摘要： 前期研究中从拟南芥突变体库中筛选获得一株对灰霉病菌侵染表现为感病的突变体。利用TAIL-PCR等技术克隆获得拟南芥抗灰霉病基因T1N6_22。为了进一步明确T1N6_22在拟南芥抗灰霉病过程中的调控机制，利用酵母双杂交技术（Y2H），以构建成功的pAS1/T1N6_22蛋白为诱饵，筛选拟南芥的cDNA文库。通过酵母双杂交筛选，共获得28个酵母克隆。利用T1N6_2基因特异性引物鉴定酵母阳性克隆，并进行测序。共获得了4个可能与T1N6_22互作的蛋白AT2G19480、AT1G06050、AT5G34780和AT1G21400。Blast分析发现，AT2G19480、AT1G06050、AT5G34780和AT1G21400分别为核小体装配蛋白、功能未知蛋白、假定的泛酸还原酶和硫胺二磷酸盐结合折叠超家族蛋白，分别参与到核糖体装配、泛酸盐合成、植物代谢等过程中。研究结果为确定T1N6_22蛋白的互作蛋白，阐明其调控拟南芥抗灰霉病的分子机制奠定了基础。

关键词: 拟南芥, T1N6_22, 酵母双杂交, 互作蛋白

Abstract: Botrytis susceptible mutant named T1N6_22 was achieved from Arabidopsis mutant library.Through the method of TAIL-PCR,resistance-related gene T1N6_22 of Arabidopsis thaliana against Botrytis cinerea infection was cloned.In order to further clarify the regulation mechanism of T1N6_22 gene in Arabidopsis thaliana against Botrytis cinerea infection,vector pAS1/T1N6_22 of T1N6_22 gene was constructed.Taking pAS1/T1N6_22 fusion protein as bait, Arabidopsis cDNA library were screened by the method of yeast two-hybrid system.A total of 28 yeast clones were obtained by yeast two hybrid screening.The positive clones were identified by T1N6_22 gene specific primers and sequenced.Four interacting proteins (AT2G19480,AT1G06050,AT5G34780 and AT1G21400)were obtained by screening Arabidopsis cDNA library.Blast analysis showed that four interacting proteins encoded nucleosome assembly protein,function unknown protein,putative ketopantoate reductase,and oxidoreductase,respectively.These interacting proteins were related to process,biosynthesis of pantothenate,and plant metabolic process.These results would provide foundation for identifying interacting protein of T1N6_22 and clarifying the regulation mechanism of T1N6_22 gene in Arabidopsis resistance.

Key words: Arabidopsis, T1N6_22, Yeast two-hybrid, Interacting protein

中图分类号: 

• Q78

• S432