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Cloning and sequencing of the mumps virus fusion protein gene
Abstract:
The fusion protein (F) gene of mumps virus was cloned from a cDNA library constructed from infected cell mRNA. The F-specific plasmids were identified by hybridization to a degenerate oligonucleotide probe whose sequence was deduced from the N-terminal amino acid sequence of the F2 protein. The complete nucleotide sequence of the F gene was determined. The gene is 1786 nucleotides long and encodes one long open reading frame of 538 amino acids. The F protein has a 19-amino acid signal peptide cleaved between Cys and Val residues. The cleavage site for activation of the F0 protein into the mature F1,2 is Arg-Arg-His-Lys-Arg. A stretch of 30 hydrophobic amino acids near the C-terminus of the protein is followed by several charged amino acids and appears to serve as the anchoring domain for the protein in the lipid bilayer. The F gene of mumps virus is highly related to the F gene of the paramyxovirus SV-5.
Insights
Researchers cloned and sequenced the mumps virus fusion (F) protein gene, revealing its structure and relationship to other paramyxoviruses. This provides insights into viral protein function and evolution.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- The mumps virus fusion (F) protein is crucial for viral entry and cell-to-cell spread.
- Understanding the genetic makeup of the F protein is essential for developing antiviral strategies.
Purpose of the Study:
- To clone and determine the complete nucleotide sequence of the mumps virus fusion (F) protein gene.
- To analyze the deduced amino acid sequence and structural features of the F protein.
Main Methods:
- Construction of a cDNA library from mumps virus-infected cell mRNA.
- Identification of F-specific plasmids using degenerate oligonucleotide probes.
- DNA sequencing to determine the complete nucleotide sequence of the F gene.
Main Results:
- The mumps virus F gene is 1786 nucleotides long, encoding a 538-amino acid protein.
- The F protein possesses a 19-amino acid signal peptide and a specific cleavage site for activation.
- Hydrophobic and charged amino acid domains suggest a membrane anchoring function.
Conclusions:
- The complete nucleotide sequence of the mumps virus F gene has been elucidated.
- Structural analysis reveals key features for F protein function, including signal peptide and membrane anchor.
- The mumps virus F gene shows significant homology to the F gene of paramyxovirus SV-5, indicating evolutionary relatedness.