Identification of a collagen-like protein gene from white spot syndrome virus.
The most unique feature of white spot syndrome virus (WSSV) is the presence of a collagen-like protein (termed as WSSV-CLP). In this report, the N-terminal fragment of WSSV-CLP (CLPn) was expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli and purified. Specific antibody was then raised against the purified fusion protein (GST-CLPn). Temporal analysis showed that the WSSV collagen gene was an early viral gene. Immunogold localization using specific antibody revealed that the gold particles, under a transmission electron microscope, were presented along the outer envelope of WSSV virions. This experiment suggested that the collagen gene encoded an envelope protein of WSSV. Using immuno-affinity chromatography, the WSSV-CLP was purified from crudely purified WSSV virions. The WSSV-CLP was N-glycosylated, as indicated by the increased migration in SDS-PAGE after treatment with N-linked glycosidase F.
Li
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Q
Department of Biochemistry and Molecular Biology, School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, PR China.
Chen
Y
Y
Yang
F
F
D000818:Animals; D003400:Astacoidea / Q000821:virology*; D015153:Blotting, Western; D003094:Collagen / Q000737:chemistry*; D004267:DNA Viruses / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics*; D006031:Glycosylation; D008854:Microscopy, Electron; D014158:Transcription, Genetic / Q000235:genetics; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure
D014759:Viral Envelope Proteins; D003094:Collagen