紫云英根瘤中特异表达CCPs基因的同源分析及表达定位

将紫云英根瘤中特异表达的编码CCPs(cysteine cluster protein)同源多肽的蛋白AsF259在大肠杆菌表达菌株(DH10B)中进行体外诱导表达,分别检测其在洋葱表皮细胞与紫云英根瘤中的细胞定位,并利用免疫荧光技术,检测紫云英中AsF259基因的时空表达特征。结果表明:AsF259蛋白分布于洋葱表皮的细胞壁、细胞膜、细胞质;基因表达和融合蛋白定位实验显示,AsF259在根瘤中呈特异性表达,在类菌体分化前期大量表达,且AsF259蛋白与类菌体共定位,表明AsF259基因属于NCR家族,在类菌体分化、维持和共生固氮过程发挥重要作用。

Phylogenetic analysis,gene expression and protein localization of a nodule-specific CCPs gene identified in Astragalus sinicus L.

The nodule-specific expressed protein AsF259 which are homologous to the CCPs was identified in Astragalus sinicus,expressed in Escherichia coli (DH10B) to study the subcellular localization in onion epidermal cell and in Astragalus sinicus nodule. The real time qPCR was used to analyze the gene expression. Results showed that AsF259 protein was distributed in the cell wall,cell membrane and cytoplasm,and highly nodule-specific expressed in the nodules with the highest expression level during the crucial stage of bacteroids differentiation. AsF259 protein was co-localized with the bacteroids,indicating that AsF259 gene belongs to a novel member of NCR family and may play an important role in bacteroids differentiation,maintenance and nitrogen fixation.