番茄、甜椒中转基因成分和内源基因的多重PCR检测方法的建立

食品科学 ›› 2004, Vol. 25 ›› Issue (10): 219-223 .

番茄、甜椒中转基因成分和内源基因的多重PCR检测方法的建立

邵碧英, 江树勋, 陈文炳, 李寿崧

福建出入境检验检疫局技术中心

出版日期:2004-10-15 发布日期:2011-10-24

Development the Multiplex PCR Detection Method of Genetically Modified Components and Inner Gene in Tomato and Sweet Pepper

SHAO Bi-Ying, JIANG Shu-Xun, CHEN Wen-Bing, LI Shou-Song

Fujian Entry-Exit Inspection and Quarantine Bureau

Online:2004-10-15 Published:2011-10-24

摘要/Abstract

摘要： 采用SDS法提取番茄未成熟及成熟果实和甜椒成熟果实中的总DNA,内源rbcL基因扩增结果均为阳性,表明已提取到DNA及DNA中不存在抑制PCR的物质。应用花椰菜花叶病毒(Cauliflower mosaic virus,CaMV)35S启动子、根癌农杆菌胭脂碱合成酶基因(nos)终止子和大肠杆菌K12菌株新霉素磷酸转移酶Ⅱ(nptⅡ)基因的三重PCR检测样品DNA中的转基因成分,结果均为阴性。将番茄、甜椒的DNA和阳性质粒pBI121进行混和,作为PCR反应的DNA模板,成功建立了可同时检测内源rbcL基因、nptⅡ基因、CaMV35S启动子和nos终止子的多重PCR方法。多重PCR方法具有快速、简便、准确等特点,在转基因产品检测上具有重要的应用价值。

关键词: 番茄, 甜椒, 转基因成分, 多重PCR

Abstract: The DNAs of immaturate, maturate tomatoes and sweet peppers were extracted by SDS method. The detection resultsof inner rbcL gene were positive, showing that the DNAs were extracted successfully and there were no substance restrainingpolymerase chain reaction(PCR) in the DNAs. The detection results of Cauliflower mosaic virus(CaMV) 35S promoter,Agrobacterium tumefaciens nopaline synthase(nos) terminator and Escherichia coli strain K12 neomycin phosphotransferaseⅡ(nptⅡ) gene in the tomatoes and sweet peppers with triplex PCR were negative. The multiplex PCR detecting simultaneity innerrbcL gene, nptⅡ gene, CaMV 35S promoter and nos terminator were experimented successfully, with the mixture templet of theDNAs of tomato, sweet pepper and positive plasmid pBI121. The multiplex PCR was quick, simple and practical, and mightplay an important role on the detection of genetically modified food.

Key words: tomato, sweet pepper, genetically modified component, multiplex polymerase chain reaction

邵碧英, 江树勋, 陈文炳, 李寿崧. 番茄、甜椒中转基因成分和内源基因的多重PCR检测方法的建立[J]. 食品科学, 2004, 25(10): 219-223 .

SHAO Bi-Ying, JIANG Shu-Xun, CHEN Wen-Bing, LI Shou-Song. Development the Multiplex PCR Detection Method of Genetically Modified Components and Inner Gene in Tomato and Sweet Pepper[J]. FOOD SCIENCE, 2004, 25(10): 219-223 .

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