[Objective]GbTMEM214 transgenic Gossypium hirsutum line,obtained using Agrobacterium-mediated method,was used to clarify the sequence characteristics and detection methods of the T-DNA insertion site,and further promote its biosafety evaluation.[Methods]Based on the genome resequencing technology,the sequencing data was compared with the G.hirsutum standard line TM-1 genome sequence by BLASTn,and specific primers were designed to verify the insertion site by polymerase chain reaction(PCR).[Results]The T-DNA carrying the target gene GbTMEM214 was integrated into the position of 57019 068-57 019 106 bp on chromosome D13 of G.hirsutum genome,resulting in 37 bp deletion of cotton genome.Combined with the flanking sequence of T-DNA insertion site obtained by PCR amplification,the specific detection method for GbTMEM214 transgenic cotton was established.[Conclusion]The T-DNA insertion site and flanking sequence of GbTMEM214 transgenic cotton was obtained based on genome resequencing technology,which can provide technical reference for biosafety evaluation of the transgenic cotton.
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