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编码结构蛋白的风疹病毒互补脱氧核糖核酸的表达

Expression of rubella virus cDNA coding for the structural proteins.

作者信息

Clarke D M, Loo T W, McDonald H, Gillam S

机构信息

Department of Pathology, Faculty of Medicine, University of British Columbia, Vancouver, Canada.

出版信息

Gene. 1988 May 15;65(1):23-30. doi: 10.1016/0378-1119(88)90413-1.

Abstract

A cDNA clone encoding the precursor polypeptide (Mr 115,000) to the nucleocapsid C (Mr 30,000) and two envelope glycoproteins E1 (Mr 58,000) and E2 (Mr 42,000-47,000) of rubella virus was inserted into a simian virus 40-derived eukaryotic expression vector. When the plasmid was introduced into COS cells, three proteins were synthesized. The expressed proteins were antigenically similar and identical in size to the authentic structural proteins of rubella virus. Expression in the presence of tunicamycin confirmed that E1 and E2 are glycoproteins. Unglycosylated E1 and E2 had Mrs of about 53,000 and 30,000, respectively. The mobility of the nucleocapsid protein was unaffected by tunicamycin. The locations of the translation start and stop codons for synthesis of the precursor to the structural proteins of rubella virus were determined by in vitro and in vivo expression studies. It was found that the first AUG codon at the 5' end of the rubella virus 24S cDNA acts as a start codon for translation. The stop codon was found to be 3183 bp from the start codon.

摘要

将编码风疹病毒核衣壳蛋白C(分子量30,000)的前体多肽(分子量115,000)以及两种包膜糖蛋白E1(分子量58,000)和E2(分子量42,000 - 47,000)的cDNA克隆插入到猿猴病毒40衍生的真核表达载体中。当将该质粒导入COS细胞时,合成了三种蛋白质。所表达的蛋白质在抗原性上与风疹病毒的天然结构蛋白相似且大小相同。在衣霉素存在下的表达证实E1和E2是糖蛋白。未糖基化的E1和E2的分子量分别约为53,000和30,000。核衣壳蛋白的迁移率不受衣霉素影响。通过体外和体内表达研究确定了风疹病毒结构蛋白前体合成的翻译起始和终止密码子的位置。发现风疹病毒24S cDNA 5'端的第一个AUG密码子作为翻译的起始密码子。发现终止密码子距起始密码子3183 bp。

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