转基因水稻T-DNA侧翼序列的扩增与分析  

Amplification and Analysis of T-DNA Flanking Sequences in Transgenic Rice

作　　者：方进[1,2] 翟文学[1] 王文明[1] 李素文[3] 朱立煌[1] 

机构地区：[1]中国科学院遗传研究所 [2]北京师范大学,北京100875 [3]北京师范大学

出　　处：《Acta Genetica Sinica》2001年第4期345-351,共7页

基　　金：!国家863高技术项目!(101-01-02-01);国家转基因植物专项!(J99-B-006)

摘　　要：利用现有的转抗白叶枯病基因Ｘａ２１的水稻材料，通过ＴＡＩＬ－ＰＣＲ技术扩增出携带Ｘａ２１基因的Ｔ－ＤＮＡ的侧翼序列。对２４个有效扩增片段的序列分析结果表明，其中１４个侧翼序列是水稻ＤＮＡ，９个含载体主干序列，１个是外源基因Ｘａ２１片段。１４个Ｔ－ＤＮＡ侧翼的水稻ＤＮＡ序列与直接转化法外源基因整合位点的基因组序列具有不同的特点。这些Ｔ－ＤＮＡ在水稻染色体上整合后其两端序列的特点类似于在转基因双子叶植物中观察到的现象。在含主干序列的侧翼序列（３７．５％，９ ／ ２４）中，载体主干序列是以不同的类型出现的。Rice transformation mediated by Agrobacterium tumefaciens has technically matured to some extent, but the mechanics of T-DNA integration in transgenic rice remains largely unknown. Using thermal asymmetric interlaced PCR (TAILPCR), we analyzed the flanking sequences of T-DNAs in transgenic rice plants, in which the resistance gene for rice bacterial blight disease, Xa21, had been integrated stably. Sequence analysis of 24 fragments amplified by TAILPCR showed that of them 14 were rice genomic DNA, 9 contained vector backbone sequences, and one was a fragment of the exogenous gene Xa21. The characteristics of the 14 rice genomic DNA sequences at T-DNA integrated sites are significantly different from those of the reported rice genomic sequences into which exogenous genes were integrated through direct DNA transformation methods. T-DNA border sequences integrated in rice genome have similar features as those integrated in dicotyledonous genomes. In the backbonecontaining flanking sequences (37.5%, 9 ／ 24) vector backbones appeared in different types.

关 键 词：转基因水稻 T-DNA 整合 载体主干序列 扩增 

分 类 号：Q754[生物学—分子生物学]