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- Title
Expression of L protein of Hantaan virus 84FLi strain and its application for recovery of minigenomes.
- Authors
YE ZHANG; XIN-HONG LI; HONG JIANG; CHANG-XING HUANG; PING-ZHONG WANG; DAN-LEI MOU; LI SUN; ZHE XU; XIN WEI; XUE-FAN BAI
- Abstract
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.
- Subjects
HANTAVIRUS diseases; HEMORRHAGIC fever; KIDNEY diseases; GREEN fluorescent protein; ANTISENSE RNA
- Publication
APMIS, 2008, Vol 116, Issue 12, p1089
- ISSN
0903-4641
- Publication type
Article
- DOI
10.1111/j.1600-0463.2008.01011.x