Cloning, Expression, and Identification of Immunological Activity of an Allergenic Protein in Tartary Buckwheat
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- WANG Zhuanhua
- Institute of Biotechnology, Shanxi University
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- WANG Lan
- Institute of Biotechnology, Shanxi University
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- CHANG Wenjun
- Institute of Biotechnology, Shanxi University
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- LI Yuying
- Institute of Biotechnology, Shanxi University
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- ZHANG Zheng
- Institute of Biotechnology, Shanxi University
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- WIESLANDER Gunilla
- Department of Medical Sciences/Occupational and Environment Medicine, Uppsala University, University Hospital
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- NORBACK Dan
- Department of Medical Sciences/Occupational and Environment Medicine, Uppsala University, University Hospital
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説明
A cDNA fragment encoding a 24 kDa allergenic protein in tartary buckwheat was obtained using reverse transcription PCR, 3′-rapid amplification of cDNA ends (RACE), and nest PCR. The cDNA clone contained 768 nucleotides, including 588 nucleotides in the open reading frame (ORF) and 180 nucleotides in the 3′-terminal sequence. The ORF encoded a functional protein of 195 amino acids. It shared 95% and 93% nucleotide homology with the allergenic storage protein and a legumin-like protein from common buckwheat respectively. The encoding region was expressed in host strain Escherichia coli BL21 (DE3) induced by IPTG at 28 °C. The inclusion bodies of recombinant protein obtained were analyzed by western blot and purified by affinity chromatography. The purity of target protein reached above 95%. After they were refolded by step-wise dialysis, 68% of the inclusion bodies reached soluble state. An analysis of immunological activity showed that the recombinant protein had a specific IgE binding activity. This is the first report of the molecular cloning and expression of the major allergen from tartary buckwheat.
収録刊行物
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 70 (5), 1195-1199, 2006
公益社団法人 日本農芸化学会
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詳細情報 詳細情報について
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- CRID
- 1390001206477017344
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- NII論文ID
- 10018531372
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- NII書誌ID
- AA10824164
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- ISSN
- 13476947
- 09168451
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- NDL書誌ID
- 7926319
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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