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Group B rotavirus VP2: sequence analysis, expression, and gene coding assignment
D A Lindsay1, S L Vonderfecht, J J Eiden
1Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218.
Virology
|February 15, 1994
Summary
The VP2 core protein of group B rotavirus (GBR) was identified using gene sequencing and expression in insect cells. This finding aids in understanding GBR structure and developing diagnostics.
Area of Science:
- Virology
- Molecular Biology
- Genomics
Background:
- Group B rotavirus (GBR) causes infectious diarrhea in infants and animals.
- Understanding the genetic makeup of GBR is crucial for developing effective countermeasures.
Purpose of the Study:
- To sequence and characterize gene 2 (IDIRg2) of the infectious diarrhea of infant rats (IDIR) strain of GBR.
- To determine the protein coding assignment of the IDIRg2 gene.
- To analyze the antigenic properties of the expressed protein.
Main Methods:
- Complete sequencing of the IDIRg2 genomic segment.
- Cloning and expression of IDIRg2 in insect cells using a baculovirus vector.
- Homology analysis using BestFit to compare with known rotavirus proteins.
- Antigenicity testing using convalescent antisera and specific antibodies.
Main Results:
- The IDIRg2 gene contains 2847 bp encoding a 934-amino acid polypeptide (106 kDa).
- The deduced amino acid sequence shows limited homology to VP2 of group A rotavirus (SA11 strain).
- Recombinant IDIRg2 protein expressed in insect cells retained antigenic epitopes and reacted with specific antisera.
Conclusions:
- The IDIRg2 gene product is identified as the VP2 core protein of the IDIR GBR strain.
- The expressed VP2 protein is antigenically similar to other GBR strains, including human and porcine isolates.
- This research provides a foundation for further studies on GBR structure and pathogenesis.