中文摘要：

传统的纯化外源基因的方法主要是通过多代自交,该方法耗时长,效率低。对转基因材料中外源基因T-DNA区域插入位点两翼序列的分析和扩增,是在转基因后代中筛选纯合转基因植株的新方法。首先,利用酶切连接接头的PCR方法,在番茄（Lycopersicon esculentum）转基因植株中扩增外源基因Avr3a的两翼序列并测序;然后通过该序列设计的引物在转基因植株中验证,得到的两翼序列通过与SGN数据库比对分析,结果发现,外源基因的插入位点有40bp的碱基缺失;最后利用侧翼序列和边界序列设计的引物,在转基因T1代植株中筛选出纯合单株,这些植株在T2代进行验证确实为纯合单株。

英文摘要：

Traditional method of screening homozygous transgenic plants is several generations self-pollenation, it is a time consuming work and low efficiency.A new method by flanking sequences amplification of T-DNA insertion site to screen homozygous plants was created.Firstly, adaptor ligation PCR method amplified flanking sequences of T-DNA in tomato（Lycopersicon esculentum）transgenic plants.Then flanking sequences were confirmed by PCR analysis in transgenic plants.BLAST results in SGN database showed that T-DNA integration site lost 40 bp.Finally, homozygous lines were screened out in T1 transgenic generation using flanking sequence primers and border sequence primers, and verified the results in T2 generation.