Sequence and phylogenetic analysis of P10- and P17-encoding genes of avian reovirus.

Avian reovirus (ARV) causes viral arthritis, chronic respiratory diseases, and malabsorption syndrome. The P10 protein is a viroporin and induces cell fusion, whereas the biological function of P17 protein is completely unknown. In this study, the nucleotide sequences of the P10- and P17-encoding genes from 17 field isolates and vaccine strains of ARV isolated over a 23-year period from distinct geographic locations were analyzed to define phylogenetic profiles and to study sequence variability and genetic evolution.

Secreted expression of the VP2 protein of very virulent infectious bursal disease virus in the methylotrophic yeast Pichia pastoris.

The VP2-encoding gene of very virulent infectious bursal disease virus (vvIBDV) was amplified using reverse transcription (RT)-polymerase chain reaction (PCR) and inserted into pPICZalphaA vector. Recombinant plasmid DNA was integrated into the chromosome of the transformed Pichia pastoris by electroporation and expressed protein identified by SDS-PAGE and Western blotting. High-level secreted expression was performed by determining the Mut+ phenotype and secreting multi-copy integrants in the recombinant yeast.

Rapid characterization of avian reoviruses using phylogenetic analysis, reverse transcription-polymerase chain reaction and restriction enzyme fragment length polymorphism.

A reverse transcription-polymerase chain reaction is described, which amplified the full-length sigmaC-encoding and sigmaNS-encoding genes of avian reovirus (ARV). DNA fragments of 1022 and 1152 base pairs were amplified among ARV isolates, respectively, indicating that there were no apparent deletions or insertions in these regions. Fragments amplified from vaccine strains and field isolates were digested with five different restriction enzymes Bcn I, Hae III, Taq I, Dde I, and Hinc II, respectively.