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Molecular characterization of two strains of the avian adeno-associated virus (AAAV)
1Institut für Geflügelkrankheiten, Freie Universität Berlin, Federal Republic of Germany.
Abstract:
An avian adeno-associated virus (AAAV) was isolated after propagating a field isolate of the CELO virus (fowl adenovirus serotype 1 (FAV1)) in embryonated eggs. The isolated dependovirus was compared with the known AAAV obtained from the American Type Culture Collection (ATCC VR-865). The genomes were analysed by digestion with several restriction endonucleases. Although both DNAs have the same size, most restriction enzymes produced different restriction patterns. Double digests were used to construct for the first time restriction maps for avian dependoviruses. The two DNAs rendered different restriction maps in which the different restriction sites were mainly located in the middle and right part of the genomes. The effect of these differences on the structure proteins was shown by western blot analysis. In the immunoblot, the immunofluorescence and immunodiffusion test the two dependoviruses were serologically indistinguishable and therefore can be regarded as two different strains of the same virus. To differentiate between both strains we named the original one as AAAV VR-865 compared with the isolated AAAV DA-1.
Insights
A new avian adeno-associated virus (AAAV) strain, AAAV DA-1, was identified and compared to AAAV VR-865. Despite genomic differences, both strains are serologically indistinguishable, indicating they are variants of the same avian virus.
Area of Science:
- Virology
- Molecular Biology
- Genomics
Background:
- Avian adeno-associated viruses (AAAV) are parvoviruses with potential applications in gene therapy.
- A novel AAAV strain was isolated from CELO virus (fowl adenovirus serotype 1) propagation.
Purpose of the Study:
- To characterize and compare a newly isolated AAAV strain (DA-1) with a reference strain (VR-865).
- To construct restriction maps for avian dependoviruses and analyze structural protein differences.
Main Methods:
- Genome analysis using restriction endonuclease digestion and double digests.
- Construction of restriction maps for avian dependoviruses.
- Western blot analysis to assess structural protein variations.
- Serological comparison using immunoblot, immunofluorescence, and immunodiffusion tests.
Main Results:
- Genomic DNA analysis revealed distinct restriction patterns between AAAV DA-1 and AAAV VR-865, despite similar genome sizes.
- Novel restriction maps were generated for both avian dependovirus strains.
- Western blot analysis indicated differences in structural proteins.
- Serological tests demonstrated that both AAAV strains are indistinguishable, classifying them as variants of the same virus.
Conclusions:
- The isolated AAAV DA-1 represents a distinct strain of avian adeno-associated virus.
- Genomic and structural protein differences exist between AAAV strains, but they share common serological characteristics.
- This study provides the first restriction maps for avian dependoviruses, contributing to their molecular characterization.