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通过血清学和 2D-DIGE 分析研究 EGFP 转基因鸡肉的变应原性。

Allergenicity study of EGFP-transgenic chicken meat by serological and 2D-DIGE analysis.

机构信息

Division of Novel Foods and Immunochemistry, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya, Tokyo, Japan.

出版信息

Food Chem Toxicol. 2010 May;48(5):1302-10. doi: 10.1016/j.fct.2010.02.028. Epub 2010 Feb 25.

Abstract

Genetically modified (GM) foods must be tested for safety, including by allergenicity tests to ensure that they do not contain new allergens or higher concentrations of known allergens than the same non-GM foods. In this study experimentally developed EGFP-transgenic chickens were used and evaluated the allergenicity of meat from the chicken based on a serological and two-dimensional difference gel electrophoresis (2D-DIGE) analysis. For the serological analysis, a Western blotting with allergen-specific antibodies and a proteomic analysis of chicken meat allergens with patients' sera, a so-called allergenome analysis, were used. The allergenome analysis allowed us to identify five IgE-binding proteins in chicken meat, including a known allergen, chicken serum albumin, and no qualitative difference in their expressions between the GM and non-GM chicken meat was found. Results of the 2D-DIGE analysis showed that none of the IgE-binding proteins in chicken meat were significantly changed in expression levels between non-GM and GM chicken, and only 3 of the 1500 soluble protein spots including green fluorescence protein were markedly different as a result of gene transfer. These above results showed that the combination of serological and 2D-DIGE analysis is a valid method of evaluating quality and quantity of allergens in GM foods.

摘要

转基因(GM)食品必须进行安全性测试,包括进行变应原性测试,以确保它们不含有新的变应原或比相同的非转基因食品含有更高浓度的已知变应原。在这项研究中,使用了实验开发的 EGFP 转基因鸡,并根据血清学和二维差异凝胶电泳(2D-DIGE)分析评估鸡肉的变应原性。对于血清学分析,使用过敏原特异性抗体的 Western blot 和患者血清的鸡肉过敏原的蛋白质组学分析,即所谓的过敏原组分析。过敏原组分析允许我们在鸡肉中鉴定出五种 IgE 结合蛋白,包括一种已知的过敏原鸡血清白蛋白,并且在 GM 和非 GM 鸡肉之间没有发现它们的表达存在定性差异。2D-DIGE 分析的结果表明,在非 GM 和 GM 鸡肉之间,鸡肉中没有任何 IgE 结合蛋白的表达水平发生显著变化,只有 1500 个可溶性蛋白点中的 3 个包括绿色荧光蛋白,由于基因转移而明显不同。这些结果表明,血清学和 2D-DIGE 分析的结合是评估转基因食品中过敏原质量和数量的有效方法。

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