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麻疹病毒基质基因的克隆与体外表达

Cloning and in vitro expression of the measles virus matrix gene.

作者信息

Greer P A, Hasel K W, Millward S

出版信息

Biochem Cell Biol. 1986 Oct;64(10):1038-43. doi: 10.1139/o86-137.

Abstract

A cDNA library was prepared from Vero cells infected with the Edmonston strain of measles virus. A number of viral specific cDNA clones were isolated and characterized by Northern blot hybridization analysis. A cDNA clone containing a 1500 base pair insert which hybridizes to a viral specific transcript of approximately 1500 nucleotides was subcloned into pSP64 and used as an in vitro transcription template. The resulting RNA transcript was translated in a cell-free system, giving rise to a polypeptide which comigrates on polyacrylamide gels with the authentic measles virus matrix protein and is immunoprecipitated with antisera specific for the matrix protein.

摘要

从感染麻疹病毒埃德蒙斯顿株的非洲绿猴肾细胞(Vero细胞)制备了一个互补DNA(cDNA)文库。通过Northern印迹杂交分析分离并鉴定了许多病毒特异性cDNA克隆。将一个含有1500个碱基对插入片段的cDNA克隆亚克隆到pSP64中,该插入片段与一个约1500个核苷酸的病毒特异性转录本杂交,并用作体外转录模板。所得的RNA转录本在无细胞系统中进行翻译,产生一种多肽,该多肽在聚丙烯酰胺凝胶上与真正的麻疹病毒基质蛋白迁移率相同,并且能用针对基质蛋白的抗血清进行免疫沉淀。

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