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从柠条锦鸡儿中克隆和功能分析 AmDUF1517 启动子。

Cloning and functional analysis of AmDUF1517 promoter from Ammopiptanthus mongolicus.

机构信息

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Haidian District, Beijing 100081, People's Republic of China.

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Haidian District, Beijing 100081, People's Republic of China.

出版信息

J Biosci Bioeng. 2020 Sep;130(3):233-238. doi: 10.1016/j.jbiosc.2020.04.014. Epub 2020 May 21.

Abstract

Domains of unknown function protein family 1517 (DUF1517) in Ammopiptanthus mongolicus, could be induced by abiotic stresses, whose upstream regulatory sequence might be an ideal source of abiotic-induced promoter. In this study, a 1026-bp promoter of AmDUF1517 from A. mongolicus was cloned. Five deletion fragments (Full, Q1-Q4) of different length of the AmDUF1517 promoter were fused with the β-glucuronidase (GUS) reporter and transformed into Arabidopsis thaliana. The deletion analysis showed that sequences Full, Q1 and Q3 responded well to mannitol, NaCl and 4 °C stresses, while Q2 and Q4 segments did not. The Q3 fragment (280 bp; -280 to -1 bp) showed the highest promoter activity under normal and mannitol, NaCl and 4 °C conditions. The result suggested that Q3 in the AmDUF1517 gene promoter could be a new source of induced promoters for abiotic resistance breeding in plant genetic engineering.

摘要

多结构域未知功能蛋白家族 1517 (DUF1517)在柠条中可被非生物胁迫诱导,其上游调控序列可能是理想的非生物诱导启动子来源。本研究从柠条中克隆了 AmDUF1517 的 1026bp 启动子。将 AmDUF1517 启动子的五个不同长度的缺失片段(全长、Q1-Q4)与β-葡萄糖醛酸酶(GUS)报告基因融合,并转化拟南芥。缺失分析表明,序列全长、Q1 和 Q3 对甘露醇、NaCl 和 4°C 胁迫反应良好,而 Q2 和 Q4 片段则没有反应。在正常和甘露醇、NaCl 和 4°C 条件下,Q3 片段(280bp;-280 到-1bp)表现出最高的启动子活性。结果表明,AmDUF1517 基因启动子中的 Q3 可能是植物遗传工程中用于非生物抗性育种的新诱导启动子来源。

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