BIOCHEMICAL-CHARACTERIZATION OF PORCINE ENTERIC CALICIVIRUS - ANALYSIS OF STRUCTURAL AND NONSTRUCTURAL VIRAL-PROTEINS
BIOCHEMICAL-CHARACTERIZATION OF PORCINE ENTERIC CALICIVIRUS - ANALYSIS OF STRUCTURAL AND NONSTRUCTURAL VIRAL-PROTEINS
复制标题
DOI：
10.1007/bf01348984
复制
发表时间：
1990-01-01
影响因子：
2.7
通讯作者：
KANG, SY
中科院分区：
文献类型：
--
作者：
PARWANI, AV;SAIF, LJ;KANG, SY
本文对猪肠道杯状病毒（PEC）的蛋白质分子量和抗原性进行了研究。PEC病毒体从感染猪的肠内容物和感染的细胞培养裂解物中纯化。纯化病毒在氯化铯中的平均浮力密度为1.37 gm/cm 3。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PEC中发现一种分子量约为58 K的主要结构蛋白。使用免疫印迹技术，只有一个免疫反应蛋白（58 k）被确定。PEC和原型杯状病毒猫杯状病毒（FCV）分别在猪肾和猫肾（Crandell）细胞系中繁殖，并在接种后（PI）的不同时间使用[35 S]甲硫氨酸进行固有标记。放射性标记的蛋白质的SDS-PAGE表明，存在的主要结构蛋白（58 k）和一个可能的非结构蛋白（28 k）的PEC感染的细胞裂解物中合成的PI 12小时。其他次要蛋白条带在24 h PI（32 k和82 k）时也很明显。只有58 k主要蛋白检测放射免疫沉淀（RIP）分析使用超免疫抗PEC血清。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，鉴定出约64 k. RIP分析表明PEC和FCV蛋白之间没有抗原关系。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1.37 gm/cm 3的平均密度与杯状病毒科其他成员的特性一致。
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) was 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.