Hall M R, Meinke W, Goldstein D A
J Virol. 1973 Oct;12(4):901-8. doi: 10.1128/JVI.12.4.901-908.1973.
Procedures for isolating nucleoprotein complexes containing replicating polyoma DNA from infected mouse cells were used to prepare short-lived nucleoprotein complexes (r-SV40 complexes) containing replicating simian virus 40 (SV40) DNA from infected monkey cells. Like the polyoma complexes, r-SV40 complexes were only partially released from nuclei by cell lysis but could be extracted from nuclei by prolonged treatment with solutions containing Triton X-100. r-SV40 complexes sedimented faster than complexes containing SV40 supercoiled DNA (SV40 complex) in sucrose gradients, and both types of SV40 nucleoprotein complexes sedimented ahead of polyoma complexes containing supercoiled polyoma DNA (py complex). The sedimentation rates of py complex and SV40 complex were 56 and 61S, respectively, based on the sedimentation rate of the mouse large ribosomal subunit as a marker. r-SV40 complexes sedimented as multiple peaks between 56 and 75S. Sedimentation and buoyant density measurements indicated that protein is bound to all forms of SV40 DNA at about the same ratio of protein to DNA (1-2/1) as was reported for polyoma nucleoproteins.
使用从感染的小鼠细胞中分离含复制多瘤病毒DNA的核蛋白复合物的方法,从感染的猴细胞中制备了含复制猿猴病毒40(SV40)DNA的短寿命核蛋白复合物(r-SV40复合物)。与多瘤病毒复合物一样,r-SV40复合物仅通过细胞裂解从细胞核中部分释放,但可通过用含Triton X-100的溶液长时间处理从细胞核中提取。在蔗糖梯度中,r-SV40复合物的沉降速度比含SV40超螺旋DNA的复合物(SV40复合物)快,并且两种类型的SV40核蛋白复合物都比含超螺旋多瘤病毒DNA的多瘤病毒复合物(py复合物)沉降得快。以小鼠大核糖体亚基的沉降速度为标记,py复合物和SV40复合物的沉降速度分别为56和61S。r-SV40复合物以56至75S之间的多个峰沉降。沉降和浮力密度测量表明,蛋白质与所有形式的SV40 DNA结合的蛋白质与DNA的比例(1-2/1)与多瘤病毒核蛋白报道的比例大致相同。