Yogo Y, Hirano N, Hino S, Shibuta H, Matumoto M
J Biochem. 1977 Oct;82(4):1103-8. doi: 10.1093/oxfordjournals.jbchem.a131782.
Mouse hepatitis (MH) virus was grown in SR-CDF1-DBT, a mouse cell line, and purified by ammonium sulfate precipitation and by density gradient centrifugation. Extraction of RNA from purified virions with 1% SDS and sedimentation analysis of the RNA revealed a major 50S component and two minor components. Treatment of virions with phenol/chloroform also produced the 50S component, although its yield was lower. MH virion RNA can bind to a poly(U)-fiberglass filter, indicating that MH virion RNA contains poly(A). A poly(A)-like fragment was isolated by digestion with ribonuclease A [EC 3.1.4.22] and T1 [EC 3.1.4.8] and by DEAE-Sephadex column chromatography. Analysis of the fragment for base composition showed it to be an adenine-rich material. Its chain length was about 90 nucleotides, as determined by ion-exchange chromatography and gel electrophoresis.
小鼠肝炎(MH)病毒在小鼠细胞系SR-CDF1-DBT中培养,并通过硫酸铵沉淀和密度梯度离心进行纯化。用1%十二烷基硫酸钠(SDS)从纯化的病毒粒子中提取RNA,并对RNA进行沉降分析,结果显示有一个主要的50S组分和两个次要组分。用苯酚/氯仿处理病毒粒子也能产生50S组分,尽管其产量较低。MH病毒粒子RNA能与聚(U)-玻璃纤维滤膜结合,这表明MH病毒粒子RNA含有聚(A)。通过用核糖核酸酶A [EC 3.1.4.22]和T1 [EC 3.1.4.8]消化以及通过二乙氨基乙基葡聚糖(DEAE-Sephadex)柱色谱法分离出一个类似聚(A)的片段。对该片段的碱基组成进行分析表明它是一种富含腺嘌呤的物质。通过离子交换色谱法和凝胶电泳测定,其链长约为90个核苷酸。