Akademik Veri Yönetim Sistemi
JOURNAL OF APPLIED BIOLOGICAL SCIENCES, cilt.18, sa.3, ss.265-276, 2024 (Hakemli Dergi)
The Amsacta moorei entomopoxvirus (AMEV) belongs to the Entomopoxvirinae
subfamily of poxviruses. Although entomopoxviruses are very similar to
poxviruses of humans, they have no potential risk for humans because
they only infect insects. Intensive studies have been carried out on
this virus because it can potentially be used as gene therapy and gene
expression vectors. The AMEV genome has 256 open reading frames and two
of them, AMV153 and AMV197, are putative protein
kinases. Protein kinases of some vertebrate poxviruses have some
functions in virus morphogenesis, regulation of the cell cycle, and
apoptosis of the host. In previous studies, the molecular structure of AMV197
protein kinase was elucidated, its transcriptomic properties in cell
culture and its function in progeny virus production were determined,
and an AMV197-defective virus (AmΔPK/gfp) was produced. However, the function of AMV153
in this scenario remained as unclear, and it is crucial to know its
function to better understand the role of protein kinases in the
replication of AMEV. The AMV153 was first deleted from the genome of AmΔPK/gfp
and subsequently from the genome of AMEV, creating two separate
recombinant viruses. The homologous recombination method was used to
replace AMV153 ORF with mCherry gene that produces red fluorescent protein. Mutant viruses in which the AMV153 gene was replaced with mCherry were identified by fluorescence microscopy but could not be propagated separately from the AmΔPK/gfp or wild-type viruses in insect cells. Unsuccessful attempts to isolate the mutant viruses with the AMV153 gene deletion first in AmΔPK/gfp virus and second in wild type AMEV structure, suggested that the protein kinase encoded by AMV197 cannot substitute the function of AMV153 and the AMV153 protein is essential for virus replication.