转基因作物虽然可以产生巨大的经济和环境效益，但是，安全性问题一直是转基因生物产业发展的最主要制约因素。国内外现有的转基因生物安全评价内容均没有考虑到外源基因表达的蛋白质是否会与转基因生物的内源蛋白质发生相互作用，并进而导致转基因生物产生非预期效应。本项目以不同生育期转Bt基因水稻为研究对象，首先通过免疫共沉淀技术和蛋白质质谱技术对与外源Bt蛋白发生相互作用的水稻内源蛋白进行大规模挖掘和鉴定，再通过酵母双杂交技术验证互作蛋白的真实性和互作强度，并用激光共聚焦显微镜观察烟草中Bt蛋白与特定水稻内源蛋白的互作，进一步验证互作的真实性。最后通过比较转Bt基因水稻与亲本水稻之间的抗逆性以及转录组和蛋白质组的差异，探索Bt蛋白与特定水稻内源蛋白互作可能产生的非预期生物学效应，为完善转基因生物安全评价的内容提供理论依据。

Transgenic crops can produce huge economic and environmental benefits, however, the safety problem is always the biggest restraining factor for the development of transgenic industry. As the exogenetic proteins expressed by transgenes, whether the transgene proteins interact with the endogenous proteins in transgenic organisms and further lead to unintended biological effects? The existing contents on safety evaluation of transgenic organisms have not taken the issue into account. In the proposed project, the transgenic Bt rice in different growth stages is chosen as the experimental materials to explore whether Bt protein interacts with the endogenous proteins in transgenic rice through the following steps. Step one, the co-immunoprecipitation assay is applied to search the endogenous rice proteins that interact with Bt protein, and then the protein mass spectrometry technology is used to identify the interaction proteins. Step two, the yeast two-hybrid assays are applied to confirm the ineractions between Bt protein and the identified rice proteins and find the interaction degree, and confocal laser scanning microscope is used to further confirm whether Bt protein and the identified rice proteins interact in tobacco leaf cells. Last step, the unexpected biological effects caused by the Bt protein-rice protein interactions will be explored through comprising the difference of the transcriptome and proteome and the stress resistance between the transgenic Bt rice and the parental rice. The project results will provide the theoretical basis for improvement of the biosafety assessment contents of transgenic organisms.