Journal of Virology (1998) 72, 10093-10099

Yi-Ija Li, Yueh-Mei Cheng, Yih-Leh Huang, Ching-Hsiu Tsai, Yau-Heiu Hsu and Menghsiao Meng (1998)
Identification and characterization of the Escherichia coli-expressed RNA-dependent RNA polymerase of Bamboo mosaic virus
Journal of Virology 72 (12), 10093-10099
Abstract: Bamboo mosaic virus (BaMV), a member of the potexvirus group, infects primarily members of the Bambusoideae. The open reading frame 1 (ORF1) of BaMV encodes a 155-kDa polypeptide that was postulated to be involved in the replication and the formation of cap structure at the 5' end of the viral genome. To characterize the activities associated with the 155-kDa viral protein, it was expressed in Escherichia coli BL21(DE3) cells with thioredoxin, hexahistidine, and S · Tag fused consecutively at its amino terminus, and the fusion protein was purified by metal affinity chromatography. Several RNA fragments, prepared by in vitro transcription, were tested as substrates for the RNA-dependent RNA polymerase (RdRp) activity. Among them, the expressed fusion enzyme was able to generate a ³²P-labeled RNA product when 3'-end RNA fragments of the positive strand or negative strand of BaMV were included in the assay mixture. Dot hybridization assay revealed that the reaction products are complementary to their RNA substrates. Taken together, the evidence suggests that the 155-kDa protein encoded by ORF1 of BaMV has an RdRp activity and should be involved in the replication of BaMV. Mutational analyses demonstrate the importance of the GDD motif in the polymerase activity, and deletion studies suggest that the polymerase activity resides in the carboxyl terminus of the 155-kDa viral protein.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Full text of article
Database assignments for author(s): Ching-Hsiu Tsai, Menghsiao Meng

Research topic(s) for pests/diseases/weeds:
molecular biology - genes

---

Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Bamboo mosaic virus