基于重测序鉴定耐盐转基因大豆事件外源 T-DNA 整合位点及特异性检测.

[Objectives]In the early stage of this study, we introduced gene BADH into the soybean cultivar ‘Williams 82'to generate the transgenic event L8014, which showed good salt tolerance(1.5% NaCl)and had been cleared for environmental release. Obtaining and analyzing the flanking sequence of T-DNA insertion site were very important for the safety evaluation and application of transgenic events. The characteristics of the transgenic soybean L8014 material were clarified to promote its biosafety evaluation. [Methods]In this study, we detected the copy number of BADH gene by Southern hybridization. Based on genome resequencing technology, Burrows-Wheeler-Alignment(BWA)method was used to compare and analyze the resequencing data with the T-DNA sequence and soybean genome sequence. [Results]T-DNA was found to be in nucleotides 3925017 to 3926022 in the chromosome 10 of soybean genome. Combined with PCR amplification technology, the left and right boundary flanking sequences of exogenous T-DNA integration site were obtained. [Conclusions]Based on these sequences, an event-specific PCR detection method was established, which might provide rapid and accurate detection to identify the transgenic soybean L8014. [ABSTRACT FROM AUTHOR]