Behzadian Farida, Sabahi Farzaneh, Sadeghizadeh Majid, Maghsoudi Nader, Fotouhi Fatemeh, Karimi Mohsen
Department of Virology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Intervirology. 2007;50(6):387-93. doi: 10.1159/000110650. Epub 2007 Oct 31.
There is no cell line susceptible to hepatitis D virus (HDV) infection and capable of stable replication of its genome. Different genetic-based approaches have been introduced to initiate HDV replication events so far.
In order to construct a replicative model for HDV made from a unique genome sequence, two monomeric units of HDV full-length cDNA were joined together through a four-step cloning scheme. The resulting vector (pcDNA3.1-D2) containing two tandem repeats of HDV cDNA under CMV promoter control was then used in transfection experiments into COS7 and HuH7 cell lines.
HDV replication markers including expression of hepatitis delta antigen (HDAg), the only HDV-specific antigen, and synthesis of antigenomic RNA were shown in both transfected cell lines, indicating initiation of HDV genome replication.
Our results suggested that pcDNA3.1-D2, a vector containing a cDNA dimer of the HDV genome, originated from a unique full-length HDV molecule that is capable of replicating in cultured cells. This vector can be used conveniently for transfection experiments to study HDV molecular biology.
目前尚无对丁型肝炎病毒（HDV）感染敏感且能稳定复制其基因组的细胞系。迄今为止，已引入了不同的基于基因的方法来启动HDV复制事件。
为构建一个由独特基因组序列构成的HDV复制模型，通过四步克隆方案将HDV全长cDNA的两个单体单元连接在一起。然后将所得载体（pcDNA3.1-D2），其在CMV启动子控制下含有HDV cDNA的两个串联重复序列，用于转染COS7和HuH7细胞系的实验。
在两个转染细胞系中均显示出HDV复制标志物，包括丁型肝炎抗原（HDAg）的表达（唯一的HDV特异性抗原）以及反基因组RNA的合成，表明HDV基因组复制开始。
我们的结果表明，pcDNA3.1-D2，一种含有HDV基因组cDNA二聚体的载体，源自一个能够在培养细胞中复制的独特全长HDV分子。该载体可方便地用于转染实验以研究HDV分子生物学。
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