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 biochemical and physiological studies. Therefore, an attempt was made to produce 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 glutathione S-transferase (GST) of Schistosoma japonicum and we obtained large amounts of the fusion protein from transformed Escherichia coli cells. In addition, we found that the 16.9-kDa HSP obtained by cleavage of the recombinant protein could also form a protein complex of approximately 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.
Yeh
C H
CH
Department of Botany, National Taiwan University, Taipei, R.O.C.
Yeh
K W
KW
Wu
S H
SH
Chang
P F
PF
Chen
Y M
YM
Lin
C Y
CY
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D003001:Cloning, Molecular; D004926:Escherichia coli; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D006358:Hot Temperature; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012995:Solubility
D006360:Heat-Shock Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins