ORGANIZATION OF NONSTRUCTURAL GENES OF THE AUTONOMOUS PARVOVIRUS MINUTE VIRUS OF MICE
ORGANIZATION OF NONSTRUCTURAL GENES OF THE AUTONOMOUS PARVOVIRUS MINUTE VIRUS OF MICE
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DOI：
10.1128/jvi.58.3.724-732.1986
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发表时间：
1986-06-01
影响因子：
5.4
通讯作者：
TATTERSALL, P
中科院分区：
文献类型：
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作者：
COTMORE, SF;TATTERSALL, P
将小鼠自主细小病毒（autonomous parvovirus minute virus， MVM）基因组的开放阅读框（open reading frame， ORF）片段克隆到原核表达载体中，分离出含有MVM特异性氨基酸序列的细菌融合蛋白。针对这些蛋白产生的抗体用于免疫沉淀在兔网织细胞裂解物翻译系统中体外合成的病毒蛋白，该翻译系统由从MVM和许多不同细小病毒感染的细胞中分离的mRNA编程。这些研究表明：83千道尔顿的非结构蛋白NS-1和25千道尔顿的非结构蛋白NS-2具有共同的氨基末端序列，该序列由病毒基因组中位于核苷酸225和534之间的单个ORF编码；位于核苷酸1110和1638之间的ORF仅在NS-1蛋白中表达；在核苷酸2075和2291之间的小备选ORF中编码的序列仅在NS-2中表达。这些数据证实了NS-1是4.8千碱基R1病毒转录本的产物，并证明NS-2是由3.3千碱基R2转录本合成的，后者产生于病毒基因组图单元4的左侧启动子。针对MVM融合蛋白的抗体也与病毒H-1和LuIII编码的类似蛋白发生交叉反应，但尽管针对NS-1羧基末端一半的抗体与CPV[犬细小病毒]中的类似蛋白发生交叉反应，我们无法证明该病毒编码的NS-2蛋白。
Regions of open reading frame (ORF) from the genome of the autonomous parvovirus minute virus of mice (MVM) were cloned into a procaryotic expression vector, and bacterial fusion protens containing MVM-specific aminoacid sequences were isolated. Antibodies raised against these proteins were used to immunoprecipitate viral proteinssynthesized in vitro in a rabbit reticulocyte lysate translation system programmed with mRNA isolated from the cells infected with MVM and a number of different parvoviruses. These studies demonstrated that: the 83-kilodalton nonstructural protein NS-1 and the 25-kilodalton nonstructural protein NS-2 have a common amino-terminal sequence which is encoded by the single ORF located between nucleotides 225 and 534 in the viral genome; the ORF located between nucleotides 1110 and 1638 is only expressed in the NS-1 protein; and the sequence encoded in a small alternative ORF between nucleotides 2075 and 2291 is expressed exclusively in NS-2. These data confirm that NS-1 is the product of the 4.8-kilobase R1 viral transcript and demonstrate that NS-2 is synthesized from the 3.3-kilobase R2 transcript which arises from the left-hand promoter at map unit 4 on the viral genome. Antibodies against the MVM fusion proteins also cross-reacted with similar proteins encoded by the viruses H-1 and LuIII, but although antibodies against the carboxyl-terminal half of the NS-1 cross-reacted with a similar protein in CPV [canine parvovirus], we were unable to demonstrate an NS-2 protein encoded by this virus.