Analysis and gene assignment of mRNAs of a paramyxovirus, simian virus 5.
Analysis and gene assignment of mRNAs of a paramyxovirus, simian virus 5.
复制标题
副粘病毒、猿猴病毒 5 的 mRNA 分析和基因分配。
DOI：
10.1016/0042-6822(84)90354-4
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发表时间：
1984
期刊：
影响因子：
3.7
通讯作者：
Lamb,RA
中科院分区：
文献类型：
--
作者：
Paterson,RG;Harris,TJ;Lamb,RA
当用放线菌素D处理宿主细胞时，在感染的CV-1细胞中由副粘病毒SV 5合成的多肽很容易被鉴定。用特异性抗体免疫沉淀法鉴定了在衣霉素存在下感染细胞中合成的HN和Fo的非糖基化形式以及体外合成的HN和Fo。在甲基汞琼脂糖凝胶上分离SV 5特异性的含poly（A）的RNA，并在体外翻译片段，表明病毒多肽是从单个mRNA翻译的，除了P（MrP 44 K）和非结构多肽V（MrP 24 K）之外，它们的mRNA不能分离。合成SV 5特异性mRNA的cDNA拷贝并克隆到质粒pBR 322中。通过SV 5 mRNA的杂交捕获和杂交选择翻译鉴定NP、P + V、M、F和HN的克隆。胰蛋白酶肽映射的多肽P和V表明，肽V的一个子集的P.杂交的cDNA探针感染的细胞mRNA上分离的琼脂糖凝胶允许识别的NP，P + V，M，F，和HN mRNA和推定的多顺反子mRNA。这些多顺反子mRNA的大小和序列同源性用于推导SV 5 50 S基因组RNA上可能的基因顺序。
Polypeptides synthesized by the paramyxovirus SV5 in infected CV-1 cells were readily identified when the host cell was treated with actinomycin D. The unglycosylated forms of HN and Fosynthesized in infected cells in the presence of tunicamycin and HN and Fosynthesizedin vitrowere identified by immunoprecipitation with specific antibodies. Separation of SV5-specific poly(A)-containing RNAs on methyl-mercury agarose gels andin vitrotranslation of fractions, indicated that the viral polypeptides were translated from individual mRNAs except P (Mr∼ 44K) and the nonstructural polypeptide V (Mr∼ 24K) for which the mRNAs could not be separated. CDNA copies of SV5-specific mRNAs were synthesized and cloned in plasmid pBR322. Clones to NP, P + V, M, F, and HN were identified by hybrid-arrest and hybrid-selection translation of SV5 mRNAs. Tryptic peptide mapping of polypeptides P and V indicated that the peptides of V were a subset of those of P. Hybridization of cDNA probes to infected cell mRNAs separated on agarose gels permitted identification of the NP, P + V, M, F, and HN mRNAs and presumptive polycistronic mRNAs. The sizes and sequence homologies of these polycistronic mRNAs were used to derive a likely gene order on the SV5 50 S genome RNA.