title: Expression of L protein of Hantaan virus 84FLi strain and its application for recovery of minigenomes.
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.
authors.affiliation: Center of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an, P. R. China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xin-Hong
authors.initials: XH
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Jiang
authors.forename: Chang-Xing
authors.initials: CX
authors.lastname: Huang
authors.forename: Ping-Zhong
authors.initials: PZ
authors.lastname: Wang
authors.forename: Dan-Lei
authors.initials: DL
authors.lastname: Mou
authors.forename: Li
authors.initials: L
authors.lastname: Sun
authors.forename: Zhe
authors.initials: Z
authors.lastname: Xu
authors.forename: Xin
authors.initials: X
authors.lastname: Wei
authors.forename: Xue-Fan
authors.initials: XF
authors.lastname: Bai
mesh_terms: D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis; D018063:Hantaan virus / Q000235:genetics / Q000502:physiology*; D018778:Hantavirus Infections / Q000821:virology*; D006801:Humans; D012318:RNA Polymerase I / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000032:analysis* / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D048348:Reverse Transcription; D014709:Vero Cells / Q000378:metabolism; D014764:Viral Proteins / Q000032:analysis* / Q000096:biosynthesis
chemical_list: D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D015500:Chloramphenicol O-Acetyltransferase; C500570:L protein, hantavirus; D012324:RNA-Dependent RNA Polymerase; D012318:RNA Polymerase I