2019 ; Vol.29-11: 1790~1798
|Author||Doyeong Kim, Tae-Ju Cho|
|Place of duty||School of Life Sciences, Chungbuk National University, Cheongju 28644, Republic of Korea|
|Title||Flock House Virus RNA1 with a Long Heterologous Sequence at the 3’-end Can Replicate in Mammalian Cells and Mediate Reporter Gene Expression|
J. Microbiol. Biotechnol.2019 ;
|Abstract||Flock House virus (FHV), an insect RNA virus, has a bipartite genome. FHV RNA1 can be
packaged in turnip yellow mosaic virus (TYMV) as long as the FHV RNA has a TYMV
sequence at the 3’-end. The encapsidated FHV RNA1 has four additional nucleotides at the 5’-
end. We investigated whether the recombinant FHV RNA1 could replicate in mammalian
cells. To address this issue, we prepared in vitro transcribed FHV RNAs that mimicked the
recombinant FHV RNA1, and introduced them into baby hamster kidney (BHK) cells. The
result showed that the recombinant FHV RNA1 was capable of replication. An eGFP gene
inserted into the frame with B2 gene of the FHV RNA1 was also successfully expressed. We
also observed that eGFP expression at the protein level was strong at 28oC but weak at 30oC.
Sequence analysis showed that the 3’-ends of the RNA1 and RNA3 replication products were
identical to those of the authentic FHV RNAs. This indicates that FHV replicase correctly
recognized an internally-located replication signal. In contrast, the 5’-ends of recombinant
FHV RNA1 frequently had deletions, indicating random initiation of (+)-strand synthesis.|
|Key_word||Turnip yellow mosaic virus, Flock house virus, FHV RNA1, FHV RNA replication, reporter gene expression|
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