| 甘蓝型油菜种子特异表达油体蛋白启动子PBnOA03的克隆及功能分析 |
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| 引用本文：王海兰，贾庆利，赵翠珠，杨 铮，王 凯，刘香伶，唐 通，宋 欢，张 猛.甘蓝型油菜种子特异表达油体蛋白启动子PBnOA03的克隆及功能分析[J].西北农业学报,2018,27(8):1152~1160 |
| DOI:10.7606/j.issn.1004-1389.2018.08.010 |
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| 基金项目:陕西省国际合作重点项目（2012KW-17）；国家自然科学基金（31270295）。 |
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| 中文摘要:为了改良油菜种子的性状，常常需要研究种子特异表达目的基因的情况。在分析油菜油体蛋白表达模式的基础上，选择种子特异高表达的油体蛋白，根据甘蓝型油菜基因组序列设计引物，利用PCR技术从基因组中克隆到一段715 bp的启动子序列。启动子顺式元件分析表明，这一克隆序列具有CAAT-box和TATA-box 启动子基本元件，另外含有ABA响应元件等多种顺式元件。进一步利用GUS报告基因在拟南芥中对该启动子的功能进行鉴定，结果表明，在转基因株系的幼苗、根、茎、叶、花和种子发育前期及成熟种子中没有检测到GUS基因明显表达，在种子发育的中后期检测到GUS基因的表达逐渐增强。说明这个启动子的表达调控具有一定的种子特异性。 |
| 中文关键词:甘蓝型油菜 油体蛋白 启动子 GUS染色 拟南芥 |
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| Cloning and Function Analysis of Oleosin Promoter PBnOA03 from Brassica napus |
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| Abstract:In order to improve the traits of oilseed,we often need gene specific expression in certain tissues-based on the analysis of expression pattern of oleosins. Based on the analysis of expression pattern of rapaseed oleosins,one seed-specific oleosin gene with higher transcripts level was chosen in this research.Primers were designed according to the genomic sequence of B.napus and a 715 bp promoter sequence was amplified from Brassica napus genomic by PCR.According to promoter online analysis saftwave, this 715 bp promoter sequence contained some promoter basic elements,such as CAAT-box and TATA-box,and also contained a variety of cis elements such as ABA response element.Furthermore,GUS reporter gene was used to identify the function of the promoter in Arabidopsis thaliana.Histochemical staining of GUS activity showed that obvious expression of GUS was not detected in seedling,root,stem,leaf,flower and the seed during early and mature development stages; but high expression levels were found in developing seed at middle and late stages. These results indicate that the promoter of PBnOA03 functions in a seed-specific pattern. |
| keywords:Brassica napus L. Oleosin Promoter GUS staining Arabidopsis thaliana |
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