Transformation of White Clover with a Gene Encoding a White Clover Mosaic Virus Coat Protein

Genes encoding the white clover mosaic virus (WClMV) coat protein . have been introduced into white clover (Trifolimn repens L.) using Agrobacterium tumefaciens mediated T-DNA transfer. Integration of the coat protein gene in regenerated callus has been confirmed hy Southern blotting of polymerase chain reaction (PCR) amplification products generated using specific oligonucleotide primers. A polyclonal antibody, raised against purified WCIMV coal protein, recognised a polypeptide of c. 25 kDa after SOS-PAGE and Western blotting. The recogilised polypeptide co-migrates with the purified coat protein. Using ELISA, levels of coat protein accumulation in transgenic callus have been determined. The scope for using transgenic plants expressing the WClMV coat protein for protection against this plant virus is discussed.