A RECOMBINANT RICE 16.9-KDA HEAT-SHOCK PROTEIN CAN PROVIDE THERMOPROTECTION IN-VITRO

It is difficult to obtain large amounts of purified low-molecular-mass heat shock proteins (LMM HSPs), which are unique to plants, for bioch emical and physiological studies. Therefore, an attempt was made to pr oduce such a HSP by applying recombinant DNA technology. We fused the cDNA for a rice class I 16.9-kDa HSP, pTS1, to the gene for glutathion e S-transferase (GST) of Schistosoma japonicum and we obtained large a mounts of the fusion protein from transformed Escherichia coli cells, In addition, we found that the 16.9-kDa HSP obtained by cleavage of th e recombinant protein could also form a protein complex of similar to 310 kDa under nondenaturing conditions as can the small, native, class I HSPs from heat-shocked rice seedlings. An assay in vitro to examine the thermoprotection of rice soluble proteins from heat denaturation revealed the strong stabilizing effect of the recombinant HSP.