Characterization of the Transgenic Rice Event TT51-1 and Construction of a Reference Plasmid

被引:18
作者
Cao, Yinglong [1 ]
Wu, Gang [1 ]
Wu, Yuhua [1 ]
Nie, Shujing [1 ]
Zhang, Li [1 ]
Lu, Changming [1 ]
机构
[1] Chinese Acad Agr Sci, Key Lab Oil Crop Biol, Minist Agr, Oilcrops Res Inst, Wuhan 430062, Peoples R China
关键词
TT51-1; characterization; rearrangement; reference plasmid; calibrator; REAL-TIME PCR; GENETICALLY-MODIFIED MAIZE; DNA FRAGMENTS; PLANTS; GENE; QUANTIFICATION; INACTIVATION; QUANTITATION; LIBERTYLINK; VALIDATION;
D O I
10.1021/jf201699s
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
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.
引用
收藏
页码:8550 / 8559
页数:10
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