Cloning and expression of the coat protein (CP) gene of Apple stem pitting virus (ASPV; Family: Flexiviride, Genus: Foveavirus) using in vivo Escherichia coli and Pichia pastoris as well as in vitro wheat germ extract gene expression system are described. The coat protein gene was amplified for five ASPV isolates representing three groups characterized by a different size of CPs. The amplified RT-PCR products were cloned into bacteria and yeast expression vectors. Plasmid containing the CP gene of the isolate MT32 was used as a template for expression in wheat germ extract. Proteins of expected sizes, reacting with ASPV antiserum in Western blotting experiments, were obtained in three expression systems for the different virus isolates used in this study. The purified proteins were used for immunization and preparation of rabbit polyclonal antisera. The obtained antibodies could be used for immunocap- ture RT-PCR detection of ASPV in woody tissue, but ELISA results were not satisfactory.
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