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Molecular characterization of a cDNA clone encoding the Epstein-Barr virus (EBV) DNase
1Graduate Institute of Microbiology, College of Medicine, National Taiwan University, Republic of China.
Journal of Virological Methods
|August 1, 1990
Summary
This study identifies a novel Epstein-Barr virus (EBV) DNase gene (BGLF5) and its transcript. The findings reveal sequence variations and confirm the enzyme
Area of Science:
- Molecular Virology
- Genomics
- Biochemistry
Background:
- Epstein-Barr virus (EBV) is a human herpesvirus implicated in various cancers.
- Understanding EBV gene functions, particularly those involved in viral replication and pathogenesis, is crucial.
- The BGLF5 open reading frame (ORF) was previously identified but its function remained uncharacterized.
Purpose of the Study:
- To characterize the BGLF5 open reading frame (ORF) of Epstein-Barr virus (EBV).
- To investigate the enzymatic activity and expression of the protein encoded by the BGLF5 ORF.
- To analyze the transcriptional regulation of the BGLF5 gene in EBV-infected cells.
Main Methods:
- Construction and screening of a cDNA library from EBV-infected P3HR1 cells.
- DNA sequencing of the identified clone (BG9) and comparison with the B95-8 EBV genome.
- In vitro transcription/translation, DNase activity assays, immunoprecipitation, Northern hybridization, and S1 nuclease analysis.
Main Results:
- A cDNA clone, BG9, was isolated and sequenced, corresponding to the BGLF5 ORF of EBV.
- Sequence analysis revealed 14 base differences and 7 amino acid changes compared to published sequences.
- The BG9 clone expressed active EBV DNase, and a 52 kDa protein was immunoprecipitated using nasopharyngeal carcinoma patient serum.
Conclusions:
- The BGLF5 ORF encodes a functional EBV DNase enzyme.
- The identified EBV DNase transcript is approximately 2.0 kb in size.
- Sequence variations in the BGLF5 gene may influence its function or association with disease.