title: [Molecular characteristics and specific PCR detection of transgenic rice containing Cry1Ab].
abstract: Bt01 is a new type of rice that has been genetically modified to express Cry1Abprotein. This study confirmed that Cry1Abwas inserted into Bt01 as a single copy using Southern blotting analysis. TAIL-PCR method was further used to obtain its insertion site information. Specific PCR primers and TaqMan probes were designed based on the 5'-integration junction sequence of transgenic rice Bt01. The results showed that the limit of detection (LOD) was ten copies in qualitative PCR. The quantitative PCR assay showed that the LOD was five copies, and the limit of quantification (LOQ) was ten cop-ies. In addition, the accuracy of the established quantitative PCR was verified by detecting two samples containing 3% and 0.5%Bt01, respectively. The quantitative PCR analysis showed the results were 2.7% and 0.47%, respectively.The above results indicated that the event-specific PCR methods developed have high specificity and good sensitivity, which could be effective methods for identifying and testing the genetically modified Bt01 rice.
authors.affiliation: Zhejiang Academy of Agricultural Sciences, Hangzhou, China. yywxf1980@126.com
authors.forename: Xiao-Fu
authors.initials: XF
authors.lastname: Wang
authors.forename: Xiao-Yun
authors.initials: XY
authors.lastname: Chen
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.forename: Huan-Chun
authors.initials: HC
authors.lastname: Zhang
authors.forename: Qing-Mei
authors.initials: QM
authors.lastname: Miao
authors.forename: Jing
authors.initials: J
authors.lastname: Fang
authors.forename: Jun-Feng
authors.initials: JF
authors.lastname: Xu
mesh_terms: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004731:Endotoxins / Q000235:genetics*; D006460:Hemolysin Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis