诱导启动子和病毒5′-非翻译区调控AtDREB2A基因表达提高烟草耐盐性

Q4 Biochemistry, Genetics and Molecular Biology International Journal of Biology and Biomedical Engineering Pub Date : 2021-07-19 DOI:10.46300/91011.2021.15.32
O. Karpova, A. Alexandrova, R. Nargilova, M. Ramazanova, R. Kryldakov, B. Iskakov
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引用次数: 0

摘要

转录因子脱水反应元件结合蛋白2A(DREB2A)在拟南芥的脱水、热休克和盐度中诱导许多基因的表达。缺失编码30个氨基酸的中心区的序列将全长(FL)蛋白DREB2A FL转化为一种更稳定和组成型活性的形式,称为DREB2A CA。在这里,使用农杆菌,通过编码С端带有His标签的蛋白质的转基因AtDREB2A FL-AtDREB2A CA对Nicotiana tabacum v.Samsun NN进行叶盘转化。评估了组成型35S-CaMV启动子或诱导型rd29A启动子与不同的病毒或人工5’-非翻译区(UTR)、5’TMV、5’PVY、5’AMV或5’ARC1(称为翻译增强子)的组合对两种转基因表达的影响。使用His标签的抗体,在正常和热休克条件下在转基因植物中检测到重组蛋白的合成。经过比较分析,不同5’-UTR的性质差异很大,并且依赖于启动子和转基因的单独连接。在rd29A启动子和基因组中的5’TMV或5’AMV的控制下整合AtDREB2A CA有效地提高了烟草转基因植物对400mM NaCl和干旱的耐受性。
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AtDREB2A Gene Expression Under Control of the Inducible Promoter and Virus 5’-untranslated Regions Improves Tolerance to Salinity in Nicotiana Tabacum
Transcriptional factor DEHYDRATION-RESPONSIVE ELEMENT BINDING PROTEIN 2A (DREB2A) induces the expression of many genes in dehydration, heat shock, and salinity in Arabidopsis. Deletion of sequence coding the 30 amino acid central region transforms full length (FL) protein DREB2A FL into a more stable and constitutively active form known as DREB2A CA. Here, using agrobacteria, a leaf disc transformation of Nicotiana tabacum v. Samsun NN was carried out by transgenes AtDREB2A-FL and AtDREB2A-CA coding the proteins with His-tag on the С-end. The effects of combinations of constitutive 35S CaMV promoter or inducible rd29A promoter with different viral or artificial 5’-untranslated regions (UTR), 5’TMV, 5’PVY, 5’AMV or 5’ARC1, known as translational enhancers were evaluated on the both transgenes’ expression. Using an antibody to His-tag, recombinant protein synthesis was detected in transgenic plants in normal and heat shock conditions. After comparative analysis, it was shown that the properties of different 5’-UTRs vary greatly and depended on separate conjunction of promoter and transgene. The integration of AtDREB2A CA under control of the rd29A promoter and 5’TMV or 5’AMV in genome effectively improved tolerance of tobacco transgenic plants to 400 mM NaCl and to drought.
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来源期刊
International Journal of Biology and Biomedical Engineering
International Journal of Biology and Biomedical Engineering Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
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期刊介绍: Topics: Molecular Dynamics, Biochemistry, Biophysics, Quantum Chemistry, Molecular Biology, Cell Biology, Immunology, Neurophysiology, Genetics, Population Dynamics, Dynamics of Diseases, Bioecology, Epidemiology, Social Dynamics, PhotoBiology, PhotoChemistry, Plant Biology, Microbiology, Immunology, Bioinformatics, Signal Transduction, Environmental Systems, Psychological and Cognitive Systems, Pattern Formation, Evolution, Game Theory and Adaptive Dynamics, Bioengineering, Biotechnolgies, Medical Imaging, Medical Signal Processing, Feedback Control in Biology and Chemistry, Fluid Mechanics and Applications in Biomedicine, Space Medicine and Biology, Nuclear Biology and Medicine.
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