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Expression of L protein of Hantaan virus 84FLi strain and its application for recovery of minigenomes

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Abstract:

Zhang Y, Li XH, Jiang H, Huang CX, Wang PZ, Mou DL, Sun L, Xu Z, Wei X, Bai XF. Expression of L protein of Hantaan virus 84FLi strain and its application for recovery of minigenomes. APMIS 2008;116:1089–96.

Hantaan virus (HTNV) is an Old World hantavirus associated with hemorrhagic fever with renal syndrome (HFRS). To visualize the localization of the L protein of HTNV strain 84FLi within cells, a fusion protein composed of enhanced green fluorescent protein and L protein, EGFP-L, was expressed in Vero cells. The 273 KDa expressed fusion protein of EGFP-L localized in the perinuclear region. We also described the development of a reverse genetics system for HTNV strain 84FLi. The RNA polymerase I (pol I)-mediated transcription system was used to generate artificial viral RNA genome segments (minigenomes), which contained the chloramphenicol acetyltransferase (CAT) reporter gene in antisense (virus RNA) or sense (virus-complementary RNA) orientation flanked by the noncoding regions of HTNV 84FLi L segment. CAT could be detected in cells after transfection, indicating the successful encapsidation, transcription and replication of the pol I-derived minigenomes. The passaged transfer of CAT demonstrates that recombinant virus containing packaged pol I-derived minigenomes has been produced. This system may be helpful in studying the gene function and pathogenesis of HTNV.

Keywords: Hantaan virus; RNA polymerase I; RNA-dependent RNA polymerase; minigenomes; reverse genetics

Document Type: Original Article

DOI: http://dx.doi.org/10.1111/j.1600-0463.2008.01011.x

Publication date: December 1, 2008

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