食品科学 ›› 2011, Vol. 32 ›› Issue (22): 192-195.doi: 10.7506/spkx1002-6630-201122039

• 分析检测 • 上一篇    下一篇

马铃薯、番茄内源基因PCR检测引物设计及特异性分析

才 华1,栾凤侠2,*,关学佳2,朱 巍3,白 月2   

  1. 1.东北农业大学生命科学学院 2.黑龙江出入境检验检疫局 3.黑龙江省广播电视大学
  • 出版日期:2011-11-25 发布日期:2011-11-11
  • 基金资助:
    国家质量监督检验检疫总局科研项目(2011IK200)

Primer Design and PCR Detection of Potato and Tomato Endogenous Genes

CAI Hua1,LUAN Feng-xia2,*,GUAN Xue-jia2,ZHU Wei3,BAI Yue2   

  1. (1. School of Life Science, Northeast Agricultural University, Harbin 150030, China;2. Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Harbin 150001, China;3. Heilongjiang Radio and Television University, Harbin 150008, China)
  • Online:2011-11-25 Published:2011-11-11

摘要: 对马铃薯UDP-葡萄糖焦磷酸化酶基因(UGPase,U20345)和番茄多聚半乳糖醛酸酶-2a基因(PG-2a,X04583.1)进行序列相似性分析和多序列比对,确定特异片段,设计特异引物,经聚合酶链式反应(polymerase chain reaction,PCR)检测,引物具有较高的特异性;同时建立利用双重PCR检测马铃薯和番茄混合成分的方法,当马铃薯和番茄DNA混合质量为1.25ng时,仍可对其成分进行可靠的鉴定。新设计的马铃薯和番茄内源基因的引物及建立的双重PCR检测方法具有一定的特异性和灵敏性,适于出入境检验检疫部门对马铃薯和番茄制品进行检测。

关键词: 马铃薯, 番茄, 内源基因特异引物, UGPase, PG-2a, 双重PCR

Abstract: The specific fragments of potato pyrophosphorylase gene (UGPase, U20345) and tomato polygalacturonase-2a gene (PG-2a, X04583.1) were identified by BLAST analysis and multiple sequence alignment. Two pairs of primer were subsequently designed and demonstrated high specificity by PCR. A new method was developed to detect potato and tomato product simultaneously by duplex PCR, and 1.25 ng of mixed potato and tomato DNA was successfully detected. With high specificity and sensitivity, this method is applicable for the detection of potato and tomato products by entry-exit inspection and quarantine departments.

Key words: potato, tomato, endogenous gene primer, UGPase, PG-2a, duplex PCR (polymerase chain reaction)

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