水稻OsCYP22互作蛋白的筛选及验证

Screening and validation of OsCYP22 interacting proteins in rice

为了探究水稻亲环素家族基因OsCYP22的功能,构建OsCYP22酵母双杂交诱饵载体,并利用水稻酵母双杂交cDNA文库筛选与OsCYP22相互作用蛋白。自激活试验结果表明,OsCYP22诱饵载体无自激活活性,也对酵母细胞无毒性。通过对酵母双杂交文库的筛选,共得到38个阳性菌落,结合测序结果和生物信息学的方法筛选到20个可能与OsCYP22相互作用的蛋白。进一步对OsCYP22分别与调控植物根系生长相关基因OsCSN5和OsRUB1的全长互作验证,结果显示OsCSN5与OsCYP22在酵母中相互作用。这为深入研究OsCYP22的生物学功能提供了理论依据。

In order to understand the function of cyclophilin family OsCYP22 from Oryza sativa L.,the yeast two hybrid bait vector of OsCYP22 was constructed and proteins interacting with OsCYP22 were screened by yeast two hybrid.The results showed that noself-activating activity and toxicity of OsCYP22 bait vector to yeast cells were detected.,among which 38 posi-tive colonies were obtained by yeast two hybrid screening.Combining sequencing analysis and bioinformatics methods,20 candidates that may interact with OsCYP22 were screened.Further validation of the full-length interactions between OsCYP22 and OsCSN5 and OsRUB1 related to the regulation of plant root growth was conducted.The results showed that OsCSN5 in-teracted with OsCYP22 in yeast.This study provides a theoretical basis for further study of the biological function of OsCYP22.