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Characterization of guinea pig cytomegalovirus DNA
Abstract:
The genome of guinea pig cytomegalovirus (GPCMV) was analyzed and compared with that of human cytomegalovirus (HCMV). GPCMV and HCMV DNAs were isolated from virions and further purified by CsCl centrifugation. Purified GPCMV DNA sedimented as a single peak in a neutral sucrose gradient and was infectious when transfected into guinea pig embryo fibroblast cells. The cytopathology was characteristic of that seen after infection with GPCMV. Virus DNA purified from virions isolated from infected GPEF or 104C1 cells had a CsCl buoyant density of 1.713 g/cm3, which corresponds to a guanine plus cytosine content of 54.1%. The CsCl buoyant density of GPCMV DNA was slightly less than that of HCMV DNA (1.716 g/cm3), but sufficiently different so that the two virus DNA peaks did not coincide. GPCMV DNA cosedimented with T4 DNA in a neutral sucrose gradient. Restriction endonuclease cleavage of GPCMV or HCMV DNAs with HindIII, XbaI, or EcoRI yielded fragments easily separable by agarose gel electrophoresis and ranging from 1.0 X 10(6) to 25.8 X 10(6) daltons. The number, size, and molarity of GPCMV DNA fragments generated by restriction enzymes were determined. Hybridization of restriction endonuclease-cleaved GPCMV DNA with radioactively labeled HCMV DNA and, conversely, hybridization of restriction endonuclease-cleaved HCMV DNA with radioactively labeled GPCMV DNA indicated sequence homology between the two virus DNAs.
Insights
Guinea pig cytomegalovirus (GPCMV) DNA analysis revealed infectious properties and genetic similarities to human cytomegalovirus (HCMV). This comparative genomics study provides insights into GPCMV genome structure and its relationship with HCMV.
Area of Science:
- Virology
- Molecular Biology
- Genomics
Background:
- Human cytomegalovirus (HCMV) is a significant human pathogen.
- Understanding related viruses like guinea pig cytomegalovirus (GPCMV) aids in comparative studies.
- GPCMV serves as a model for studying herpesvirus biology.
Purpose of the Study:
- To analyze the genome of GPCMV.
- To compare GPCMV DNA with HCMV DNA.
- To investigate the genetic relationship and homology between GPCMV and HCMV.
Main Methods:
- DNA isolation from virions and purification using CsCl centrifugation.
- Neutral sucrose gradient sedimentation and transfection into guinea pig embryo fibroblast cells.
- Restriction endonuclease digestion (HindIII, XbaI, EcoRI) and agarose gel electrophoresis.
- DNA-hybridization studies between GPCMV and HCMV.
Main Results:
- GPCMV DNA is infectious and exhibits characteristic cytopathology upon transfection.
- GPCMV DNA has a CsCl buoyant density of 1.713 g/cm³, corresponding to 54.1% guanine-cytosine content.
- Restriction enzyme analysis revealed distinct fragment patterns for GPCMV and HCMV, with sequence homology detected via hybridization.
Conclusions:
- GPCMV DNA possesses infectious properties.
- GPCMV DNA is structurally similar to HCMV DNA, with identifiable sequence homology.
- Comparative analysis deepens the understanding of cytomegalovirus evolution and genetics.