【摘要】 为建立一种快速检测转基因牛的多重PCR方法,针对牛线粒体16 S rRNA基因(BOS mtD-NA16 S rRNA,BOS)、转基因动物常用标记基因新霉素磷酸转移酶基因(NPTⅡ)、人乳铁蛋白编码基因(humanlactoferrin gene,hLF)、人-α-乳清蛋白编码基因(humanα-lactalbumin gene,hα-LALBA)和人溶菌酶编码基因(humanlysozyme gene,hLYZ)设计了5对特异性引物,通过对扩增条件的优化,建立转基因牛的多重PCR检测方法。利用人乳铁蛋白转基因牛、人溶菌酶转基因牛、人α-乳清蛋白转基因牛样品和其他转基因动物样品进行了敏感性和特异性试验。结果表明方法具有较好的特异性和敏感性,与转人防御素3基因奶牛样品和转omega-3基因猪样品等均无交叉反应,最低检出限达1 ng,该方法既可筛查转基因牛样品,又可进一步确认转基因牛外源基因,适合转基因牛的快速检测。更多还原

【Abstract】 A novel multiple PCR assay was established for detection of genetically modified bevine.Five pairs of specific primers were designed based on the sequences of neomycin phosphotransferase(NPTⅡ),bovine mtDNA 16 S rRNA gene(BOS mtDNA16 S rRNA,BOS),human lactoferrin gene(hLF),human α-lactalbumin gene(hα-LALBA) and human lysozyme gene(hLYZ).The sensitivity and specificity of the method were evaluated with recombinant human lactoferrin transgenic cloned bovine samples,recombinant huma α-LALBA cloned bovine samples,recombinant human lysozyme cloned bovine samples and other genetically modified animal samples.The results showed that the method was very specific and sensitive.It’s dectction limit is 1ng.The method was optimized for not only screening genetically modified bovine but also for identifying the exogenous gene,and it is useful for the transgenic bovine detection.更多还原