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cDNA cloning of the nucleocapsid and nucleocapsid-associated protein genes of mumps virus
Abstract:
cDNA clones of the mumps virus N and P messenger RNAs were isolated from an infected cell cDNA library. The N and P clones selected the two predominant polyadenylated RNAs found in mumps virus-infected cells with mol. wt. of 0.69 X 10(6) and 0.51 X 10(6), respectively. In addition, clones of the P gene hybridized to and selected mRNAs of higher mol. wt. probably representing polycistronic transcripts of the mumps genome. Hybrid-select translation experiments confirmed the specificity of the clones as representing the nucleocapsid (N) and nucleocapsid-associated protein (P) genes.
Insights
Researchers isolated mumps virus nucleocapsid (N) and phosphoprotein (P) genes from infected cells. These clones specifically identified key viral messenger RNAs, confirming their roles in mumps virus replication.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Mumps virus is a significant human pathogen.
- Understanding mumps virus gene expression is crucial for developing antiviral strategies.
- The genetic makeup and transcriptional products of mumps virus require further elucidation.
Purpose of the Study:
- To isolate and characterize complementary DNA (cDNA) clones for the mumps virus N (nucleocapsid) and P (phosphoprotein) genes.
- To identify and analyze the messenger RNAs (mRNAs) transcribed from the mumps virus genome.
- To confirm the functional identity of the isolated cDNA clones through hybrid-select translation.
Main Methods:
- Construction of a cDNA library from mumps virus-infected cells.
- Screening and selection of specific cDNA clones corresponding to N and P mRNAs.
- Hybridization studies using selected clones to identify viral RNAs.
- Hybrid-select translation assays to verify the protein products encoded by the selected mRNAs.
Main Results:
- Isolation of cDNA clones for both mumps virus N and P genes.
- Identification of two predominant polyadenylated mRNAs (0.69 x 10(6) and 0.51 x 10(6) mol. wt.) corresponding to N and P genes.
- Evidence for higher molecular weight mRNAs, likely polycistronic transcripts, selected by P gene clones.
- Confirmation of clone specificity through hybrid-select translation, yielding N and P proteins.
Conclusions:
- The isolated cDNA clones accurately represent the mumps virus N and P genes.
- The study clarifies the mRNA species involved in the expression of N and P proteins.
- Findings provide a foundation for further studies on mumps virus gene regulation and replication.