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植物研究 ›› 2020, Vol. 40 ›› Issue (6): 932-942.doi: 10.7525/j.issn.1673-5102.2020.06.016

• 研究报告 • 上一篇    

杂交构树UDP-葡萄糖脱氢酶基因编码蛋白的亚细胞定位及其启动子5'端缺失片段的功能分析

吉仁花1, 张文波1, 林晓飞2, 包颖亮1, 特日格勒1, 包会嘎3, 白淑兰1()   

  1. 1.内蒙古农业大学林学院,呼和浩特 010019
    2.内蒙古大学生命科学学院,呼和浩特 010021
    3.乌海市湿地管理局,乌海 016000
  • 收稿日期:2019-12-17 出版日期:2020-11-20 发布日期:2020-11-04
  • 通讯作者: 白淑兰 E-mail:baishulan2004@163.com
  • 作者简介:吉仁花(1986—),女,博士,主要研究方向为林木生物技术。
  • 基金资助:
    国家自然科学基金(31660213);内蒙古自治区研究生科研创新基金(B20171012913)

Subcellular Localization of the Protein Coded by the UDP-Glucose Dehydrogenase Gene from Paper Mulberry and Functional of Its Promoter 5′-end Deletion Fragment

Ren-Hua JI1, Wen-Bo ZHANG1, Xiao-Fei LIN2, Ying-Liang BAO1, Ri-Ge-Le TE1, Hui-Ga BAO3, Shu-Lan BAI1()   

  1. 1.Forestry College,Inner Mongolia Agricultural University,Hohhot 010019
    2.College of Life Science,Inner Mongolia University,Hohhot 010021
    3.Wetlands Authority of Wuhai,Wuhai 016000
  • Received:2019-12-17 Online:2020-11-20 Published:2020-11-04
  • Contact: Shu-Lan BAI E-mail:baishulan2004@163.com
  • About author:JI Ren-Hua(1986—),female,Ph.D.,majoring in the forest biotechnology.
  • Supported by:
    National Natural Science Foundation of China(31660213);Autonomous Region Graduate Research Innovation Fundation of Inner Mongolia(B20171012913)

摘要:

为了研究杂交构树UDP-葡萄糖脱氢酶基因(DDBJ,BpUGDH基因登录号为LC457701)启动子不同区域的表达活性,利用5'端缺失及同源重组实验技术,将5个不同长度的BpUGDH启动子5'端缺失片段与GUS基因连接,并通过农杆菌介导法瞬时转化烟草;同时,为了定位BpUGDH基因编码的蛋白在细胞中表达的具体位置,利用GFP报告基因融合目的基因进行蛋白质的亚细胞定位。结果显示:BpUGDH基因启动子-244 bp以内的序列均能介导GUS基因的诱导表达,并且-973、-465、-355、-281和-244 bp之间的区域可能对BpUGDH基因启动子的活性发挥着至关重要的作用。另外,BpUGDH基因编码蛋白的亚细胞定位结果显示:BpUGDH位于叶绿体中。

关键词: 顺式作用元件, BpUGDH, 亚细胞定位, 杂交构树, 5'端缺失分析法, 瞬时表达

Abstract:

We analyzed the function of cis-acting elements of UDP-Glucose Dehydrogenase Gene from Paper Mulberry(DDBJ, BpUGDH accession No.LC457701)promoter, the five different lengths of BpUGDH promoter 5'-terminal deletion fragment were ligated with the GUS gene by using 5'terminal deletion and homologous recombination techniques, and conducted through heterologous expression in Nicotiana benthamiana by agro-infiltration method. In order to locate the specific location of the protein encoded by BpUGDH gene in cells, subcellular localization of BpUGDH protein was carried out by using the fusion target gene of GFP reporter gene. The results show that the sequences within -244 bp can mediate the induction of GUS gene expression, and the region between -973, -465, -355, -281, -244 bp of the BpUGDH promoter were crucial for regulating the promoter activity. Furthermore, BpUGDH was located in chloroplast.

Key words: Cis-acting element, BpUGDH, subcellular localization, paper mulberry, 5'-end deletion analysis method, transient expression

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