热带作物学报2011，Vol.32Issue(8)：1527-1531,5.DOI:10.3969/j.issn.1000-2561.2011.08.029
转基因大豆插入位点分析及特异性PCR检测方法的建立
Flanking Sequence Analysis and Qualitative PCR Detection of Transgenic Soybean
摘要
Abstract
This paper was designated to discover the integration site of the transgene in the soybean genome and to establish event specific methods for qualitative detection of LEC\ based on the left border junction fragment which was isolated using the TAIL-PCR. Designing the degenerate primers and specific primers based on T-DNA flanking sequence of over-expression vector, PCR to obtain sequences of four homologous. To get Blastn the sequences, analysis three of them: pl48, p225, p69, are both as the carrier T-DNA segment part, without the soybean genome sequence; the rest P217 shows that the sequence is length of 863 bp, Which contains 720 bp fragment of the soybean genome, of which was homologous 99% with soybean photosystem II thylakoid membrane protein of the 206 -926 sections. This suggests that exogenous DNA into the 206 of genome of soybean thylakoid membrane protein coding region. During the integration of the construct LEC event- specific, qualitative PCR method was established with the primers(LECl-s)targeting the junction regions to produce a 277 bp product The method developed in this study proved to be highly specific, sensitive, and suitable for LEC1 sample detection.关键词
转基因大豆:LEC1基因/事件特异性检测/TAIL-PCRKey words
Soybean/ LECl/ Event- specific detection Qualitative PCR/ TAIWCR分类
农业科技引用本文复制引用
郭娜娜,吴辉,于晓惠,黄惜,陈银华,彭明..转基因大豆插入位点分析及特异性PCR检测方法的建立[J].热带作物学报,2011,32(8):1527-1531,5.基金项目
转基因重大专项课题(No.2008ZX08012-001). （No.2008ZX08012-001）
