A shortgun genomic library of the chloroplast DNA (ctDNA) of Hevea brasiliensis (clone RRIM 600) was constructed with the plasmid cloning vector pUC19 in Escherichia coli. The gene, rbcL, for the large sub-unit (LS) of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was isolated by first screening the library by colony hybridisation and then confirmation by Southern hybridization with a radiolabelled DNA probe that contains the internal sequence of rbcL from tobacco, Nicotiana tabacum. Sequence data indicated that the Hevea rbcL gene contains an open reading frame of 1425 nucleotides encoding a protein of 475 amino acid residues. Conserved sequences in the gene and LS protein were detected. Palindromic sequences which may form intrastrand stem-loop structures for the termination of transcription were encountered at the 3 non-coding region of the gene.