完整、具有感染性的爱泼斯坦-巴尔病毒从原核细胞到人类细胞的传播与复苏。
Propagation and recovery of intact, infectious Epstein-Barr virus from prokaryotic to human cells.
作者信息
Delecluse H J, Hilsendegen T, Pich D, Zeidler R, Hammerschmidt W
机构信息
National Research Center for Environment and Health, Institut für Klinische Molekularbiologie und Tumorgenetik, Marchioninistr. 25, D-81377 Munich, Germany.
出版信息
Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8245-50. doi: 10.1073/pnas.95.14.8245.
With current techniques, genetic alterations of herpesviruses are difficult to perform, mostly because of the large size of their genomes. To solve this problem, we have designed a system that allows the cloning of any gamma-herpesvirus in Escherichia coli onto an F factor-derived plasmid. Immortalized B cell lines were readily established with recombinant Epstein-Barr virus (EBV), demonstrating that the F factor-cloned EBV genome has all the characteristics of wild-type EBV. Because any genetic modification is possible in E. coli, this experimental approach opens the way to the genetic analysis of all EBV functions. Moreover, it is now feasible to generate attenuated EBV strains in vitro such that vaccine strains can be designed. Because we incorporated the genes for hygromycin resistance and green fluorescent protein onto the E. coli cloned EBV genome, the still open question of the EBV target cells other than B lymphocytes will be addressed.
利用当前技术，疱疹病毒的基因改造很难进行，主要原因是其基因组规模庞大。为解决这一问题，我们设计了一个系统，可将任何γ-疱疹病毒在大肠杆菌中克隆到一个F因子衍生质粒上。利用重组爱泼斯坦-巴尔病毒（EBV）能轻易建立永生化B细胞系，这表明F因子克隆的EBV基因组具有野生型EBV的所有特征。由于在大肠杆菌中可以进行任何基因改造，这种实验方法为所有EBV功能的基因分析开辟了道路。此外，现在在体外产生减毒EBV毒株从而设计疫苗毒株是可行的。因为我们将潮霉素抗性基因和绿色荧光蛋白基因整合到大肠杆菌克隆的EBV基因组上，除B淋巴细胞外的EBV靶细胞这一仍未解决的问题将得到解决。
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