Directed Evolution of Adenoviruses.

Directed Evolution of Adenoviruses.
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腺病毒的定向进化。

DOI:
10.1007/978-1-4939-3271-9_13
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发表时间:
2016
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Smith,JasonG
Smith,JasonG
中科院分区:
--
文献类型:
--
作者:
Smith,JasonG

文献摘要

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面对选择压力,病毒在细胞培养中进化的能力是阐明合成或天然抗病毒药物的分子机制、扩大向性或改变毒力的宝贵方法。最近,描述了降低复制保真度的人腺病毒聚合酶的突变,并且我们已经将这些突变之一掺入基于条件复制人腺病毒血清型5(HAdV-5)的载体的pol基因中(Uil等人,Nucleic Acids Res 39:e30,2011; Myers等人,J Virol 87:6047-6050,2013)。在这里,我们描述的方法来发展这种病毒(HAdV-5.polF421Y)的选择压力下,从抗病毒药物,以确定其作用机制。
The ability to evolve viruses in cell culture in the face of selective pressure is an invaluable method to elucidate the molecular mechanisms of synthetic or natural antivirals, expand tropism, or alter virulence. Recently, mutations to the human adenovirus polymerase that reduce replicative fidelity were described, and we have incorporated one of these mutations into thepolgene of a conditionally replicating human adenovirus serotype 5 (HAdV-5)-based vector (Uil et al., Nucleic Acids Res 39:e30, 2011; Myers et al., J Virol 87:6047–6050, 2013). Here, we describe methods to evolve this virus (HAdV-5.polF421Y) under selective pressure from antivirals to identify their mechanisms of action.