IMMORTALIZATION OF HUMAN B-LYMPHOCYTES BY A PLASMID CONTAINING 71 KILOBASE PAIRS OF EPSTEIN-BARR-VIRUS DNA
IMMORTALIZATION OF HUMAN B-LYMPHOCYTES BY A PLASMID CONTAINING 71 KILOBASE PAIRS OF EPSTEIN-BARR-VIRUS DNA
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DOI：
10.1128/jvi.69.1.231-238.1995
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发表时间：
1995-01-01
影响因子：
5.4
通讯作者：
HAMMERSCHMIDT, W
中科院分区：
文献类型：
--
作者：
KEMPKES, B;PICH, D;HAMMERSCHMIDT, W
我们已经组装了EB病毒（EBV）的衍生物，其中包括71 kbp的非连续DNA序列克隆到原核F-因子质粒。当将这些微型EBV引入含有EBV的淋巴母细胞样细胞中时，可以通过内源性辅助病毒包装。发现一种这样的迷你EB病毒从其EBNA 3a开放阅读框中缺失了一个C残基。当被包装时，这种微型EBV仅与互补辅助病毒结合启动受感染的原代人B淋巴细胞的增殖。然而，感染细胞的增殖或者单独由含有突变的EBNA 3a开放阅读框的迷你EBV维持，或者单独由其衍生物维持，其中EBNA 3a开放阅读框已经通过与辅助病毒重组而治愈其损伤。当单独包装时，具有野生型EBNA 3a开放阅读框的迷你EBV在感染原代人B淋巴细胞后可以启动和维持增殖。这些发现鉴定了41%的EBV DNA，其足以使原代人B淋巴细胞永生化，并提供了区分病毒对细胞增殖的启动或维持或两者的贡献的测定。他们还将EBNA 3a鉴定为转化基因，其主要有助于细胞增殖的启动。
We have assembled derivatives of Epstein-Barr Virus (EBV) that include 71 kbp of noncontiguous DNA sequences cloned into a prokaryotic F-factor plasmid. These mini-EBVs, when introduced into an EBV-containing lymphoblastoid cell, can be packaged by the endogenous helper virus. One such mini-EBV was found to have a single C residue deleted from its EBNA3a open reading frame. When packaged, this mini-EBV initiates proliferation of infected primary human B lymphocytes only in conjunction with a complementing helper virus. Proliferation of the infected cells, however, was maintained either alone by the mini-EBV containing the mutated EBNA3a open reading frame or alone by its derivative in which the EBNA3a open reading frame had been healed of its lesion by recombination with the helper virus. The mini-EBV with a wild-type EBNA3a open reading frame when packaged alone can both initiate and maintain proliferation upon infection of primary human B lymphocytes. These findings identify 41% of EBV DNA which is sufficient to immortalize primary human B lymphocytes and provide an assay to distinguish virus contributions to initiation or maintenance of cell proliferation or both. They also identify EBNA3a as a transforming gene, which contributes primarily to the initiation of cell proliferation.