title: Molecular and immunohistochemical characterization of granulin gene encoded in Pieris rapae granulovirus genome.
abstract: Pieris rapae granulovirus (PiraGV) is highly pathogenic to the cabbage butterfly (P. rapae), an important pest of cultivated cabbages and mustard crops. It therefore holds significant promise towards exploitation as a potent bio-control agent in the field controlling the pest population. Whole-genome elucidation of the Korean isolate of the granulovirus (PiraGV-K), reported the presence of a granulin gene corresponding to ORF 1 in its genome. Comprehensive studies towards functional characterization of the gene, established that it is composed of 744 nucleotides and encodes a peptide of 247 amino acid residues. It possessed significant homology with AoGV and ClanGV with 87% identity at amino acid level. Multiple alignment data suggests that the C-terminus region of the gene had three different conserved regions. Time-course studies conducted in PiraGV-K infected P. rapae larvae revealed a significant upsurge of the transcript (134-fold) at 4 days post infection followed by a significant decline at the most advanced stages of infection. Anti-PiraGV-K granulin antibody was produced and western blot conducted with the infected larvae further confirmed the induction pattern with a protein of 30 kDa. Immunofluorescent staining showed a granulin-specific signal in fat body and integument of the infected larvae. Granulin-specific signals were noticed 2 days post infection with the eventual systemic spread of infection to the associated tracheal matrix witnessed at 4 days post infection. Immunogold labeling and electron microscopic studies further proved the cytopathological effects as the presence of numerous membrane-bound vesicles with nucleocapsids and abruption of intercellular junctions in fat body and hypertrophied cells in the integument.
authors.affiliation: Division of Plant Biotechnology, College of Agriculture and Life Science, Chonnam National University, Gwangju 500-757, Republic of Korea.
authors.forename: Seunghan
authors.initials: S
authors.lastname: Oh
authors.forename: Dong Hyun
authors.initials: DH
authors.lastname: Kim
authors.forename: Bharat Bhusan
authors.initials: BB
authors.lastname: Patnaik
authors.forename: Yong Hun
authors.initials: YH
authors.lastname: Jo
authors.forename: Mi Young
authors.initials: MY
authors.lastname: Noh
authors.forename: Hyo Jeong
authors.initials: HJ
authors.lastname: Lee
authors.forename: Kwang Ho
authors.initials: KH
authors.lastname: Lee
authors.forename: Kwang Ho
authors.initials: KH
authors.lastname: Yoon
authors.forename: Wan-Jong
authors.initials: WJ
authors.lastname: Kim
authors.forename: Ju Young
authors.initials: JY
authors.lastname: Noh
authors.forename: Heon Cheon
authors.initials: HC
authors.lastname: Jeong
authors.forename: Yong Seok
authors.initials: YS
authors.lastname: Lee
authors.forename: Chuan-Xi
authors.initials: CX
authors.lastname: Zhang
authors.forename: Iksoo
authors.initials: I
authors.lastname: Kim
authors.forename: Yeon Soo
authors.initials: YS
authors.lastname: Han
mesh_terms: D000818:Animals; D002080:Butterflies / Q000821:virology; D005216:Fat Body / Q000821:virology; D016679:Genome, Viral; D025722:Granulovirus / Q000235:genetics* / Q000302:isolation & purification; D007150:Immunohistochemistry; D007814:Larva / Q000821:virology; D008969:Molecular Sequence Data; D000077183:Occlusion Body Matrix Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D015678:Viral Structural Proteins
chemical_list: D000077183:Occlusion Body Matrix Proteins; D015678:Viral Structural Proteins; C000625703:granulin protein, granulovirus