Alt M, Fleckenstein B, Grassmann R
Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, F.R.G.
Gene. 1991 Jun 30;102(2):265-9. doi: 10.1016/0378-1119(91)90088-s.
The genome of Herpesvirus saimiri, a lymphotropic virus of non-human primates, was used to develop a vector system for transducing foreign genes into primary human T-cells and T-lymphoid cell lines. Recombinant viruses were obtained by homologous recombination of the viral genome with linearized plasmid DNA. The plasmid used contained a fragment of virion DNA, a hygromycin-B-resistance marker (HyR), and a multiple cloning site for the insertion of additional expression cassettes. The resulting recombinants were efficiently enriched and were plaque-purified. The virus mediating HyR and a H. saimiri strain carrying the Geneticin-resistance marker were used to infect the human T-lymphoid cell line Jurkat. Lymphocytes with a double-resistant phenotype were shown to contain the two different H. saimiri recombinants persisting as episomes at high multiplicity. The H. saimiri vector system will be suitable to study cooperating regulatory genes in T-lymphocytes.
赛氏疱疹病毒是一种非人类灵长类动物的嗜淋巴细胞病毒，其基因组被用于开发一种载体系统，用于将外源基因转导到原代人T细胞和T淋巴细胞系中。通过病毒基因组与线性化质粒DNA的同源重组获得重组病毒。所使用的质粒包含一段病毒粒子DNA片段、一个潮霉素B抗性标记（HyR）以及一个用于插入额外表达盒的多克隆位点。所得重组体被有效地富集并进行了噬斑纯化。介导HyR的病毒和携带遗传霉素抗性标记的赛氏疱疹病毒菌株被用于感染人T淋巴细胞系Jurkat。具有双抗性表型的淋巴细胞被证明含有两种不同的赛氏疱疹病毒重组体，它们以高拷贝数作为附加体持续存在。赛氏疱疹病毒载体系统将适用于研究T淋巴细胞中协同调控基因。
