title: Event-specific qualitative and quantitative detection of five genetically modified rice events using a single standard reference molecule.
abstract: One novel standard reference plasmid, namely pUC-RICE5, was constructed as a positive control and calibrator for event-specific qualitative and quantitative detection of genetically modified (GM) rice (Bt63, Kemingdao1, Kefeng6, Kefeng8, and LLRice62). pUC-RICE5 contained fragments of a rice-specific endogenous reference gene (sucrose phosphate synthase) as well as the five GM rice events. An existing qualitative PCR assay approach was modified using pUC-RICE5 to create a quantitative method with limits of detection correlating to approximately 1-10 copies of rice haploid genomes. In this quantitative PCR assay, the square regression coefficients ranged from 0.993 to 1.000. The standard deviation and relative standard deviation values for repeatability ranged from 0.02 to 0.22 and 0.10% to 0.67%, respectively. The Ministry of Food and Drug Safety (Korea) validated the method and the results suggest it could be used routinely to identify five GM rice events.
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Republic of Korea.
authors.forename: Jae-Hwan
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Republic of Korea.
authors.forename: Saet-Byul
authors.initials: SB
authors.lastname: Park
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Republic of Korea.
authors.forename: Hyo-Jeong
authors.initials: HJ
authors.lastname: Roh
authors.affiliation: Food Safety Risk Assessment Division, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of Korea.
authors.forename: Min-Ki
authors.initials: MK
authors.lastname: Shin
authors.affiliation: Food Safety Risk Assessment Division, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of Korea.
authors.forename: Gui-Im
authors.initials: GI
authors.lastname: Moon
authors.affiliation: Food Safety Risk Assessment Division, National Institute of Food and Drug Safety Evaluation, Cheongju 28159, Republic of Korea.
authors.forename: Jin-Hwan
authors.initials: JH
authors.lastname: Hong
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Republic of Korea. Electronic address: hykim@khu.ac.kr.
authors.forename: Hae-Yeong
authors.initials: HY
authors.lastname: Kim
mesh_terms: D018744:DNA, Plant / Q000235:genetics*; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*
chemical_list: D018744:DNA, Plant
keywords: Genetically modified rice; Polymerase chain reaction; Qualification; Quantification; Standard reference plasmid