Yeh C H, Yeh K W, Wu S H, Chang P F, Chen Y M, Lin C Y
Department of Botany, National Taiwan University, Taipei, R.O.C.
Plant Cell Physiol. 1995 Oct;36(7):1341-8.
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.
获取大量纯化的低分子量热休克蛋白（LMM HSPs）很困难，这类蛋白是植物所特有的，难以用于生化和生理学研究。因此，人们尝试应用重组DNA技术来生产这类热休克蛋白。我们将水稻I类16.9 kDa热休克蛋白pTS1的cDNA与日本血吸虫谷胱甘肽S-转移酶（GST）基因融合，并从转化的大肠杆菌细胞中获得了大量融合蛋白。此外，我们发现通过切割重组蛋白获得的16.9 kDa热休克蛋白在非变性条件下也能形成约310 kDa的蛋白复合物，热激水稻幼苗中的小型天然I类热休克蛋白也能如此。一项检测水稻可溶性蛋白热变性热保护作用的体外试验表明，重组热休克蛋白具有很强的稳定作用。
