STRUCTURE AND CHEMICAL-PHYSICAL CHARACTERISTICS OF LACTATE DEHYDROGENASE-ELEVATING VIRUS AND ITS RNA
STRUCTURE AND CHEMICAL-PHYSICAL CHARACTERISTICS OF LACTATE DEHYDROGENASE-ELEVATING VIRUS AND ITS RNA
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DOI：
10.1128/jvi.16.2.420-433.1975
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发表时间：
1975-01-01
影响因子：
5.4
通讯作者：
PLAGEMANN, PGW
中科院分区：
文献类型：
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作者：
BRINTONDARNELL, M;PLAGEMANN, PGW
乳酸脱氢酶升高病毒（LDV）是从小鼠各种组织（腹膜巨噬细胞、骨髓、脾脏和胚胎）感染原代培养物的培养液和感染小鼠的血浆中纯化出来的。阴性染色的病毒和阳性染色的LDV切片电镜显示，病毒呈直径约55 nm的均匀球形颗粒，含有直径约30 nm的电子致密核。在样品制备过程中，包膜有脱落和解体的趋势，形成各种大小的聚集体和直径为8至14纳米的小空心颗粒。两株LDV在不同小鼠组织的原代培养物中繁殖或从感染小鼠的血浆中分离得到的密度均为1.13 g/cm3。LDV在含有0.01% Nonidet P-40或Triton X的溶液中短暂孵育足以释放病毒的核衣壳，而类似的处理对Sindbis病毒没有影响。LDV的核壳密度为1.17 g/cm3，不含磷脂酰胆碱，只含有最小的病毒蛋白VP-1，分子量约为15000。包膜含有两种蛋白质。VP-2分子量为18000，糖蛋白VP-3在聚丙烯酰胺凝胶电泳过程中异质性迁移（24000 - 44000道尔顿）。与29S rRNA的沉降速率相比，LDV和Sindbis病毒的rna分别在48和45S沉降，无论是在低盐浓度或高盐浓度的蔗糖密度梯度下进行带沉降分析，还是在甲醛处理下进行变性。我们的结果表明，LDV应该被归类为托加病毒，但LDV与α病毒和黄病毒有很大的不同，因此被排除在这些组之外。
Lactate dehydrogenase-elevating virus (LDV) was purified from culture fluid of infected primary cultures of various mouse tissues (peritoneal macrophage, bone marrow, spleen, and embryo) and from plasma of infected mice. Electron microscopy of negatively stained virus and positively stained sections of LDV revealed spherical particles of uniform size with a diameter of about 55 nm, containing an electron-dense core with a diameter of about 30 nm. During sample preparation the envelope had a tendency to slough off and disintegrate to form aggregates of various sizes and small hollow particles with a diameter of 8 to 14 nm. Two strains of LDV exhibited a density of 1.13 g/cm3 in isopycnic sucrose density gradient centrifugation whether propagated in primary cultures of the various mouse tissues or isolated from plasma of infected mice. A brief incubation of LDV in a solution containing 0.01% Nonidet P-40 or Triton X was sufficient to release the viral nucleocapsid, whereas a similar treatment had no effect on Sindbis virus. The nucleocapdis of LDV exhibited a density of 1.17 g/cm3, was devoid of phosphatidylcholine, and contained only the smallest of the viral proteins, VP-1, which had a molecular weight of about 15,000. The envelope contained two proteins. VP-2 with a molecular weight of 18,000 and a glycoprotein, VP-3, which migrated heterogenously (24,000 to 44,000 daltons) during polyacrylamide gel electrophoresis. When compared to the sedimentation rate of 29S rRNA, the RNAs of LDV and Sindbis virus sedimented at 48 and 45S, respectively, whether analyzed by zone sedimentation in sucrose density gradients containing low or high salt concentrations or denatured by treatment with formaldehyde. Our results indicate that LDV should be classified as a togavirus, but that LDV is sufficiently different from alpha and flaviviruses to be excluded from these groups.