Qualitative and quantitative analysis of genetically modified pepper.
For the development of qualitative and quantitative PCR methods of genetically modified (GM) pepper developed in Korea, a capsanthin-capsorubin synthase (CCS) gene was used as the endogenous reference gene. The primer pair ccs-F/R amplifying the pepper endogenous gene gave rise to an amplicon of 102 bp. No amplified product was observed when DNA samples from 16 different plants were used as templates. The construct-specific primer pairs amplifying the junction region of the bar gene and Ti7 introduced in GM pepper gave rise to an amplicon of 182 bp. Quantitative PCR assay was performed using a TaqMan probe and a standard plasmid as a reference molecule, which contained both an endogenous and event-specific sequence. For the validation of this method, the test samples containing 0.1, 1, 3, 5, and 10% GM pepper were quantified.
Institute of Life Science & Resources and Graduate School of Biotechnology, Kyung Hee University, Suwon 446-701, Korea.
Hee-Sung
HS
Song
Jae-Hwan
JH
Kim
Dong-Hern
DH
Kim
Hae-Yeong
HY
Kim
D001483:Base Sequence; D015139:Blotting, Southern; D002212:Capsicum / Q000235:genetics*; D018628:Gene Dosage; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
D010940:Plant Proteins; D010088:Oxidoreductases; C086535:CCS protein, Capsicum annuum