| 摘要: |
| 【目的】克隆小麦蒜氨酸酶基因CDs区，并对其进行序列特征分析和实时定量表达检测。【方法】采用电子延伸结合RT-PCR方法，从小麦叶片分离出蒜氨酸酶基因，网络资源分析其序列特征，实时定量PCR技术检测其在不同条件下的表达情况。【结果】克隆到1个小麦蒜氨酸酶基因TaAly1，其开放阅读框全长1 023 bp，编码340个氨基酸，与水稻和玉米蒜氨酸酶基因的同源性为70%，其基因组DNA序列含有3个内含子；TaAly1在小麦幼苗根部几乎不表达，在叶部表达量最高，其次是茎部；植物激素GA、MeJA、SA处理及条锈菌接种和干旱、低温处理，均能诱导该基因的表达量迅速增加。【结论】成功克隆了小麦TaAly1基因的CDs区，该基因可能通过依赖GA、MeJA和SA的信号通路参与小麦与条锈病的互作。 |
| 关键词: 小麦 蒜氨酸酶 RT-PCR 克隆 |
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| 基金项目:国家自然科学基金重点项目（30930064）；现代农业产业技术体系建设专项（CARS-3-1-11）；高等学校学科创新引智计划项目（B07049） |
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| Cloning and expression analysis of an alliinase gene TaAly1 from wheat |
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| Abstract: |
| 【Objective】The CDs region of wheat alliinase gene was cloned and the sequence characterization and quantitative analysis of expression profiles were performed.【Method】The alliinase gene was isolated from wheat leaves through in silico cloning and reverse transcription PCR (RT-PCR) approaches.The sequence was characterized by software on internet.The expression profiles of the gene under different conditions were analyzed through quantitative real-time PCR (qRT-PCR).【Result】An alliinase gene named TaAly1 was cloned and the open reading frame of TaAly1 was 1 023 bp in length and predicted to encode 340 amino acids protein.The amino acid sequence of TaAly1 shared an identity of 70% with the homologs in rice (Oryza sativa) and maize (Zea mays).The genome sequence contained 3 intron.The TaAly1 gene was highly expressed in leaves and stems than in roots.The expression of TaAly1 was up-regulated after Pst infection and abiotic stress,including GA,MeJA,SA,drought and low temperature treatment.【Conclusion】The CDs region of wheat TaAly1 gene was cloned and the gene may involve in the interaction between Pst and wheat through GA,MeJA and SA dependent signal pathway. |
| Key words: wheat alliinase RT-PCR cloning |
