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cDNA cloning and sequence analysis of the gene encoding the peplomer protein of feline infectious peritonitis virus
R J de Groot1, J Maduro, J A Lenstra
1Institute of Virology, Veterinary Faculty, State University, Utrecht, The Netherlands.
Abstract:
The peplomer gene of feline infectious peritonitis virus (FIPV) strain 79-1146 was isolated from a genomic cDNA library by differential hybridization with RNA 2 and 3 as probes. From the nucleotide sequence a primary translation product of 1452 residues (Mr 160,472) was predicted, containing an N-terminal signal sequence, a C-terminal transmembrane segment and 35 potential N-linked glycosylation sites. By S1 nuclease analysis the 5' end of the presumptive RNA 2 body was located at about 30 nucleotides upstream from the initiating AUG codon. At approximately the same position a nine nucleotide sequence ACUAAACUU was found, which was also present 37 nucleotides downstream from the open reading frame. Comparison of the sequences of the FIPV, murine hepatitis virus and infectious bronchitis virus peplomer proteins showed about 27% overall homology, with most conservation in the C-terminal half.
Insights
Researchers isolated the feline infectious peritonitis virus (FIPV) peplomer gene, revealing its protein structure and sequence. This study enhances understanding of FIPV molecular biology and related coronaviruses.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Feline infectious peritonitis virus (FIPV) is a significant pathogen in cats.
- Understanding viral gene structure is crucial for developing control strategies.
Purpose of the Study:
- To isolate and characterize the peplomer gene of FIPV strain 79-1146.
- To predict the protein structure and identify key functional elements.
Main Methods:
- Isolation of the peplomer gene using differential hybridization.
- Nucleotide sequencing to predict the primary translation product.
- S1 nuclease analysis to determine the RNA 5' end.
- Sequence comparison with related viruses.
Main Results:
- A primary translation product of 1452 residues was predicted.
- Identified an N-terminal signal sequence, C-terminal transmembrane segment, and 35 glycosylation sites.
- Located the RNA 2 body and identified conserved sequence motifs.
- Showed approximately 27% homology with murine hepatitis virus and infectious bronchitis virus peplomer proteins.
Conclusions:
- The study provides detailed molecular characterization of the FIPV peplomer gene.
- Identified conserved regions suggest functional importance in related coronaviruses.
- Findings contribute to the broader understanding of coronavirus evolution and pathogenesis.