水稻ARF基因家族新成员ADP-Ribosylation Factor-Like Protein 5(OsARFL5)的克隆及表达分析

Cloning and Expression Analysis of a Novel Gene OsARFL5 of ARF Gene Family in Rice

小G蛋白家族中的亚家族ADP-核糖基化因子(ADP-ribosylation factor, ARF),在生物体内起众多的生理作用,参与基因表达、细胞骨架重组装、微管的形成以及囊泡和核孔运输等。本研究已成功克隆得到ARF基因家族中的一个新基因OsARFL5,系统进化分析表明该基因与SiARFC1亲缘关系较近;以该基因和其启动子序列构建了两种遗传转化表达载体(pOsARFL5::GUS, pCaMV35S::OsARFL5:EGFP),并获得了相应遗传载体的转基因植株;GUS染色结果表明,该基因在不同时期、不同组织部位表达;洋葱表皮亚细胞定位结果显示,Os ARFL5基因编码的蛋白定位在细胞核。RT-PCR结果显示,OsARFL5在水稻不同生育期的根、茎、叶、叶鞘和穗子中均有不同程度的表达。本研究初步探明了ARF基因家族新成员OsARFL5在水稻生长发育过程中的表达模式。

Abstract ADP-ribosylation factor (ARF)is a sub-family of G protein family and plays important physiological functions in organisms,inclucting gene expression,cytoskeleton recombination,microtubule formation,vesicle and nuclear pore transport.In this study,a novel gene OsA RFL5 belonging to ARF gene family in rice was cloned,and phylogenetic analysis indicated that the gene was closely related to SiARFC1.Two transgenic expression vectors (pOsARFL5::GUS,pCaMV35S::OsARFL5:EGFP)were constructed by using the gene and its promoter sequence,and transgenic plants with corresponding genetic vectors were obtained.The GUS staining results showed that the gene was expressed at different developmental stage and in different tissues.Subcellular localization in onion epidermal cells indicated that the protein encoded by OsARFL5 gene was located in the nucleus.RT-PCR results showed that OsA RFL5 was differently expressed in root,stem,leaf,leaf sheath and head of rice at different growth stages.This study preliminarily demonstrated the expression pattern of OsA RFL5,a new member of the ARF gene family,during the growth and development office.