Expression of the soybean allergenic protein P34 in Escherichia coli and its indirect ELISA detection method

文献类型： 外文期刊

第一作者： Liu, Bin

作者： Liu, Bin;Teng, Da;Wang, Xiumin;Yang, Yalin;Wang, Jianhua;Liu, Bin;Teng, Da;Wang, Xiumin;Yang, Yalin;Wang, Jianhua

作者机构：

关键词： Soybean allergen;P34/Gly m Bd 30K;Prokaryotic expression;pAB;Indirect ELISA

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期： 2012 年 94 卷 5 期

页码：

收录情况： SCI

摘要： To detect the soybean allergen P34 ( Bd 30K) from soybean products, the full-length cDNA sequence of P34 was synthesized and inserted into the prokaryotic expression vector pET-28a. The P34 protein was expressed in BL21 (DE3) as an inclusion body under the induction of 0.8 mmol/L isopropyl beta--1-thiogalactopyranoside. After purification with His-Bind affinity chromatography, the purity quotient of the recombinant protein was over 92 %, and its molecular weight (approximately 33 kDa) was very close to that of the native soybean P34. The polyclonal antibody (pAB) against P34 was prepared with the purified recombinant P34. The generated pAB, named as pAB-P34, exhibited high specificity to the P34 protein of the soybean meal. The indirect enzyme-linked immunosorbent assay (iELISA) based on pAB-P34 was established to determine the P34 content of soybean products. The CVs of the recovery tests of P34 were less than 7.77 %, which indicated that iELISA had high reproducibility and accuracy. Therefore, the recombinant P34 produced in the expression system, the prepared pAB-P34, and the developed iELISA could provide a valuable tool for sensitive detection of P34 in various soybean products and for future studies on allergies related to soybean P34.

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