Molecular cloning, polyclonal antibody preparation, and characterization of a functional iron-related transcription factor IRO2 from Malus xiaojinensis

Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Bibliographische Detailangaben
Veröffentlicht in:Plant physiology and biochemistry : PPB. - 1991. - 67(2013) vom: 20. Juni, Seite 63-70
1. Verfasser: Yin, Lili (VerfasserIn)
Weitere Verfasser: Wang, Yi, Yan, Mudan, Zhang, Xinzhong, Pan, Haifa, Xu, Xuefeng, Han, Zhenhai
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2013
Zugriff auf das übergeordnete Werk:Plant physiology and biochemistry : PPB
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Antibody preparation Expression Iron deficiency Prokaryotic expression Purification Antibodies Plant Proteins Transcription Factors
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245 1 0 |a Molecular cloning, polyclonal antibody preparation, and characterization of a functional iron-related transcription factor IRO2 from Malus xiaojinensis 
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520 |a Transcription factors play important roles in plant growth and responses to environmental stresses. In this study, a novel basic helix-loop-helix iron-related transcription factor, IRO2, containing a 762-bp open reading frame and encoding 253 amino acids, was cloned from the iron-efficient genotype of Malus xiaojinensis. Localization analyses in onion showed that the MxIRO2 protein was targeted to the nucleus and activation studies in yeast indicated MxIRO2-BD had weak transcriptional activation activity. Prokaryotic expression of MxIRO2 in Escherichia coli BL21 (DE3) pLysS cells resulted in high expression levels of the protein when induced with isopropyl-β-d-thiogalactoside. The fusion protein was purified using Ni-NTA His-bind resin, and the purified MxIRO2-His fusion protein was used as the antigen to immunize a New Zealand rabbit. The resulting antiserum was purified by precipitation with 50% saturated ammonium sulfate and DEAE Sephadex A-50 chromatography to obtain the immunoglobulin G fraction. The expression of MxIRO2 in roots and leaves of M. xiaojinensis seedlings under iron deficiency was determined. The results indicated that MxIRO2 was induced in both roots and leaves under iron deficiency. In these experimental conditions, the transcription and translation levels first increased and then decreased under iron deficiency. This work offers an important basis for further investigating the mechanisms of fruit tree adaptation to iron deficiency 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a Antibody preparation 
650 4 |a Expression 
650 4 |a Iron deficiency 
650 4 |a Prokaryotic expression 
650 4 |a Purification 
650 7 |a Antibodies  |2 NLM 
650 7 |a Plant Proteins  |2 NLM 
650 7 |a Transcription Factors  |2 NLM 
700 1 |a Wang, Yi  |e verfasserin  |4 aut 
700 1 |a Yan, Mudan  |e verfasserin  |4 aut 
700 1 |a Zhang, Xinzhong  |e verfasserin  |4 aut 
700 1 |a Pan, Haifa  |e verfasserin  |4 aut 
700 1 |a Xu, Xuefeng  |e verfasserin  |4 aut 
700 1 |a Han, Zhenhai  |e verfasserin  |4 aut 
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773 1 8 |g volume:67  |g year:2013  |g day:20  |g month:06  |g pages:63-70 
856 4 0 |u http://dx.doi.org/10.1016/j.plaphy.2013.02.021  |3 Volltext 
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