酵母双杂交技术筛选AtBAG5的下游蛋白-植物发育学专业论文.docxVIP

AbstractPro掣砒咖ed Abstract Pro掣砒咖ed cell death(PCD)plays an indispensable role in development and physiology,but it is unclear whether the mechanisms governing PCD in both plants and animals are similar． Although it is important to note that none of the Core machinery members(c．吕 caspases，Bcl-2 family proteins)have been identified in plant．Other similarities of animal and plant PCD include DNA fragmentation,cell shrinkage,chromatin condensation,cysteine protease activation and generation of reactive oxygen species, indicating the existence ofenmmon elements in botll systems． We hypothesize that plant genomes may have functional equivalents ofBcl-2 family proteins that share tittle or no sequence similarity with animal counterparts． BAG∞CL2一associated athanogene)family proteins wm ofi百nally identified in mammals due to their ability to associate with the antiapoptotic protein,BCL2，剃 promotc cell survival．Further studies revealed that BAG family genes exist in a number of eukaryotes whose members share at least one copy of a 50 amino-acid conserved domain(BAG domain)that interacts、Ⅳitll heat shock protein 70(14SP70／HSC7∞．BAG proteins may serve as bridging molecules that recruit HSP70力}lSC70 to a specific target protein．As a consequence,this family of cochaperones functionally regulates diverse cellular pathways．such雒programmed death(PCD)and sdxess responses． The Arabidopsis BAG gene family shows remarkable structural and functional similarity to its mammalian counterparts．we hypothesize that the Arabidopsia BAG proteins may play a similar function like their animal counterparts in PCD．Then we choice AtBAG5舔bait to inditified proteins interacting with it by yeast two-hybrid． We also have subcloned CDS ofAtBAG5 into pET-32a vector．These plasmids were used to express the recombinant protein in host BL21(DE3)E．cell ceils．Insoluble recombinant protein we∞purified． Seven true positive colomes Wele selected and sequenced,full length sequences were obtained and we searched for