木质部半胱氨酸蛋白酶XCP2(xylem cysteine proteases 2)是植物管状细胞凋亡过程中的自溶酶,参与植物与多种病原物互作,但目前未见对水稻XCP2蛋白酶的报道.此研究克隆获得水稻XCP2编码基因orf序列,经Bgl II和Spe I酶切纯化后,克隆到植物表达载体pCAMBIA1302(含GFP、his tags).将该基因orf序列信息提交至NCBI比对,查找到其特异序列,利用Gateway技术将该特异序列重组到植物RNAi载体pB7GWIWG2(II),并冻融法将重组质粒转化进入土壤农杆菌GV3101.为进一步探究水稻OsXCP2基因在水稻木质部细胞凋亡及水稻与病原物互作中的作用提供条件.%Xylem cysteine proteases 2(XCP2)is the autolytic enzyme in the process of withering and death of plant tracheary cells,which joins in the interactions between plants and many pathogens.However there have not been any reports about rice XCP2 protein.The cds and conserved fragments of rice OsXCP2 were cloned.After digested by restriction endonuclease Bgl II and Spe I,its cds sequence was cloned into plant over-expression vector pCAMBIA1302(contain GFP,his tags).Via Gateway clone technology the conserved frag-ment was reconstituted into plant RNAi vector pB7GWIWG2(II).And for further study of the functions of Rice OsXCP2 gene in the withering and death of the xylem cells of rice and the interaction between rice and patho-gens,the recombinant plasmids were transformed into Agrobacterium GV3101.
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