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자료유형
학술저널
저자정보
저널정보
대한생화학·분자생물학회 Experimental and Molecular Medicine Experimental and Molecular Medicine 제35권 제6호
발행연도
2003.1
수록면
475 - 485 (11page)

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The hepatitis C virus (HCV) RNA-dependent RNA polymerase, NS5B protein, is the key viral enzyme responsible for replication of the HCV viral RNA genome. Although several ful-length and trun-cated forms of the HCV NS5B proteins have been expressed previously in insect cels, contamina-tion of host terminal transferase (TNTase) has hampered analysis of the RNA synthesis initiation mechanism using natural HCV RNA templates. We have expressed the HCV NS5B protein in insect it to near homogeneity without contaminated TNT-ase. The highly purified recombinant HCV NS5B was capable of copying 9.6-kb ful-length HCV RNA template, and mini-HCV RNA carying both 5'- and 3'-untranslated regions (UTRs) of the HCV ge-nome. In the absence of a primer, and other cellul-ar and viral factors, the NS5B could elongate over HCV RNA templates, but the synthesized products were primarily in the double stranded form, indica-ting that no cyclic replication occured with NS5B alone. RNA synthesis using RNA templates repre-RNA and the X-RNA at the 3'-end of HCV RNA genome was also initiated de novo. No formation of dimer-size self-primed RNA products resulting from extension of the 3'-end hydroxyl group was observed. Despite the internal de novo initiation from the X-RNA, the NS5B could not initiate RNA synthesis from the internal region of oligouridylic acid (U)20, sugesting that HCV RNA polymerase initiates RNA synthesis from the selected region in the 3'-UTR of HCV genome.

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