【摘要】 【目的】利用农杆菌介导法获得了转GbTMEM214基因陆地棉品系，旨在明确转基因棉花株系中T-DNA插入位点的序列特征及检测方法，为其生物安全评价提供依据。【方法】基于基因组重测序技术，利用BLASTn将测序数据与陆地棉TM-1基因组序列进行比对分析，设计特异性引物通过聚合酶链式反应（polymerase chain reaction, PCR）验证插入位点。【结果】携带目标基因GbTMEM214的T-DNA整合在陆地棉基因组D13号染色体57 019 068～57 019 106 bp,T-DNA插入导致受体基因组37 bp的片段缺失；通过PCR扩增获得携带Gb TMEM214基因的T-DNA插入位点的侧翼序列，由此建立了转GbTMEM214基因棉花的特异性检测方法。【结论】基于基因组重测序技术获得了转Gb TMEM214基因棉花的T-DNA插入位点及侧翼序列信息，可为转基因棉花的生物安全评价提供技术参考。更多还原

【Abstract】 [Objective] Gb TMEM214 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.更多还原