Levrero M, Jean-Jean O, Balsano C, Will H, Perricaudet M
ER272-IRSC, Villejuif, France.
Virology. 1990 Jan;174(1):299-304. doi: 10.1016/0042-6822(90)90079-7.
All mammalian hepatitis B virus genomes contain an open reading frame X (X-ORF) of unknown function which could encode a protein of 17 kDa. Using a plasmid containing the entire X-ORF preceded by the adenovirus type 2 major late promoter and its tripartite leader sequence efficient expression of the HBV X-gene was achieved. The X protein of 17 kDa was characterized by immunoblotting and immunoprecipitated with an antiserum prepared against a X fusion protein produced in E. coli. By cell fractionation and indirect immunofluorescence the X-protein was found at least in part associated with nuclei. Human cell extracts containing the X protein have been used to screen human sera for anti-HBx antibodies. Such antibodies were detected in sera from patients with active chronic hepatitis with ongoing viral replication. The efficient expression of the HBV X protein obtained will facilitate its functional analysis.
所有哺乳动物乙肝病毒基因组都包含一个功能未知的开放阅读框X（X-ORF），它可编码一种17kDa的蛋白质。利用一个含有完整X-ORF且其前面带有腺病毒2型主要晚期启动子及其三联前导序列的质粒，实现了乙肝病毒X基因的高效表达。通过免疫印迹对17kDa的X蛋白进行了表征，并用针对在大肠杆菌中产生的X融合蛋白制备的抗血清进行了免疫沉淀。通过细胞分级分离和间接免疫荧光发现，X蛋白至少部分与细胞核相关。含有X蛋白的人细胞提取物已用于筛查人血清中的抗-HBx抗体。在正在进行病毒复制的慢性活动性肝炎患者的血清中检测到了此类抗体。所获得的乙肝病毒X蛋白的高效表达将有助于对其进行功能分析。
