摘要
[Objective] The aim was to clone the up-regulated expression gene of rice induced by Rhizoctonia solani.[Method] The EST fragment K16 obtained by suppression subtraction hybridization(SSH)was cloned and confirmed by reverse transcription-polymerase chain reaction(RT-PCR).Then RT-PCR products were cloned into the PMD18-T vector and sequenced.The functions of the sequence were predicted with bioinformatics method.[Result] A 1 079 bp gene was obtained.The gene encoded a protein with 236 amino acids.The protein contains many motif sites,two WRKY domains and a C2H2 zinc finger motif.The gene showed high identities with WRKY8,WRKY24 and WRKY30 gene of rice.[Conclusion] The up-regulated expression gene induced by R.solani was representative WRKY family gene.The gene could play an important role on rice sheath blight resistance.
[目的]克隆纹枯病菌诱导的水稻上调表达基因。[方法]对应用抑制差减杂交技术(SSH)获得的纹枯病菌诱导水稻表达的EST序列K16进行电子克隆,根据电子克隆产物设计引物进行RT-PCR、克隆测序,利用生物信息学技术对克隆产物进行功能预测。[结果]克隆出一个长度为1079bp的序列,结构功能分析显示该基因编码236个氨基酸的蛋白,存在多个功能位点,含有2个典型的WRKY结构域和1个C2H2锌指型结构,该基因与水稻WRKY8、WRKY24和WRKY30基因有较高的同源性。[结论]经电子克隆获得的纹枯病菌诱导表达的基因为典型的WRKY家族基因,该基因可能在水稻抗纹枯病中起重要作用。
基金
Supported by Young Academic Backbone Support Program of Heilongjiang Province(1152G022)~~