Genetic transformation of hevea brasiliensis with human atrial natriuretic factor

Agrobacterium mediated genetic transformation was performed on anther callus of hevea cultivar gl1 with a gene encoding human atrial natriuretic factor (hanf),a peptidehormone that is involved in regulating cardiac blood pressure. the gene constructs containing four different lengths of the indigenous hevein promoter,also incorporated the marker nptll for selection of transformants. thus far 28 plants were regenerated and their bud-grafted generations were successfully transferred to the soil; all transformants tested positive for the promoter-transgene inserts. the presence of the hanf transcript in leaf samples was detected by rt-pcr in a number of original transformants and their vegetative generations,and the authenticity of the amplicon was confirmed by nucleotide sequencing. polyclonal anti-atrial natriuretic peptide detected a band in the low molecular weight region in western-immunoblot of leaf protein of an original transformed plant,while malditof-tof and database searches of the corresponding protein band on sds-gel revealed matches to atrial natriuretic peptide in the c-terminus of hanf.