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多瘤病毒衣壳蛋白VP1和VP2的无细胞合成

Cell-free synthesis of polyoma virus capsid proteins VP1 and VP2.

作者信息

Wheeler T, Bayley S T, Harvey R, Crawford L V, Smith A E

出版信息

J Virol. 1977 Jan;21(1):215-24. doi: 10.1128/JVI.21.1.215-224.1977.

Abstract

Polyadenylated RNA isolated from the cytoplasm of mouse 3T6 cells 28 h after infection with polyoma virus has been isolated and translated in vitro. Polyoma capsid proteins VP1 and VP2 have been identified in the cell-free product by polyacrylamide gel electrophoresis, specific immunoprecipitation, and tryptic peptide fingerprinting. Polyoma mRNA species have been isolated by preparative hybridization to purified viral DNA immobilized on cellulose nitrate filters and shown to code for both VP1 and VP2. These experiments establish conditions for the isolation of late polyoma mRNA and the cell-free synthesis of polyoma capsid proteins and indicate that the active mRNA species are at least partially virus coded.

摘要

从多瘤病毒感染小鼠3T6细胞28小时后的细胞质中分离出的多聚腺苷酸化RNA,已被分离并在体外进行翻译。通过聚丙烯酰胺凝胶电泳、特异性免疫沉淀和胰蛋白酶肽指纹图谱分析,在无细胞产物中鉴定出多瘤病毒衣壳蛋白VP1和VP2。通过与固定在硝酸纤维素滤膜上的纯化病毒DNA进行制备性杂交,分离出多瘤病毒mRNA种类,并证明其编码VP1和VP2。这些实验建立了分离晚期多瘤病毒mRNA和无细胞合成多瘤病毒衣壳蛋白的条件,并表明活性mRNA种类至少部分由病毒编码。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6021/353807/2aeeaf536656/jvirol00205-0232-a.jpg

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