Along with transgenic technology in nuclear genome, chloroplast transformation technology being applied to the plant is capable of producing recombinant plant-derived pharmaceutical proteins. The chloroplast transformation of the tomato leaves and cotyledons with the HIV-1-p24 gene was successfully performed using the biolistic method. The bombarded tissues were selected on the callus induction medium MS vitamin B5 1 mg/l BA, 0.1 mg/l NAA supplemented with the spetinomycin from 50 mg/l to 300 mg/I through many selection cycles to be able to get the homoplasmic callus. Tomato plants regenerated from the homoplasmic callus in MS medium supplemented with vitamin B5 2.0 mg/l zeatin, 0.2 mg/l IAA, 300 mg/l spectinomycin. PCR and Southern blot results showed that HIV-1-p24 gene was successfully integrated into tomato chloroplast genomes in 4 transplastomic tomato plant lines. Of four transplastomic lines, three were derived from leaf transformation and one derived from cotyledon transformation. Western blot and ELISA results indicated the presence and the expression of the recombinant protein HIV-1-p24 in the transplastomic tomato lines with an average concentration of 2.35 percent. Thus, complete the process of chloroplast transformation tomato by biolistic and lines of homo plasmic tomato plant expressing of HIV-1-p24 gene.