潮霉素磷酸转移酶的致敏性评价研究

doi:10.7506/spkx1002-6630-200901063

食品科学 ›› 2009, Vol. 30 ›› Issue (1 ): 261-264.doi: 10.7506/spkx1002-6630-200901063

潮霉素磷酸转移酶的致敏性评价研究

许文涛^1,2，芦云²，罗云波²，元延芳²，黄昆仑^1,2,*

1.食品安全分子检测实验室，中国农业大学食品科学与营养工程学院 2.农业部转基因生物食用安全监督测试中心

收稿日期:2007-11-06 出版日期:2009-01-01 发布日期:2010-12-29
通讯作者: 黄昆仑 E-mail:E-mail：hkl009@163.com
基金资助:
国家“863”计划项目(2006AA10Z440)；农业部“948”项目(2007-Z8)

Study on Allergenicity Assessment of hygromycin B phosphotransferase Protein

XU Wen-tao1,2，LU Yun2，LUO Yun-bo2，YUAN Yan-fang2， HUANG Kun-lun1,2,*

(1.Laboratory of Food Safety and Molecular Assessment, College of Food Science and Nutritional Engineering, China Agricultural
University, Beijing 100083, China ；2.Supervision, Inspection and Testing Center of Genetically Modified Food Safety,
Ministry of Agriculture, Beijing 100083, China)

Received:2007-11-06 Online:2009-01-01 Published:2010-12-29
Contact: HUANG Kun-lun1 E-mail:E-mail：hkl009@163.com

摘要/Abstract

摘要：

潮霉素磷酸转移酶基因广泛用于转基因生物的基因转化过程中，该基因(hpt)编码的蛋白属于无食用和食物过敏史的处源目的蛋白。根据IFBC-ILSI 制定的转基因食品致过敏性评价方法，进行了与已知的过敏蛋白的氨基酸序列相似性比较、消化稳定性实验以及动物模型实验。结果表明，未发现连续8 个氨基酸序列相同；该蛋白在80℃加热10min 左右时已全部降解，不具有热稳定性；体外模拟消化实验表明，消化20min 时，SDS-PAGE 和WesternBlotting 已检测不出Hpt 蛋白。实验采用BN 大鼠作为过敏模型，ELISA 检测特异IgG、IgE 以及组胺水平，Hpt 蛋白与阳性存在显著性差异(p ＜ 0.05)，与阴性无显著性差异。综合结果表明，Hpt 蛋白潜在致敏性很低。本研究可为进一步开展以hpt 基因为标记性基因的作物上市前安全性的评价提供基础数据。

关键词: 潮霉素磷酸转移酶, 安全评价, 体外消化, 动物模型, 过敏

Abstract:

Hygromycin B phosphotransferase gene (hpt) has been widely used in the process of gene construction when agricultural plants are modified by biotechnology. Hpt protein as an introduced “new” protein was obtained from sources with no prior history of causing allergy, thus its safety assessment was mainly focused on comparison of amino acid sequences, in vitro digestibility and animal model according to the IFBC-ILSI. Individual sequence analysis with known amino acid sequences, reported as allergens, showed that no eight continuous amino acids are observed; Hpt protein is unstable to heat for its complete decomposition at 80 ℃ for 10 min. It is also digested within the first 20 min by simulated gastric fluid treatment as observed by SDS-PAGE and Western blotting. BN rats were used to establish allergenicity model. The levels of IgG, IgE and histamine were tested by ELISA. The results showed that the Hpt protein has significant difference with the positive (p ＜ 0.05), but no difference with the negative. In conclusion, Hpt protein has low probability to induce allergenicity. The results can provide basic data for evaluating the safety of genetically modified plants which use hpt as marker gene.

Key words: hygromycin B phosphotransferase gene, safety assessment, in vitro digestibility, animal model, allergenicity

中图分类号:

Q555

许文涛1,2，芦云2，罗云波2，元延芳2，黄昆仑1,2,*. 潮霉素磷酸转移酶的致敏性评价研究[J]. 食品科学, 2009, 30(1 ): 261-264.

XU Wen-tao1,2，LU Yun2，LUO Yun-bo2，YUAN Yan-fang2， HUANG Kun-lun1,2,*. Study on Allergenicity Assessment of hygromycin B phosphotransferase Protein[J]. FOOD SCIENCE, 2009, 30(1 ): 261-264.

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潮霉素磷酸转移酶的致敏性评价研究

Study on Allergenicity Assessment of hygromycin B phosphotransferase Protein

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