Barr S M, Keck K, Aposhian H V
Department of Cellular and Developmental Biology, University of Arizona, Tucson, Arizona 85721, USA.
Virology. 1979 Jul 30;96(2):656-9. doi: 10.1016/0042-6822(79)90124-7.
A polyoma-like particle (PLP) is formed when polyoma DNA and purified empty capsids are incubated in a cell-free system. The DNA of this new particle is protected against the action of pancreatic DNase. The density of the purified PLP in CsCl is 1.32 g/cm3, which is intermediate between that of polyoma virions (1.34 g/cm3) and empty capsids (1.29 g/cm3). Purified PLP sediments at 190 S in sucrose and is stable in solutions of high ionic strength. When the DNA is extracted from PLP by the use of detergent and phenol, it is found to be doublestranded with a molecular weight of approximately 1.1 x 10(6). The particles are stable in CsCl at 4 degrees for at least 5 months. Electron micrographs indicate that highly purified PLPs stained with 2% PTA have the same appearance as polyoma capsids. Neither aggregates nor complexes bound by loose ionic bonds appear reasonable to explain these results. The evidence indicates that the DNA of this new polyoma-like particle, made under cell-free conditions, is protected by the capsid.
当多瘤病毒DNA与纯化的空衣壳在无细胞系统中孵育时,会形成一种多瘤样颗粒(PLP)。这种新颗粒的DNA可免受胰脱氧核糖核酸酶的作用。纯化后的PLP在CsCl中的密度为1.32 g/cm³,介于多瘤病毒粒子(1.34 g/cm³)和空衣壳(1.29 g/cm³)之间。纯化后的PLP在蔗糖中以190 S沉降,且在高离子强度溶液中稳定。当使用去污剂和苯酚从PLP中提取DNA时,发现其为双链,分子量约为1.1×10⁶。这些颗粒在4℃的CsCl中至少5个月保持稳定。电子显微镜照片显示,用2%磷钨酸染色的高度纯化的PLP与多瘤病毒衣壳外观相同。由松散离子键结合形成的聚集体或复合物似乎都无法合理解释这些结果。证据表明,在无细胞条件下形成的这种新的多瘤样颗粒的DNA受到衣壳的保护。