Quadt R, Rosdorff H J, Hunt T W, Jaspars E M
Department of Biochemistry, Gorlaeus Laboratories, Leiden University, The Netherlands.
Virology. 1991 May;182(1):309-15. doi: 10.1016/0042-6822(91)90674-z.
RNA-dependent RNA polymerase (RdRp) was solubilized and purified from cellular membranes isolated from alfalfa mosaic virus (AIMV)-infected tobacco by employing a procedure recently described for brome mosaic virus RdRp [R. Quadt and E.M.J. Jaspars, 1990, Virology 178, 189-194]. The purified AIMV RdRp is completely dependent on added template RNAs and exhibits a high degree of template specificity. Analysis of the protein composition of AIMV RdRp showed that AIMV-encoded proteins P1 and P2 and the coat protein (CP) are present in the active enzyme complex. Minus-strand synthesis by the AIMV RdRp is inhibited by AIMV CP. Native double-stranded AIMV RNAs are utilized as template for viral RNA synthesis by AIMV RdRp indicating that a helicase activity is present in the purified AIMV RdRp preparation.
通过采用最近描述的用于雀麦花叶病毒RdRp的方法[R. 夸特和E.M.J. 贾斯帕斯，1990年，《病毒学》178卷，第189 - 194页]，从感染苜蓿花叶病毒（AIMV）的烟草中分离的细胞膜中溶解并纯化了RNA依赖性RNA聚合酶（RdRp）。纯化的AIMV RdRp完全依赖于添加的模板RNA，并表现出高度的模板特异性。对AIMV RdRp蛋白质组成的分析表明，AIMV编码的蛋白质P1和P2以及衣壳蛋白（CP）存在于活性酶复合物中。AIMV RdRp的负链合成受到AIMV CP的抑制。天然双链AIMV RNA被用作AIMV RdRp进行病毒RNA合成的模板，这表明在纯化的AIMV RdRp制剂中存在解旋酶活性。
