| 禾谷缢管蚜RpGST1基因的克隆、序列分析及原核表达 |
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| 引用本文： | 罗晨,于永昂,左亚运,张改生,赵惠燕,胡祖庆.禾谷缢管蚜RpGST1基因的克隆、序列分析及原核表达[J].植物保护学报,2014,41(6):665-672. |
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| 作者姓名： | 罗晨 于永昂 左亚运 张改生 赵惠燕 胡祖庆 |
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| 作者单位： | 西北农林科技大学植物保护学院, 旱区作物逆境生物学国家重点实验室, 陕西 杨凌 712100;西北农林 科技大学, 国家杨凌农业生物技术育种中心, 国家小麦改良中心杨凌分中心, 小麦育种教育部工程研究中心, 陕西省作物杂种优势研究与利用重点实验室, 杨凌 712100;西北农林科技大学植物保护学院, 旱区作物逆境生物学国家重点实验室, 陕西 杨凌 712100;西北农林 科技大学, 国家杨凌农业生物技术育种中心, 国家小麦改良中心杨凌分中心, 小麦育种教育部工程研究中心, 陕西省作物杂种优势研究与利用重点实验室, 杨凌 712100;西北农林科技大学植物保护学院, 旱区作物逆境生物学国家重点实验室, 陕西 杨凌 712100;西北农林科技大学植物保护学院, 旱区作物逆境生物学国家重点实验室, 陕西 杨凌 712100 |
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| 基金项目： | 国家自然科学基金(31272036,31471766),教育部高等学校博士学科点专项科研基金(20110204110001) |
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| 摘 要： | 为揭示禾谷缢管蚜耐药性的分子机理,采用RT-PCR方法克隆了禾谷缢管蚜谷胱甘肽-S-转移酶(glutathione S-transferases,GSTs)的cDNA序列,命名为RpGST1(GenBank登录号KP192850),并构建原核表达载体pET32-RpGST1,对RpGST1基因进行原核表达、SDS-PAGE和Western blotting检测。结果显示,禾谷缢管蚜RpGST1基因的编码区长651 bp,编码216个氨基酸,分子量约为24.06 kD,理论等电点为6.20;RpGST1基因在大肠杆菌中成功表达出一个分子量约为45 kD的融合蛋白,与预测的融合蛋白分子量大小一致。通过克隆RpGST1基因的cDNA序列并进行序列比对分析,表明构建了GST基因的原核表达载体并成功表达。
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| 关 键 词： | 禾谷缢管蚜 谷胱甘肽-S-转移酶 基因克隆 序列分析 原核表达 |
| 收稿时间： | 2014/11/15 0:00:00 |
Cloning, sequence analysis and prokaryotic expression of glutathione S-transferase gene from Rhopalosiphum padi (Hemiptera: Aphididae) |
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| Luo Chen,Yu Yong''ang,Zuo Yayun,Zhang Gaisheng,Zhao Huiyan and Hu Zuqing.Cloning, sequence analysis and prokaryotic expression of glutathione S-transferase gene from Rhopalosiphum padi (Hemiptera: Aphididae)[J].Acta Phytophylacica Sinica,2014,41(6):665-672. |
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| Authors: | Luo Chen Yu Yong'ang Zuo Yayun Zhang Gaisheng Zhao Huiyan and Hu Zuqing |
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| Institution: | State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A & F University, Yangling 712100, Shaanxi Province, China;Northwest A & F University; National Yangling Agricultural Biotechnology & Breeding Center; Yangling Branch of State Wheat Improvement Center; Wheat Breeding Engineering Research Center, Ministry of Education; Key Laboratory of Crop Heterosis of Shaanxi Province, Yangling 712100, Shaanxi Province, China;State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A & F University, Yangling 712100, Shaanxi Province, China;Northwest A & F University; National Yangling Agricultural Biotechnology & Breeding Center; Yangling Branch of State Wheat Improvement Center; Wheat Breeding Engineering Research Center, Ministry of Education; Key Laboratory of Crop Heterosis of Shaanxi Province, Yangling 712100, Shaanxi Province, China;State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A & F University, Yangling 712100, Shaanxi Province, China;State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A & F University, Yangling 712100, Shaanxi Province, China |
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| Abstract: | In order to clarify the resistance mechanism of Rhopalosiphum padi to pesticides at molecular level, the cDNA sequences of GST genes of R. padi were cloned using RT-PCR and designated as RpGST1 (GenBank No. KP192850). The prokaryotic expression of RpGST1 gene was done after construction of its prokaryotic expression vector pET32-RpGST1, SDS-PAGE and Western blotting analysis. The results showed that RpGST1 gene included an open reading frame of 651 bp encoding 216 amino acids with the predicted molecular mass of 24.06 kD, and the theoretical isoelectric point was 6.20. The molecular weight of the recombinant RpGST1 is 45kD, which is consistent with the predicted result. The cDNA sequences of GST gene from R.padi were successfully cloned and comparatively analyzed. The fusion protein was expressed in vitro by pET system. |
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| Keywords: | Rhopalosiphum padi glutathione S-transferases gene cloning sequence analysis prokaryotic expression |
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