INTRODUCTION OF SITE-SPECIFIC MUTATIONS INTO THE GENOME OF INFLUENZA-VIRUS
INTRODUCTION OF SITE-SPECIFIC MUTATIONS INTO THE GENOME OF INFLUENZA-VIRUS
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DOI：
10.1073/pnas.87.10.3802
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发表时间：
1990-05-01
影响因子：
11.1
通讯作者：
PALESE, P
中科院分区：
文献类型：
--
作者：
ENAMI, M;LUYTJES, W;PALESE, P
我们通过用源自特定重组 DNA 的 RNA 转染细胞，成功地拯救了传染性流感病毒。与 A/WSN/33 (WSN) 流感病毒的神经氨酸酶 (NA) 基因相对应的 RNA 在体外从质粒 DNA 转录，并在添加纯化的流感病毒 RNA 聚合酶复合物后转染至 MDBK 细胞中。缺乏 WSN NA 基因的辅助病毒的双重感染导致含有 WSN NA 基因的病毒释放。然后，我们通过质粒 DNA 的盒式诱变将五个点突变引入 WSN NA 基因。对获救病毒的序列分析表明，该宝石包含突变质粒中存在的所有五个突变。创造具有位点特异性突变的病毒的能力将允许设计具有特定生物学特性的流感病毒。
We succeeded in rescuing infectious influenza virus by transfecting cells with RNAs derived from specific recombinant DNAs. RNA corresponding to the neuraminidase (NA) gene of influenza A/WSN/33 (WSN) virus was transcribed in vitro from plasmid DNA and, following the addition of purified influenza virus RNA polymerase complex, was transfected into MDBK cells. Superinfection with helper virus lacking the WSN NA gene resulted in the release of virus containing the WSN NA gene. We then introduced five point mutations into the WSN NA gene by cassette mutagenesis of the plasmid DNA. Sequence analysis of the rescued virus revealed that the gemone contained all five mutations present in the mutated plasmid. The ability to create viruses with site-specific mutations will allow the engineering of influenza viruses with defined biological properties.