Cloning, Characterization, and Phylogenetic Analysis of a Shrimp White Spot Syndrome Virus Gene That Encodes a Protein Kinase
Resource
Virology 289 (2): 362-377
Journal
Virology
Journal Volume
289
Journal Issue
2
Pages
362-377
Date Issued
2001
Date
2001
Author(s)
Liu, Wang-Jing
Abstract
An open reading frame (ORF) that encodes a 715-amino-acid polypeptide was found in an 8421-bp EcoRI fragment of the shrimp white spot syndrome virus (WSSV) genome. The polypeptide shows significant homology to eukaryotic serine/threonine protein kinase (PK) and contains the major conserved subdomains for eukaryotic protein kinases. Coupled in vitro transcription and translation generated a protein having an apparent molecular mass of about 87 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. For transcriptional analysis of the pk gene, total RNA was isolated from WSSV-infected shrimp at different times after infection. Northern blot analysis with pk-specific riboprobe found a major and a minor transcript of 2.7 and 5.7 kb, respectively. Rapid amplification of the 5′ cDNA ends of the major 2.7-kb pk transcript showed that there were two transcriptional initiation sites located at nucleotide residues -38(G) and -39(G) relative to the ATG translational start codon. Temporal expression analysis by RT-PCR indicated that the transcription of the pk gene started 2 h after infection and continued for at least 60 h. Phylogenetic analysis showed that WSSV protein kinase does not have any close relatives and does not fall into any of the major protein kinase groups. © 2001 Academic Press.
Subjects
Penaeus monodon; Protein kinase; White spot syndrome virus; WSSV pk gene; WSSV Taiwan isolate
Other Subjects
protein kinase; article; DNA virus; gene amplification; genetic transcription; molecular cloning; nonhuman; Northern blotting; open reading frame; phylogeny; polyacrylamide gel electrophoresis; priority journal; protein domain; reverse transcription polymerase chain reaction; RNA transcription; sequence homology; shrimp; transcription initiation; virus gene; White spot syndrome virus; Decapoda (Crustacea); Eukaryota; Monodon; Penaeus monodon; Shrimp white spot syndrome virus
Type
journal article
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