大豆油酸脱氢酶基因启动子的克隆及其表达活性分析

中国生物工程杂志

2013, Vol. 33

Issue (4): 80-84

研究报告

大豆油酸脱氢酶基因启动子的克隆及其表达活性分析

赵艳¹, 沙伟¹, 张梅娟¹, 杨晓杰¹, 范震宇¹, 王艳梅²

1. 齐齐哈尔大学生命科学与农林学院齐齐哈尔 161000;
2. 齐齐哈尔市龙沙公园齐齐哈尔 161006

Cloning and Activity Analysis of Soybean FAD2-1B Promoter

ZHAO Yan¹, SHA Wei¹, ZHANG Mei-juan¹, YANG Xiao-jie¹, FAN Zhen-yu¹, WANG Yan-mei²

1. College of Life Science and Agroforestry, Qiqihaer University, Qiqihaer 161000, China;
2. LongSha Park of Qigihar City, Qiqihaer 161006, China

全文: PDF(435 KB) HTML

摘要： 大豆油酸脱氢酶(FAD2-1B)基因是种子特异表达基因,利用PCR方法从大豆基因组DNA中分离FAD2-1B 基因的启动子片段,命名为FP。PLACE在线启动子预测工具分析表明:序列中含有多种典型的种子特异性表达元件, 如Skn-1 motif、AACACA、SEF4 motif、E-box、ACGT等。将克隆得到的FP片段替换pCAMBIA1301中的CaMV35S 启动子,构建表达载体pCAM-FP。通过农杆菌介导法在大豆各组织中进行瞬时表达,GUS组织化学染色显示FP驱动GUS基因在大豆根、茎、叶中基本不表达,在种子中有较高的表达活性,推测FP启动子具有种子特异表达活性。

关键词： 大豆; 油酸脱氢酶基因; 启动子; 瞬时表达

Abstract: The soybean oleate desaturase gene(FAD2-1B ) has previously been shown to be expressed specifically in soybean seeds. The 5'-flanking upstream sequence of FAD2-1B gene, named FP, was isolated from the genomic DNA of soybean by PCR method. Sequence analysis by PLACE revealed that this fragment contains a series of motifs related to seed-specific promoters, such as Skn-1 motif,AACACA,SEF4 motif,E-box,ACGT. Replacing CaMV35S promoter of pCAMBIA1301 with FP fragment, the binary expression vector pCAM-FP was constructed. Transient expression in soybean tissues by Agrobacterium tumefaciens mediated method, the results of histochemical GUS analysis showed that there were little or not GUS activities in roots, stems and leaves, but there was higher activity in soybean seeds. It is inferred that FP promoter possess the function driven downstream gene expression exclusively in soybean seeds.

Key words: Soybean FAD2-1B Promoter Transient expression

收稿日期: 2012-10-12 出版日期: 2013-04-25

ZTFLH:

Q786

基金资助: 黑龙江省教育厅科学技术研究资助项目(12521611)

通讯作者: 赵艳 E-mail: zhaoyan3053877@yahoo.com.cn

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引用本文:

赵艳, 沙伟, 张梅娟, 杨晓杰, 范震宇, 王艳梅. 大豆油酸脱氢酶基因启动子的克隆及其表达活性分析[J]. 中国生物工程杂志, 2013, 33(4): 80-84.

ZHAO Yan, SHA Wei, ZHANG Mei-juan, YANG Xiao-jie, FAN Zhen-yu, WANG Yan-mei. Cloning and Activity Analysis of Soybean FAD2-1B Promoter. China Biotechnology, 2013, 33(4): 80-84.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I4/80

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