检测黄曲霉毒素生物合成相关基因芯片的研制

Preparation of gene chip for detecting different expression genes involved in aflatoxin biosynthesis

目的建立比较成熟的可用于分析黄曲霉毒素生物合成相关基因的芯片技术。方法采用反转录PCR和芯片检测对比的方法。实验所用探针来源于寄生曲霉，待测样品选择黄曲霉菌株。结果芯片点阵后依次通过温浴2h，650mJ／cm^2紫外交联30s，80℃烘烤2h，预杂交45min，清洗，干燥等步骤，最后与待测样品在42℃杂交16h，获得了低背景高质量的芯片。芯片扫描显示荧光信号稳定，与反转录PCR扩增结果一致，并且无背景干扰。结论探针设计合理，实验方法可靠。初步建立了用芯片检测与黄曲霉毒素生物合成相关基因的技术平台。

Objective To develop the methodology of gene chip to analyse genes involved in aflatoxin biosynthesis. Methods In comparing reversed transcriptional PCR with gene chip, the gene chip was used to detect genes involved in aflatoxin biosynthesis. Results After arrayed the slide was incubated in water for 2 hours, exposed to a 650 mJ/cm^2 of ultraviolet irradiation in the strata-linker for 30 s, roasted under 80 ℃ for 2 hours in oven, pre-hybridized for 45 minutes and dealt with other procedures. Finally, the slide was hybridized with fluor-derivatized sample at 42℃ for 16 hours. Conclusion With the reasonable probe design and applicable protocol, the gene chip was prepared effectively for research on genes involved in aflatoxin biosynthesis.