Simultaneous Detection of Recombinant DNA Segments Introduced into Genetically Modified Crops with Multiplex Ligase Chain Reaction Coupled with Multiplex Polymerase Chain Reaction
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- 作者：Junichi Mano† ; ; Taichi Oguchi† ; ; Hiroshi Akiyama‡ ; ; Reiko Teshima‡ ; ; Akihiro Hino† ; ; Satoshi Furui† ; ; Kaz
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2009
- 出版时间：April 8, 2009
- 年：2009
- 卷：57
- 期：7
- 页码：2640-2646
- 全文大小：223K
- 年卷期：v.57,no.7(April 8, 2009)
- ISSN：1520-5118
文摘
We developed a multiplex polymerase chain reaction (PCR)−multiplex ligase chain reaction (LCR) (MPCR−MLCR) technique as a novel approach for the simultaneous detection of recombinant DNA segments (e.g., promoters, trait genes, and terminators) of genetically modified (GM) crops. With this technique, target DNA regions were amplified by multiplex PCR, the PCR products were then subjected to multiplex LCR as template DNAs, and the LCR products were then analyzed by polyacrylamide gel electrophoresis and subsequent fluorescent scanning. Seven recombinant DNA segments commonly introduced into some GM crop lines were selected as target DNA regions. In addition, another MPCR−MLCR system for the simultaneous detection of three endogenous DNA segments was designed as a positive control test. The specificity and sensitivity of the method were examined. The method allowed us to detect GM crops comprehensively and is expected to be utilized for efficient screening of GM crops into which any one of the seven recombinant DNA segments have been introduced, and for profiling the segments.