Isolation and Characterization of a Novel Seed-Specific Promoter from Malaytea Scurfpea
Isolation and Characterization of a Novel Seed-Specific Promoter from Malaytea Scurfpea
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DOI：
10.1007/s11105-014-0785-2
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发表时间：
2015-08
影响因子：
2.1
通讯作者：
Xiao-Ping Fang;D. Kang;Chengjian Xie;Xingyong Yang;Anping Sui
中科院分区：
文献类型：
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作者：
Xiao-Ping Fang;D. Kang;Chengjian Xie;Xingyong Yang;Anping Sui
PSC-AFP是从中草药补骨脂中分离得到的一种具有胰酶抑制活性的抗真菌蛋白，该蛋白在种子中优先表达。用染色体步移法从编码该蛋白的基因的ATG起始密码子位置分离到一个737个碱基的上游序列，鉴定为PPSC-AFP。对PlantCare数据库中的顺式作用元件的分析表明，PPSC-AFP含有RY元件和Gcn4基序，这是一种种子特有的元件，还有一个SKn-1基序。将该片段融合到β-GUS报告基因的上游，利用叶盘法将其转化烟草，研究其功能。GUS组织化学染色确定其表达模式。该片段在种子中表现出较强的GUS活性，而在根、茎和生殖器官(包括雄蕊和雌蕊)中的表达水平较低，表明PPSC-AFP能够以种子特异的方式指导GUS在转基因烟草中的表达。
Psc-AFP, an antifungal protein with trypsin inhibitor activity, was isolated from a traditional Chinese herb, malaytea scurfpea (Psoralea corylifoliaL.); the protein was found to be preferentially expressed in seeds. A 737-bp upstream sequence from the ATG initiation codon site of the gene encoding this protein was isolated using a chromosomal walking method and identified as pPsc-AFP. The analysis of thecis-acting elements in PlantCARE database indicated that pPsc-AFPcontains RY elements and GCN4 motifs, a kind of seed-specific elements, and a Skn-1 motif. The features of this fragment were studied by fusing it into the upstream region of theβ-glucuronidase (GUS) reporter gene and transforming the recombinant expression vector pBI121-pPsc-AFP::GUS into tobacco by using the leaf disk method. The expression pattern was determined using GUS histochemical staining. The fragment showed a strong GUS activity in seeds, whereas its expression level was considerably lower in other organs such as the roots, stems, and reproductive organs, including stamens and pistil, indicating that pPsc-AFPwas able to direct GUS expression in a seed-specific manner in transgenic tobacco.