IDENTIFICATION OF A 17000 MOLECULAR-WEIGHT ANTIGENIC POLYPEPTIDE IN TRANSMISSIBLE GASTROENTERITIS VIRUS-INFECTED CELLS
IDENTIFICATION OF A 17000 MOLECULAR-WEIGHT ANTIGENIC POLYPEPTIDE IN TRANSMISSIBLE GASTROENTERITIS VIRUS-INFECTED CELLS
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DOI：
10.1099/0022-1317-67-7-1419
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发表时间：
1986-07-01
影响因子：
3.8
通讯作者：
WOODS, RD
中科院分区：
文献类型：
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作者：
WESLEY, RD;WOODS, RD
在感染后不同时间用[35S]蛋氨酸对细胞进行脉冲标记，然后用SDS-PAGE分离和鉴定传染性胃肠炎病毒（TGEV）感染猪睾丸（ST）细胞培养物特异性多肽。感染后6 h，在放射性标记细胞提取物中检测到TGEV的主要结构蛋白，表观分子量分别为200000 （200K）、47K和30K。此外，在感染细胞中存在17K主要多肽，但在模拟感染的对照培养中不存在。用[3H]氨基葡萄糖标记显示只有200K和30K蛋白被糖基化。在体外用蔗糖梯度纯化病毒二次刺激tgevv引物的猪淋巴细胞，只产生针对两种糖蛋白（gp）的抗体，这表明17K多肽不是病毒粒子的表面特征。对2头猪经口感染TGEV米勒毒株，采用免疫沉淀法对其血清进行分析。在第25天，港口感染恢复期血清对gp30和gp200反应强烈，对17K多肽的初始反应较弱。感染后60天血清免疫球蛋白对17K蛋白反应强烈，而对gp30的抗体反应明显降低，对gp200的抗体反应略有降低。
PUlse labelling of cells with [35S]methionine at different times after infection followed by SDS-PAGE was used to resolve and to identify polypeptides designated as specific to transmissible gastroenteritis virus (TGEV)-infected swine testicular (ST) cell cultures. The major TGEV structural proteins, with apparent molecular weights of 200000 (200K), 47K and 30K were detected in radiolabelled cell extracts by 6 h post-infection. Additionally, a 17K major polypeptide was present in infected cells but not in mock-infected control cultures. Labelling with [3H]glucosamine revealed only the 200K and 30K proteins to be glcosylated. TGEV-primed porcine lymphocytes, secondarily stimulated in vitro with sucrose gradient-purified virus, produced antibody only to the two glycoproteins (gp) indicating that the 17K polypeptide is not a surface feature of the virion. Two pigs were infected oronasally with the virulent Miller strain of TGEV and their sera were analysed by immunoprecipitation. At 25 days port-infection convalescent sera responded strongly to gp30 and gp200 and there was a weak initial response to the 17K polypeptide. Serum immunoglobulins at 60 days post-infection reacted stronly to the 17K protein while the antibody response to gp30 was significantly reduced and that to gp200 was slightly reduced.