Over-expression of 72 ku protein of wheat yellow mosaic virus inE. coli and preparation of its antiserum

Abstract

By reverse transcription-polymerase chain reaction (RT-PCR), cDNA fragment of wheat yellow mosaic virus (WYMV) RNA2 encoding 72 ku protein has been synthesized and cloned into plasmid pET30a(+) for overexpression in prokaryotic cells. BL21(DE3) pLys S ofE.coli transformed with the recombinant plasmid pETP72 containing the fragment has been induced to express the 72 ku protein on high level. The produced protein has been purified from sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) for its antiserum preparation. In Western-blotting analysis, the antibodies reacted with the 72 ku protein expressed inE.coli.