Salmeen I, Rimai L, Luftig R B, Libes L, Retzel E, Rich M, McCormick J J
J Virol. 1976 Feb;17(2):584-96. doi: 10.1128/JVI.17.2.584-596.1976.
We have studied purified preparations of murine mammary tumor virus (MuMTV), Rous sarcoma virus (RSV; Prague strain), and feline leukemia virus (FeLV) by laser beat frequency light-scattering spectroscopy, ultra-centrifugation, and electron microscopy. The laser beat frequency light-scattering spectroscopy measurements yield the light-scattering intensity, weighted diffusion coefficients. The corresponding average hydrodynamic diameters, as calculated from the diffusion coefficients by the Stokes-Einstein equation for MuMTV, RSV, and FeLV, respectively, are: 144 +/- 6 nm, 147 +/- 7 nm, and 168 +/- 6 nm. Portions of the purified RSV and MuMTV preparations, from which light-scattering samples were obtained, and portions of the actual FeLV light-scattering samples were examined by negatively stained, catalase crystal-calibrated electron microscopy. The light-scattering intensity weighted averages of the electron micrograph size distributions were calculated by weighing each size by its theoretical relative scattering intensity, as obtained from published tables computed according to the Mie scattering theory. These averages and the experimentally observed hydrodynamic diameters agreed to within +/- 5%, which is the combined experimental error in the electron microscopic and light-scattering techniques. We conclude that the size distributions of singlet particles observed in the electron micrographs are statistically true representations of the sedimentation-purified solution size distributions. The sedimentation coefficients (S20, w) for MuMTV, RSV, and FeLV, respectively, are: 595 +/- 29S, 689 +/- 35S, and 880 +/- 44S. Virus partial specific volumes were taken as the reciprocals of the buoyant densities, determined in sucrose density gradients. The Svedberg equation was used to calculate particle weights from the measured diffusion and sedimentation coefficients. The particle weights for MuMTV, RSV, and FeLV, respectively, are: (3.17 +/- 0.32) x 10(8), (4.17 +/- 0.42) x 10(8), and (5.50 +/- 0.55) x 10(8) daltons.
我们通过激光拍频光散射光谱法、超速离心法和电子显微镜,对纯化的小鼠乳腺肿瘤病毒(MuMTV)、劳氏肉瘤病毒(RSV;布拉格株)和猫白血病病毒(FeLV)制剂进行了研究。激光拍频光散射光谱测量得出了光散射强度、加权扩散系数。根据斯托克斯 - 爱因斯坦方程分别由MuMTV、RSV和FeLV的扩散系数计算出的相应平均流体动力学直径为:144±6纳米、147±7纳米和168±6纳米。对用于获取光散射样品的纯化RSV和MuMTV制剂部分,以及实际的FeLV光散射样品部分进行了经负染色、过氧化氢酶晶体校准的电子显微镜检查。通过根据米氏散射理论计算的已发表表格得出的理论相对散射强度对每个尺寸进行加权,计算出电子显微镜图像尺寸分布的光散射强度加权平均值。这些平均值与实验观察到的流体动力学直径的误差在±5%以内,这是电子显微镜和光散射技术的综合实验误差。我们得出结论,在电子显微镜图像中观察到的单颗粒尺寸分布在统计学上真实反映了沉降纯化溶液的尺寸分布。MuMTV、RSV和FeLV的沉降系数(S20,w)分别为:595±29S, 689±35S, 和880±44S。病毒的偏比容取蔗糖密度梯度中测定的浮力密度的倒数。使用斯维德贝格方程根据测得的扩散系数和沉降系数计算颗粒重量。MuMTV、RSV和FeLV的颗粒重量分别为:(3.17±0.32)×10⁸、(4.17±0.42)×10⁸和(5.50±0.55)×10⁸道尔顿。