植物细胞凋亡中死亡蛋白酶基因的克隆及其功能研究

Apoptosis has been discovered in various animal cells and was found to play a fundamental role in normal development,maintenance of homeostasis and pathological processes. To study the mechanism of programmed cell death can provide us not only more facts to understand of life but also a new way to treat diseases. In plants,apoptosis has been reported since 1994 and realized to play a role in some the processes of development and defence mechanisms. With the support of the National Natrue Science Foundation of China,we have done some successful works about apopyosis in plant cells. We have established the model of apoptosis induced by Vk3 , ethylene ,heat shock ,hydroxyl radicals in tobacco and carrot cells, it can be widely used in the fields of PCD research. The reliable cell-free system has been created in our experiments, it provides a useful tool to realize the mechanisms of apoptosis in plants.It is demonstrated by our experiment that death protease also exists in plant cells.An activation of Casp-3-like and Casp-6-like was observed in heat SH-induced apoptosis system of tobacco cells.PARP and lamin-like proteins were cleaved respectively by these death proteases.We discovered that cytochrome b6/f complex could induce apoptosis and cytochrome f may play a important role in plant cells by our experimental results, the further study is in process.

本研究以烟草悬浮细胞为研究模型，分离纯化细胞凋亡过程中起关键作用的死亡蛋白酶，测定其C、N末端的氨基酸序列。然后设计引物通过degenerative PCR以凋亡细胞的mRNA０宸醋伎寺∷劳龅鞍酌富颉Ｔ儆诮湍赶赴鸇Y150中构建其高效表达系统，分离纯化死亡蛋白酶。最后研究其功能，以动物中死亡底物类似物为候选，寻找植物中死亡蛋白酶的底物。..