Parwani A V, Saif L J, Kang S Y
Ohio Agricultural Research and Development Center, Ohio State University, Wooster.
Arch Virol. 1990;112(1-2):41-53. doi: 10.1007/BF01348984.
In this report, the molecular weight and antigenicity of the proteins of a porcine enteric calicivirus (PEC) were characterized. The PEC virions were purified from intestinal contents of infected pigs and from infected cell culture lysates. The average buoyant density of the purified virus was 1.37 gm/cm3 in cesium chloride. One major structural protein with a molecular weight of approximately 58 k was found in the gut and cell culture-passaged PEC using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Using immunoblotting techniques only one immunoreactive protein (58 k) ws identified. The PEC and a prototype calicivirus, feline calicivirus (FCV) were propagated in pig kidney and feline kidney (Crandell) cell lines, respectively and intrinsically labeled using [35S]methionine at various times post-inoculation (PI). SDS-PAGE of the radiolabeled proteins indicated the presence of the major structural protein (58 k) and one probable nonstructural protein (28 k) synthesized in the PEC-infected cell lysates by 12 h PI. Other minor protein bands were also evident by 24 h PI (32 k and 82 k). Only the 58 k major protein was detected by radioimmunoprecipitation (RIP) analysis using hyperimmune anti-PEC serum. SDS-PAGE and RIP analysis of FCV-infected cell lysates using hyperimmune anti-FCV serum identified a single major protein of approximately 64 k. No antigenic relationship between PEC and FCV proteins was detected by RIP analysis. The single major structural protein of PEC, the morphological appearance and size of the virus, and its average density of 1.37 gm/cm3 in cesium chloride are consistent with properties of other members of the family Caliciviridae.
在本报告中，对一种猪肠道杯状病毒（PEC）的蛋白质的分子量和抗原性进行了表征。PEC病毒粒子从感染猪的肠道内容物和感染的细胞培养裂解物中纯化得到。在氯化铯中，纯化病毒的平均浮力密度为1.37 g/cm³。使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）在肠道和细胞培养传代的PEC中发现了一种分子量约为58 kD的主要结构蛋白。使用免疫印迹技术仅鉴定出一种免疫反应性蛋白（58 kD）。PEC和一种杯状病毒原型——猫杯状病毒（FCV）分别在猪肾和猫肾（克兰德尔）细胞系中繁殖，并在接种后不同时间点使用[³⁵S]甲硫氨酸进行体内标记。放射性标记蛋白的SDS-PAGE表明，在接种后12小时，PEC感染的细胞裂解物中合成了主要结构蛋白（58 kD）和一种可能的非结构蛋白（28 kD）。到接种后24小时，其他次要蛋白条带也很明显（32 kD和82 kD）。使用超免疫抗PEC血清通过放射免疫沉淀（RIP）分析仅检测到58 kD的主要蛋白。使用超免疫抗FCV血清对FCV感染的细胞裂解物进行SDS-PAGE和RIP分析，鉴定出一种约64 kD的单一主要蛋白。通过RIP分析未检测到PEC和FCV蛋白之间的抗原关系。PEC的单一主要结构蛋白、病毒的形态外观和大小以及其在氯化铯中的平均密度1.37 g/cm³与杯状病毒科其他成员的特性一致。
