Yaginuma K, Shirakata Y, Kobayashi M, Koike K
Proc Natl Acad Sci U S A. 1987 May;84(9):2678-82. doi: 10.1073/pnas.84.9.2678.
An in vitro system for the production of hepatitis B virus (HBV) particles was established by the transient expression of transfected HBV DNA using a human hepatocellular carcinoma cell, HuH-7, as a recipient. The 3.6- and 2.2-kilobase transcripts observed were similar to those in virus-infected liver cells. Both transcripts revealed the microheterogeneity of their 5' ends. The formation of virus-related particles subsequent to the RNA transcription was demonstrated. The core particles observed in the cytoplasm and the virus particles secreted in the culture medium contained the replicative intermediates of HBV DNA and banded at densities of 1.35-1.36 g/cm3 and 1.22-1.24 g/cm3, respectively. Furthermore, the in vitro mutagenesis of the template HBV DNA demonstrated that the P gene as well as the C gene products were essential for the production of HBV particles.
通过使用人肝癌细胞HuH-7作为受体，对转染的乙肝病毒（HBV）DNA进行瞬时表达，建立了一种体外生产HBV颗粒的系统。观察到的3.6千碱基和2.2千碱基转录本与病毒感染的肝细胞中的转录本相似。两种转录本均显示出其5'端的微异质性。证明了RNA转录后病毒相关颗粒的形成。在细胞质中观察到的核心颗粒和培养基中分泌的病毒颗粒含有HBV DNA的复制中间体，其密度分别为1.35 - 1.36 g/cm³和1.22 - 1.24 g/cm³。此外，对模板HBV DNA的体外诱变表明，P基因以及C基因产物对于HBV颗粒的产生至关重要。
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