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[目的]为研究WRKY转录因子在ABA诱导下的表达调控。[方法]首先利用同源克隆法在白菜型油菜中克隆WRKY基因和Actin基因片段,用BLAST和DNAMAN软件对核酸及氨基酸序列进行分析;通过荧光定量PCR技术检测白菜型油菜WRKY基因在ABA处理条件下的相对表达趋势;然后利用相对定量PCR技术(real-timerelative quantificationPCR,以下简称RT-qPCR)检测WRKY基因在ABA(100μmol/L)诱导不同时段的差异表达。[结果]在白菜型油菜中克隆到一段长度为680bp的WRKY基因片段和一段长度为933bp的β-actin基因片段。RT-qPCR实验结果表明,BcWRKY能被ABA诱导表达,且在诱导1h后表达量最高。[结论]成功地克隆了白菜型油菜的WRKY转录因子基因和Actin基因片段,并证明了白菜型油菜WRKY基因的表达受ABA的影响。
[Objective] The research aimed to study the expression regulation of WRKY transcription factor induced by ABA. [Method] The WRKY gene and Actin gene fragment were cloned by homologous cloning method in Brassica campestris. The nucleic acid and amino acid sequence were analyzed by BLAST and DNAMAN software. The fluorescence quantitative PCR was used to detect WRKY gene in Brassica napus L. under ABA treatment conditions Then the relative expression of WRKY gene in different time periods induced by ABA (100μmol / L) was detected by real-time relative quantitative PCR (RT-qPCR). [Result] A 680bp WRKY gene fragment and a 933bp β-actin gene fragment were cloned in Brassica campestris. The result of RT-qPCR indicated that BcWRKY could be induced by ABA and reached the peak at 1 hour after induction. [Conclusion] The WRKY transcription factor gene and Actin gene fragment of Brassica campestris were successfully cloned and it was proved that WRKY gene expression in Brassica campestris was influenced by ABA.