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拟南芥抗致病疫霉突变体T-DNA 侧翼序列分析
引用本文:毛娜,刘颖超,董金皋.拟南芥抗致病疫霉突变体T-DNA 侧翼序列分析[J].农业生物技术学报,2007,15(6).
作者姓名:毛娜  刘颖超  董金皋
作者单位:河北农业大学
摘    要:本文通过将致病疫霉不同菌株接种到拟南芥Col-0生态型激活标签突变体上,筛选出了感致病疫霉的类型,以筛选出的拟南芥感病突变体基因组DNA为模板,利用热不对称交错PCR方法(Thermal asymmetric interlaced PCR,TAIL-PCR),进行了抗性相关基因的研究。通过TAIL-PCR技术,获得了拟南芥T-DNA插入侧翼序列,利用拟南芥基因组全测序的优势,通过NCBI进行序列的同源性比对找到相应的基因。结果发现,TAIL-PCR可以有效地扩增T-DNA插入侧翼拟南芥基因组序列,AD1,AD2,和AD4是最佳随机引物。对8个TAIL-PCR特异产物测序分析,证明3个T-DNA插入在基因上。

关 键 词:拟南芥  致病疫霉  激活标签  TAIL-PCR
收稿时间:2006-10-26
修稿时间:2007-1-9

Analysis T-DNA flanking sequence of Arabidopsis thaliana mutants of resistant to Phytophthora infestans
Abstract:Several Phytophthora infestans-susceptible mutants had been obtained based on the different reactions of Arabidopsis activated tagging mutants after being inoculated by the fungus, and flanking sequence of Arabidopsis T-DNA insertion had also been obtained by TAIL-PCR technology and genomic DNA from disease-susceptible mutants as template. Some disease resistant-related genes had been found using NCBI sequence homology analysis and AD1,AD2 and AD4 were the most efficient arbitary primers. Two mutants had been found with T-DNA insertion through sequencing and analyzing eight specific TAIL-PCR products.
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