Qualitative and quantitative PCR methods for detection of three lines of genetically modified potatoes.
Qualitative and quantitative polymerase chain reaction (PCR) methods have been developed for the detection of genetically modified (GM) potatoes. The combination of specific primers for amplification of the promoter region of Cry3A gene, potato leafroll virus replicase gene, and potato virus Y coat protein gene allows to identify each line of NewLeaf, NewLeaf Y, and NewLeaf Plus GM potatoes. Multiplex PCR method was also established for the simple and rapid detection of the three lines of GM potato in a mixture sample. For further quantitative detection, the realtime PCR method has been developed. This method features the use of a standard plasmid as a reference molecule. Standard plasmid contains both a specific region of the transgene Cry3A and an endogenous UDP-glucose pyrophosphorylase gene of the potato. The test samples containing 0.5, 1, 3, and 5% GM potatoes were quantified by this method. At the 3.0% level of each line of GM potato, the relative standard deviations ranged from 6.0 to 19.6%. This result shows that the above PCR methods are applicable to detect GM potatoes quantitatively as well as qualitatively.
Rho
Jae Kyun
JK
Plant Biotechnology Division, National Institute of Agricultural Biotechnology, Suwon 441-100, Korea.
Lee
Theresa
T
Jung
Soon-Il
SI
Kim
Tae-San
TS
Park
Yong-Hwan
YH
Kim
Young-Mi
YM
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins; D036022:Capsid Proteins / Q000235:genetics; D018744:DNA, Plant / Q000032:analysis; D004731:Endotoxins / Q000235:genetics; D006460:Hemolysin Proteins; D010942:Plant Viruses / Q000201:enzymology / Q000235:genetics; D030821:Plants, Genetically Modified / Q000145:classification / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017800:Potyvirus / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012680:Sensitivity and Specificity; D011198:Solanum tuberosum / Q000145:classification / Q000235:genetics*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D036022:Capsid Proteins; D018744:DNA, Plant; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis