摘要
根据转基因玉米中所转入的外源基因,选择CaMV35S启动子、NOS终止子、PAT基因和目的基因IVS2/PAT、CDPK/CryIA(b)、Maize genome/CaMV35S、PAT/CaMV35S!、Cry9C/CaMV35S!以及内源IVR基因设计特异性引物,采用多重PCR法对待测样品进行扩增,通过缺口平移法合成DIG-dUTP标记杂交探针,并制备基因芯片。在对PCR反应和扩增产物与芯片杂交条件进行优化的同时,比较了芯片检测的特异性和重复性,并对检测的灵敏度进行测试。结果表明,该方法具有较好的特异性和重复性,检测灵敏度可达0.1%。
According to the plasmid map of Genetically Modified Maize, we select exogenous genes（CaMV35S promoter, NOS terminator, PAT gene, IVS2/PAT, CDPK/CryIA （b）, Maize genome/CaMV35S, PAT/CaMV35S! , Cry9C/CaMV35S! ） and endogenous gene （IVR） as target genes, design and synthesize their primers. The probe was synthesized using the method of nick translation and labeled with DIG-dUTP. Multiplex PCR was used to amplify the target sequence in maize sample DNA, then we hybridized the DNA chips with PCR product, at last the chips displayed the hybridization result. The DNA chip for identifying Genetically Modified Maize was highly specific and repeatable and its sensitivity was 0.1%.
出处
《食品研究与开发》
CAS
北大核心
2009年第6期149-153,共5页
Food Research and Development
