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SYBR-Green实时荧光PCR检测转基因番木瓜

Real-Time Fluorescent SYBR-Green PCR Detect the Genetically Modified Papaya

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【作者】 刘建越; 邓欣; 康林; 余道坚; 高必达; 陈冬美;

【Author】 LIU Jian-yue1,DENG Xin1,KANG Lin2*,YU Dao-jian2,GAO Bi-da1,CHEN Dong-mei2 (1.College of Bio-Safety Science and Technology,HNAU,Changsha 410128,China;2. Shenzhen Entry-Exit Inspection and Quarantine Bureau,Shenzhen,Guangdong 518010,China)

【机构】 湖南农业大学生物安全科学技术学院; 深圳出入境检验检疫局动植物检疫技术中心; 深圳出入境检验检疫局动植物检疫技术中心 湖南长沙410128; 湖南长沙410128; 广东深圳518010;

【摘要】 分别以RBCL基因和PRSV-CP基因、PRSV-RP基因为内源基因和目标基因,设计了3对特异性引物,对转基因番木瓜进行了SYBRGreen实时荧光PCR检测.结果表明,RBCL基因引物对所有供试样品成功扩增,Ct值为15~16,PRSV-CP引物对转PRSV-CP基因番木瓜样品成功扩增,Ct值为20~25,PRSV-RP引物对转PRSV-RP基因番木瓜样品成功扩增,Ct值为21~22.运用熔解曲线进行产物分析,验证了试验结果的特异性和准确性.

【Abstract】 This thesis studied on the detection of genetically modified papaya by real-time fluorescent SYBR Green PCR using the ribulose bisphosphate carboxylase/oxygenase large subunit as the endogenesis reference gene and the papaya ringspot virus coat protein,replicating protein gene as the target gene.The results showed that RBCL primers can successfully amplify all the experimental samples,Ct value was 15~16;PRSV-CP primers can successfully amplify the experimental samples which contained PRSV-CP gene,Ct value was 20~25;PRSV-RP primers can successfully amplify the experimental samples which contained PRSV-RP gene,Ct value was 21~22.The amplified products were analyzed by the melting sequence allis,it also proved that the experiment results were differential and veracious.

【基金】 深圳出入境检验检疫局项目(SZK03-2002)
  • 【文献出处】 湖南农业大学学报(自然科学版) ,Journal of Hunan Agricultural University(Natural Sciences) , 编辑部邮箱 ,2006年04期
  • 【分类号】S667.9
  • 【被引频次】14
  • 【下载频次】261
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