摘要
目的探索适合豆腐样品的DNA抽提方法并采用实时荧光聚合酶链反应(PCR)技术对豆腐试样中的转基因大豆成分进行检测。方法分别采用CTAB和SDS配制抽提缓冲液提取18个豆腐样品中的DNA,针对转基因大豆都含有大豆内源基因lectin及共同元件CaMV35S启动子、nos终止子及epsps基因进行实时荧光PCR扩增。结果 SDS法比CTAB法提取的豆腐DNA质量更好;18个豆腐样品均检测到了lectin,其中有2个样品检测到了CaMV35S启动子的荧光信号,4个样品检测到了nos终止子的荧光信号,所有样品均未扩增出epsps基因。结论 SDS法比CTAB法更适合于抽提豆腐样品的DNA,提取到的DNA可用于实时荧光PCR法检测豆腐内源基因和外源基因。
Objective To explore a better method for the extraction of DNA from tofu sample and investigate whether the tofu containing soybean ingredients. Methods Both CTAB and SDS were used to prepare the extracting buffer for DNA extraction from tofu samples. Gene-specific primers were used according to the fact that transgenic soybeans contain the lectin housekeep gene, the CaMV35S promoter gene, the nos terminator gene and the epsps gene. Real time fluorescence PCR amplification of these genes was performed to detect the presence of these genes. Results The quality of the DNA extracted by the SDS method was better than that of the CTAB method. All 18 tofu samples were detected the presence of the lectin gene, among which, 2 samples were detected the presence of the CaMV35S promoter gene, 4 samples had been detected the presence of the nos terminator gene, but no samples contained the epsps gene. Conclusion The SDS method is more suitable for the extraction of DNA form tofu samples than the CTAB method. Real time fluorescence PCR can effectively detect the presence of the endogenous and foreign genes in transgenic tofu samples.
出处
《食品安全质量检测学报》
CAS
2015年第8期3230-3236,共7页
Journal of Food Safety and Quality
基金
卫生部转基因新品种培育重大专项(2014ZX08011-005)
北京市属高等学校高层次人才引进与培养计划项目(201304177)
首都医科大学"中青年骨干教师"国内交流培养项目~~
关键词
转基因大豆
豆腐
实时荧光聚合酶链反应
transgenic soybean
tofu
real time fluorescence polymerase chain reaction
