Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oilcrops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, China.
J Agric Food Chem. 2011 Aug 24;59(16):8550-9. doi: 10.1021/jf201699s. Epub 2011 Jul 21.
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.
转Bt 基因抗虫水稻 TT51-1 已于 2009 年获得中国安全证书。本研究采用基因组步移策略对 TT51-1 进行了遗传特征分析。结果表明,两个质粒 pFHBT1 和 pGL2RC7 上的外源基因整合在同一位置,转Bt 基因 TT51-1 的序列大小为 8725bp,包括一个缺失部分序列的 Cry1Ab/Ac 基因、一个完整的 Cry1Ab/Ac 基因、两个 Amp 基因片段和一个 Hph 基因片段。侧翼序列的分离和定位结果显示,外源基因整合在 TT51-1 第 10 号染色体上。将 TT51-1 片段与水稻 PLD 基因片段克隆到质粒载体上,构建了 TT51-1 校准质粒 pK-TT51,可用于检测含 TT51-1 的水稻。用该参考质粒对未知样品进行分析的结果表明,其可作为 TT51-1 基因组 DNA 的替代物。