3 INTERGENIC REGIONS OF CORONAVIRUS MOUSE HEPATITIS-VIRUS STRAIN-A59 GENOME RNA CONTAIN A COMMON NUCLEOTIDE-SEQUENCE THAT IS HOMOLOGOUS TO THE 3' END OF THE VIRAL MESSENGER-RNA LEADER SEQUENCE
3 INTERGENIC REGIONS OF CORONAVIRUS MOUSE HEPATITIS-VIRUS STRAIN-A59 GENOME RNA CONTAIN A COMMON NUCLEOTIDE-SEQUENCE THAT IS HOMOLOGOUS TO THE 3' END OF THE VIRAL MESSENGER-RNA LEADER SEQUENCE
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DOI：
10.1128/jvi.53.3.834-840.1985
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发表时间：
1985-01-01
影响因子：
5.4
通讯作者：
WEISS, SR
中科院分区：
文献类型：
--
作者：
BUDZILOWICZ, CJ;WILCZYNSKI, SP;WEISS, SR
构建了代表冠状病毒小鼠肝炎病毒株A59基因组RNA不同部分的c[互补]DNA克隆。以小牛胸腺DNA的寡聚体（oligo（dT）12-18）或随机寡聚体（random oligomers）为引物，通过基因组RNA的转录合成cDNA。将这些cDNA转化为双链DNA，并通过标准技术克隆到pBR 322中。通过与来自小鼠肝炎病毒株A59感染的小鼠成纤维细胞的细胞内RNA的北方印迹杂交，将所得克隆的病毒DNA片段定位到病毒基因。这些cDNA克隆定位于7个病毒基因中的6个。克隆g344，1.8个内切酶，是最大的，包括基因5（编码非结构蛋白）和基因6（编码E1病毒糖蛋白）以及基因5、6和7之前的基因间区域。部分克隆DNA的测序表明，这3个基因间区域包含一个共同的11个核苷酸序列。该序列与病毒mRNA前导序列的3“末端具有同源性。因此，这种共同的基因间序列可能含有前导RNA的结合位点，该前导RNA在每个基因的开始处与负链病毒RNA杂交以引发mRNA合成。前导序列与其假定结合位点之间的不同程度的同源性可能影响各种病毒mRNA的不同转录速率。
c[complementary]DNA clones that represent various portions of the coronavirus mouse hepatitis virus strain A59 genome RNA were constructed. cDNA were synthesized by transcription of genome RNA by using either oligo(dT)12-18 or random oligomers of calf thymus DNA as primers. These cDNA were converted into double-stranded DNA and cloned into pBR322 by standard techniques. The resulting cloned viral DNA fragments were mapped to viral genes by hybridization with Northern blots of intracellular RNA from mouse hepatitis virus strain A59-infected mouse fibroblast cells. These cDNA clones map in 6 of the 7 viral genes. Clone g344, 1.8 kilobases, is the largest and encompasses gene 5 (which encodes a nonstructural protein) and gene 6 (which encodes the E1 viral glycoprotein) as well as the intergenic regions preceding genes 5, 6 and 7. Sequencing of parts of this cloned DNA show that these 3 intergenic regions contain a common 11-nucleotide sequence. This sequence shares homology with the 3'' end of the viral mRNA leader sequence. Thus, this common intergenic sequence may contain a binding site for a leader RNA that hybridizes to negative-strand viral RNA at the beginning of each gene to prime mRNA synthesis. The different degrees of homology between the leader and its putative binding site may influence the differential rates of transcription of the various viral mRNA.