Cloning of a gene encoding Lymantria xylina nucleopolyhedrovirus fusion protein and its expression in LD cells
Date Issued
2005
Date
2005
Author(s)
Pien, Hsiu-Wen
DOI
zh-TW
Abstract
IPLB LD-652Y-5d cells infected with Lymantria xylina multiple nucleopolyhedrovirus (LyxyMNPV) were found that the infection cells were fused together and formed syncytial giant cells. This phenomenon is probably caused by the viral envelope fusion protein in the cell membrane of the infected cells. Recently, two viral genes encoding the envelope fusion proteins, GP64 and LD130, had been identified. Both GP64 and LD130 envelope fusion proteins mediate the cell fusion at low pH value. The envelope fusion protein of NPV Group I is GP64 while Group II is LD130. We cloned and sequenced ld130 from LyxyMNPV. The LyxyNPV ld130 consists of 2,025 bp. Comparison of the nucleotide and amino acid sequences of LyxyMNPV ld130 with those of LdMNPV ld130 showed 93 and 96% identities, respectively. The predicted peptide sequence of LyxyMNPV LD130 contains signal and transmembrane domains that are significant homology to membrane proteins. Analysis of LyxyNPV ld130 transcript revealed both early and late transcriptional initiation sites located at upstream of the 5’ ATG initiation codon. In addition, a poly A sequence located at 17 nt downstream of the 3’ polyadenylation signal (ATTAAA). Based on the sequences of ld130 gene, the NPV species can be divided into two Groups, I and II, in phylogenetic analysis, LyxyMNPV belongs to Group II. The LD130 of budded virus detected by Western blot revealed that LD130 is a viral structural protein. The LD130 were constructed with the promoter of immediately early gene and expressed by baculovirus transient expression vector in uninfected insect cells showed that LD130 could mediate the cell-to-cell fusion at low pH. These results suggest that a functional homolog of LD130 envelope fusion protein in group II NPVs was found in LyxyMNPV.
Subjects
黑角舞蛾核多角體病毒
套膜融合蛋白
細胞融合
Lymantria xylina nucleopolyhedrovirus
envelope fusion protein
ld130
Type
thesis
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