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Characterization of the transgenic rice event TT51-1 and construction of a reference plasmid
Authors:Cao Yinglong  Wu Gang  Wu Yuhua  Nie Shujing  Zhang Li  Lu Changming
Institution:Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oilcrops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, China.
Abstract:Transgenic rice TT51-1 (BT63) is an insect resistant strain that was granted for safety certificate in China in 2009. This study characterizes the transgenic event TT51-1 using a GenomeWalker strategy. The organization of the transgenes indicated that the transgenes on two plasmids, pFHBT1 and pGL2RC7, had been integrated at the same locus. The sequence of the event TT51-1 spanned 8725 bp, including a truncated Cry1Ab/Ac cassette, an intact Cry1Ab/Ac cassette, two Amp gene segments, and an Hph gene segment. The 5' and 3' plant flanking sequences were isolated and used to locate the transgenes to chromosome 10 in TT51-1. The isolated TT51-1 fragment and a fragment of the rice PLD gene were integrated into a plasmid vector, to create plasmid pK-TT51 as a calibrator for detecting rice containing TT51-1. Analysis of unknown samples indicated that the reference plasmid was a reliable alternative to TT51-1 genomic DNA.
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