)特异性mRNA进行的cDNA克隆和测序显示,这些RNA是跨越病毒基因组的融合末端重复序列编码的,并且它们可能编码两种具有相同跨膜结构域的几乎相同的蛋白质。预测的较小蛋白质(LMP2B)缺少较大蛋白质(LMP2A)中发现的119个氨基末端氨基酸。为了测试这些蛋白质是否在潜伏感染的淋巴细胞中表达,通过用TrpE-LMP2A融合蛋白免疫兔子获得了针对LMP2蛋白的抗体。亲和纯化的LMP2特异性抗体在稳定转染了含有LMP2A或LMP2B cDNA的真核表达质粒的啮齿动物成纤维细胞的免疫印迹中识别出54千道尔顿和40千道尔顿的蛋白质,分别对应于LMP2A和LMP2B。在潜伏感染淋巴细胞的免疫印迹中也鉴定出了类似大小的蛋白质。在细胞分级分离研究中,LMP2A定位于细胞膜。在免疫荧光研究中,LMP2定位于EBV感染淋巴细胞的质膜,大部分反应局限于LMP1斑块区域。在几乎所有潜伏感染EBV的淋巴母细胞中都检测到了这种反应。