| 摘要: |
| 【目的】 从大豆中克隆得到β 伴球蛋白α亚基基因的启动子序列7αP，并对其进行功能分析。【方法】 利用PCR技术从大豆基因组DNA中分离β 伴球蛋白α亚基基因启动子序列7αP，将其与GUS基因融合，构建种子特异性表达载体p7αP GUS，通过根癌农杆菌介导法转化烟草(Nicotiana tabacum)NC89，对再生植株进行PCR、Southern blot检测和GUS组织化学分析。【结果】 序列分析表明，7αP长度为1 382 bp，其中含有多种种子特异性启动子的序列元件，如RY重复序列元件、E-box、SEF1-motif、SEF4 motif、（CA）n、Dc3启动子结合因子和ACGT序列元件及一些诱导物应答元件。转基因植株的PCR和Southern blot结果显示，成功地获得了转基因阳性植株；GUS活性检测表明，仅能在种子中检测到GUS活性，而在根、茎和叶等其他组织中均未检测到GUS活性。【结论】 大豆β 伴球蛋白α亚基基因上游1 382 bp片段具有种子特异性启动子功能，7αP为种子特异性启动子。 |
| 关键词: 大豆 种子特异性启动子 功能分析 GUS染色 烟草 |
| DOI： |
| 分类号: |
| 基金项目:国家转基因专项（J99-13-001）；教育部博士点基金项目（20070193005） |
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| Cloning and identification of the seed specific promoter from soybean |
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CAO Bing SHE Xiao ping
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| Abstract: |
| 【Objective】 This paper aimed to clone and identify the seed specific promoter of α subunit gene from soybean.【Method】 In this study, the promoter region of α subunit gene was isolated from the genomic DNA of soybean Jinong 18 by PCR method.The cloned promoter 7αP was fused to the GUS reporter gene to construct plant expression vector p7αP-GUS,which was introduced into Nicotiana tabacum NC89 viaAgrobacterium mediated gene transfer.Analysis of PCR,Southern blot and GUS activities was employed to screen the specific expression. 【Result】 Sequencing analysis showed that the length of the cloned fragment 7αP was 1 382 bp,GenBank accession No.FJ572966.7αP contained all of the motifs,such as RY repeat element,E box，SEF1 motif,SEF4 motif,（CA）n,Dc3 and ACGT motif etc.as well as some inductor responder elements,which constituted the seed specific promoter activity.PCR and Southern blot results showed that the transgenic plants were obtained successfully. GUS activity assays indicated that expression of GUS was active only in seeds,and didn’t exist in the other tissues such as roots stems and leaves.【Conclusion】 The cloned 1 382 bp fragment 7αP of α subunit gene from soybean is seed specific promoter. |
| Key words: soybean seed-specific promoter functional identification GUS staining tobacco |
