| 摘要: |
| 【目的】克隆小麦条锈菌PsSte12基因，明确其序列特征、转录活性及其在小麦条锈菌侵染过程中的相对表达量。【方法】采用RT-PCR方法，从小麦条锈菌中获得PsSte12基因，利用生物信息学分析其序列特征，实时定量PCR技术检测其在小麦条锈菌不同侵染阶段的表达特征；借助酵母单杂交系统解析该基因的转录活性。【结果】克隆到1个小麦条锈菌基因PsSte12，该基因序列含有1个全长2 553 bp的开放阅读框；基因组DNA序列含有4个内含子，分别位于开放阅读框第281，429，1 512，1 584位，长度分别为73，79，66和68 bp。该基因编码蛋白含850个氨基酸,75个正电荷残基和90个负电荷残基。PsSte12与小麦秆锈菌、杨树锈菌、小麦赤霉菌及构巢曲霉菌中同源序列的相似性分别为87.9%，61.53%，24.3%和25.2%；PsSte12在小麦条锈菌侵染后24和36 h大量表达，相对表达量分别为对照的61倍和123倍；PsSte12具有转录活性，其N端为转录激活区。【结论】成功克隆了小麦条锈菌STE类型转录因子PsSte12，证实其具有转录活性，在小麦条锈菌侵染早期诱导表达，推测其参与了小麦条锈菌的侵染过程。 |
| 关键词: 小麦条锈菌 转录因子PsSte12 基因克隆 转录活性 |
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| 基金项目:国家自然科学基金青年科学基金项目（31201463）；西北农林科技大学基本科研业务费专项（2452015017） |
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| Cloning and characterization of PsSte12,a STE transcription factor of Puccinia striiformis |
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HUANG Xueling,FU Jie
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| Abstract: |
| 【Objective】This study cloned the PsSte12gene from Puccinia striiformis (Pst) and analyzed its sequence characteristics,transcription activity and expression profiles during Pst infection.【Method】The PsSte12 gene was obtained by RT-PCR approach and its sequence characteristics were analyzed using related software.The qRT-PCR was used to determine the expression profiles of PsSte12 during Pst infection,and the transcription activity was examined using yeast one-hybrid assay.【Result】The PsSte12 was obtained with an ORF of 2 553 bp and it contained four introns located at positions 281,429,1 512 and 1 584 of the ORF with the lengths of 73,79,66 and 68 bp,respectively.The gene encoded 850 amino acids including 75 positively charged residues and 90 negatively charged residues.The amino acid sequence shared an identity of 87.9%,61.53%,24.3% and 25.2% with Puccinia f.sp.graminis,Melampsora larici-populina,Fusarium graminearum and Aspergillus nidulans,respectively.PsSte12 was up-regulated post infection and the expressions at 24 and 36 hours were 61 and 123 times of the control.PsSte12 exhibited the transcription activity,and the N terminus was responsible for it.【Conclusion】The Pst STE transcription factor PsSte12 was cloned and identified with transcription activity.It was induced during infection,suggesting that PsSte12may involve in Pst infection. |
| Key words: Puccinia striiformis transcription factor gene clone transcription activation |
