Jensen M J, Moore D M
Molecular Biology Laboratory, Plum Island Animal Disease Center, USDA-Agricultural Research Service, Greenport, New York 11944.
Virology. 1993 Apr;193(2):604-13. doi: 10.1006/viro.1993.1168.
A series of genetically related variants arising from a parental wild-type isolate of O1 Campos and its tissue culture adapted variant were differentiated by various cell culture markers (temperature sensitivity, plaque size, viral yield) and lethality in mice. These isolates were additionally characterized functionally and biochemically by examining poly(C) length, RNA synthesis, protein synthesis, and cell receptor binding. In primary bovine kidney cells, the virulent isolates had greater levels of protein synthesis, whereas in baby hamster kidney cells, the attenuated variant outproduced the wild-type parent. The tissue culture adapted variant had substantially greater ability to attach to cells than the parental wild type. The parental wild-type and the tissue culture-adapted variant were similarly neutralized by various sera against whole virus, but the parental wild type was less effectively neutralized by sera prepared from either full or truncated (variable region) bacterially expressed VP1 polypeptides. The capsid region of the genomes of both these variants was sequenced and a nucleotide substitution resulting in a change in amino acid 56 in VP3 was found. The nucleotide sequence change for the remaining two variants was that of the parental wild-type virus.
一系列源自O1坎波斯亲本野生型分离株及其适应组织培养的变体，通过各种细胞培养标记（温度敏感性、蚀斑大小、病毒产量）以及对小鼠的致死性进行区分。通过检测聚（C）长度、RNA合成、蛋白质合成和细胞受体结合，对这些分离株进行了功能和生化特性的进一步表征。在原代牛肾细胞中，强毒株的蛋白质合成水平更高，而在幼仓鼠肾细胞中，减毒株的产量超过野生型亲本。适应组织培养的变体比亲本野生型具有更强的细胞附着能力。亲本野生型和适应组织培养的变体被各种抗全病毒血清同样程度地中和，但亲本野生型被由全长或截短（可变区）细菌表达的VP1多肽制备的血清中和的效果较差。对这两种变体的基因组衣壳区域进行了测序，发现一个核苷酸替换导致VP3中第56位氨基酸发生变化。其余两种变体的核苷酸序列变化与亲本野生型病毒相同。
