随着近年分子与基因技术的进步，Bt蛋白基因已被转入农作物中并成功表达出了杀虫蛋白。尽管Bt蛋白被认为对于非靶标生物相对安全，但近期研究表明Bt蛋白可通过食物链富集在蜘蛛体内。这就面临一个严峻问题：富集的Bt蛋白在蜘蛛体内是否会抑制或改变基因表达调控酶而导致可能影响蜘蛛生存的其他基因表达的变化？为此我们提出这个计划：（1）使用cDNA microarray技术分析有无Bt蛋白胁迫下蜘蛛全基因组基因表达；（2）运用SAM或RAM统计方法和程序来检测在有无Bt蛋白胁迫下蜘蛛全基因组差异表达的基因；（3）利用DAVID软件对差异表达的基因进行功能分析和信号途径分析；（4）通过蜘蛛毒腺全长cDNA文库构建和EST测序分析，同时利用生物信息学手段，探讨Bt蛋白胁迫对蜘蛛毒素功能基因的影响。该方案的研究成果将有助于我们了解Bt蛋白是否通过食物链富集影响蜘蛛种群，由此为转Bt作物的安全性评价提供科学依据。

Bt toxin is a protein produced by the soil bacterium Bacillus thuringiensis and has been used for over 60 years as a biological pesticide for controlling lepidopteran pests. Due to its protein nature, Bt toxin is unstable and short lived under natural conditions, its use as a biological pesticide has been limited. With recent advances in molecular and transgenic technologies, Bt genes have been genetically engineered into crops and Bt protein successfully expressed for insect pest control. Although Bt proteins are considered relatively safe to non-target organisms, the potential long-term impact of Bt crops on non-target organisms is still a concern. Recent studies indicated that Bt protein could be enriched in the beneficial spider body via the food chain. This raises a serious question: could the enriched Bt protein in spiders inhibit or alter enzyme activities involved in gene expression and regulation, resulting in alternation of expression of other genes that might affect spider survivorship in the Bt rice field? To answer this question, we plan to (1) use cDNA microarray to analyze genome-wide gene expression profiles in two dominant rice field spider species (Pardosa pseudoannulata and Ummeliata insecticeps) between non-Bt and Bt spider groups; (2) to employ the SAM or RAM statistical methods and procedures to identify differentially expressed genes; (3) to identify the functions and signaling pathways of differentially expressed genes using the DAVID software; (4) to investigate the effects of Bt toxin on the genes involved in synthesis of spider toxin by constructing full-length cDNA libraries from the spider venom glands, by sequencing ESTs, and by bioinformatics analysis of ESTs. Results from the proposed research will help us to understand whether or not Bt protein affects spider populations via the food chain enrichment, therefore, providing a scientific basis for the safety evaluation of Bt crops.