表面等离子体谐振(SPR)技术在水稻垩白基因Chalk5检测中的应用

Application of Surface Plasmon Resonance(SPR)?Technique in Detection of Rice Chalk5 Gene

以垩白度低的明恢63与垩白度高的9311的基因组DNA为材料设计探针,构建基于表面等离子体谐振(Surface Plasmon Resonance,SPR)技术的水稻垩白基因检测方法,并用16个已知垩白度的水稻品种检验该方法的可靠性。结果表明:通过生物素修饰后单联DNA片段探针与羧基传感芯片上链霉亲和素(SA)的结合构建基因芯片,再以100μL 0.16μmol/L的PCR产物与探针进行分子杂交,可获得较好的SPR信号;以该方法获得的SPR信号差值的绝对值与水稻垩白成正相关关系,可定性判断水稻品种垩白度的高低;同时,SPR检测方法无需标记,分析过程完全自动化,可实现水稻垩白基因的快速检测。

Using genomic DNA of Minghui 63 with low chalkiness and 9311 with high chalkiness as probes, a method based on Surface Plasmon Resonance（SPR） was developed to detect Chalkiness in rice, and its reliability was tested by 16 rice varieties with known chalkiness. The results showed that single DNA fragment probes modified by biotin combining with streptavidin（SA） on carboxyl sensing chip to construct gene chip, PCR products of rice（100 μL 0.16 μmol/L） were subjected to molecular hybridization and get a more better SPR signal value; The difference of SPR signal obtained by this method is positively correlated with rice chalkiness, and the chalkiness of rice varieties can be qualitatively judged; at the same time, the SPR method did not need markers, and the analysis process was completely automated, which can realize the rapid detection of rice chalkiness gene.