Portugal Raquel S, Bauer Anja, Keil Guenther M
Institut für molekulare Virologie und Zellbiologie, Friedrich-Loeffler-Institut, Südufer 10, Greifswald, Insel Riems 17493, Germany.
Institut für molekulare Virologie und Zellbiologie, Friedrich-Loeffler-Institut, Südufer 10, Greifswald, Insel Riems 17493, Germany.
Virology. 2017 Aug;508:70-80. doi: 10.1016/j.virol.2017.05.007. Epub 2017 May 11.
African swine fever virus threatens pig production worldwide due to the lack of vaccines, for which generation of both deletion and insertion mutants is considered. For development of the latter, operational ASFV promoters of different temporal regulation and strengths are desirable. We therefore compared the capacities of putative promoter sequences from p72, CD2v, p30, viral DNA polymerase and U104L genes to mediate expression of luciferase from transfected plasmids after activation in trans, or p30-, DNA polymerase- and U104L promoters in cis, using respective ASFV recombinants. We identified sequences with promoter activities upstream the viral ORFs, and showed that they differ in both their expression intensity regulating properties and in their temporal regulation. In summary, p30 and DNA polymerase promoters are recommended for high level early regulated transgene expression. For late expression, the p72, CD2v and U104L promoter are suitable. The latter however, only if low level transgene expression is aimed.
由于缺乏疫苗，非洲猪瘟病毒对全球养猪业构成威胁，因此人们考虑培育缺失和插入突变体。为了培育后者，需要不同时间调控和强度的可用非洲猪瘟病毒启动子。因此，我们比较了来自p72、CD2v、p30、病毒DNA聚合酶和U104L基因的假定启动子序列在反式激活后介导转染质粒中荧光素酶表达的能力，或者使用各自的非洲猪瘟病毒重组体比较了顺式中p30、DNA聚合酶和U104L启动子的能力。我们在病毒开放阅读框上游鉴定出具有启动子活性的序列，并表明它们在表达强度调控特性和时间调控方面都有所不同。总之，推荐使用p30和DNA聚合酶启动子进行高水平的早期调控转基因表达。对于晚期表达，p72、CD2v和U104L启动子是合适的。然而，只有在目标是低水平转基因表达的情况下才使用后者。
