Abstract: The complete cDNA of Wheat yellow mosaic virus(WYMV)P1 gene was obtained by RT-PCR from infected wheat leaves. The sequence analysis showed that P1 gene contained 765 nt, encoding a protein of 255 amino acids (aa) with estimated molecular weight of 28.2 kDa. The fragment of P1 ORF was cloned into prokaryotic expression vector pGEX-6P-1. Subsequently, IPTG-induced GST-P1 fusion protein was purified from E.coli BL21 (DE3). The polyclonal antiserum of WYMV P1 was generated in rabbits by immunizing with purified GST-P1 protein. The quality of the resulting antiserum was examined by ELISA. Western blotting analysis showed that P1 was detected from WYMV infected leaves, but not from purified virus preparation. This result indicated that P1 was not packaged into virus particle and worked as a non structure protein.

Key words: Wheat yellow mosaic virus, P1 gene, cloning, prokayotic expression, polyconal antiserum, immunoreaction