中文摘要：

ADP核糖基化因子1（ARF1）是Ras超家族的成员,调控细胞内囊泡运输,进而影响细胞的生长发育。为了研究单细胞真核生物八肋游仆虫ARF1的功能,构建了重组表达质粒pET28a-ARF1,在大肠杆菌BL21中进行了表达纯化,获得高纯度的八肋游仆虫腺苷酸核糖基化因子1蛋白EoARF1,随后通过荧光光谱法检测了其与嘌呤核苷酸的相互作用。结果表明,表达纯化的EoARF1蛋白能与嘌呤核苷酸发生相互作用,其结合位点数n约等于1,且与鸟嘌呤核苷酸作用强弱顺序为GTP〉GDP〉GMP,与GDP的结合强度明显大于与ADP的结合强度。

英文摘要：

ADP ribosylation factor 1（ARF1）is a member of the Ras superfamily,which regulate the intracellular vesicle transport and thereby affect the cell growth and development.In order to study the function of ARF1 from Euplotes octocarinatus,a recombinant plasmid pET28a-ARF1 was constructed and expressed in E.coli BL21.High purity of EoARF1 was obtained.The interaction between EoARF1 with purine nucleotides was investigated by using fluorescence spectroscope.The results showed that the EoARF1 protein could interact with purine nucleotides,the binding site value was nearly one and the intensity of the interaction with guanine nucleotides was GTP〉GDP〉GMP.In addition,the binding constant indicated that the interaction between EoARF1 and GDP was stronger than that of ADP.