Construction and characterization of an infectious DNA clone and of mutants of simian immunodeficiency virus isolated from the African green monkey
Construction and characterization of an infectious DNA clone and of mutants of simian immunodeficiency virus isolated from the African green monkey
复制标题
从非洲绿猴中分离的猿猴免疫缺陷病毒感染性 DNA 克隆和突变体的构建和表征
DOI：
10.1128/jvi.64.1.307-312.1990
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发表时间：
1990
影响因子：
5.4
通讯作者：
Akio Adachi
中科院分区：
文献类型：
--
作者：
R. Shibata;Tomoyuki Miura;Masanori Hayami;H. Sakai;K. Ogawa;T. Kiyomasu;A. Ishimoto;Akio Adachi
我们从一只非洲绿色猴中构建了猴免疫缺陷病毒的全长分子克隆。在转染后，该克隆指导产生致细胞病变的病毒颗粒，并感染人CD4+白血病细胞系。通过重组DNA技术将突变引入到迄今为止鉴定的来自非洲绿色猴的猴免疫缺陷病毒的8个开放阅读框中。突变病毒的表型，即，通过转染和感染实验检查感染性、致细胞病变性、由长末端重复序列控制的基因表达的反式激活以及病毒RNA和蛋白质合成。三个结构（gag，pol和env）和两个调节（达特和rev）基因突变体没有感染性，而vif，vpx和nef的感染性和突变病毒是高度致细胞病变。在瞬时转染试验中，rev突变体主要产生小的mRNA种类，没有检测到病毒蛋白和颗粒。达特突变体的反式激活潜力比野生型DNA低约10倍，产生少量病毒。
We constructed a full-length molecular clone of simian immunodeficiency virus from an African green monkey. Upon transfection, this clone directed the production of virus particles cytopathic and infectious to human CD4+ leukemia cell lines. Mutations were introduced by recombinant DNA techniques into eight open reading frames of simian immunodeficiency virus from the African green monkey thus far identified. The phenotypes of mutant viruses, i.e., infectivity, cytopathogenicity, transactivation of gene expression controlled by a long terminal repeat, and viral RNA and protein syntheses, were examined by transfection and infection experiments. Three structural (gag, pol, and env) and two regulatory (tat and rev) gene mutants were not infectious, whereas vif, vpx, and nef were dispensable for infectivity and mutant viruses were highly cytopathic. In transient transfection assays, a rev mutant produced mainly small mRNA species and no detectable virus protein and particles. The transactivation potential of a tat mutant was about 10-fold less than that of wild-type DNA, generating small amounts of virus.