Yoo B J, Selby M J, Choe J, Suh B S, Choi S H, Joh J S, Nuovo G J, Lee H S, Houghton M, Han J H
Chiron Corporation, Emeryville, California 94608.
J Virol. 1995 Jan;69(1):32-8. doi: 10.1128/JVI.69.1.32-38.1995.
T7 RNA polymerase transcripts of a putative full-length cDNA clone of hepatitis C virus type 1 (HCV-1) were used to transfect a differentiated human hepatoma cell line, Huh7. The transfected genome replicated in cells, as evidenced by the appearance of progeny HCV RNA, detection of negative-strand viral RNA, and incorporation of [3H]uridine into the viral genome. Incubation of naive Huh7 cells with conditioned medium from transfected cells resulted in a new HCV infection, suggesting the production of biologically active virus in the inoculum. Maintenance of the transfected cells under serum-free culture conditions resulted in the selection of persistently infected cells which displayed a distinctive cellular morphology. This is the first demonstration that HCV RNA produced from cloned HCV cDNA is infectious and replication competent. This approach should provide a valuable system for studying HCV replication, persistence, and pathogenicity.
利用1型丙型肝炎病毒（HCV-1）推定全长cDNA克隆的T7 RNA聚合酶转录本转染分化的人肝癌细胞系Huh7。转染的基因组在细胞中复制，子代HCV RNA的出现、负链病毒RNA的检测以及[3H]尿苷掺入病毒基因组都证明了这一点。用来自转染细胞的条件培养基培养未处理的Huh7细胞导致新的HCV感染，这表明接种物中产生了具有生物活性的病毒。在无血清培养条件下维持转染细胞导致选择出具有独特细胞形态的持续感染细胞。这是首次证明从克隆的HCV cDNA产生的HCV RNA具有感染性且具备复制能力。这种方法应为研究HCV复制、持续性和致病性提供一个有价值的系统。
