禾谷缢管蚜RpGST1基因的克隆、序列分析及原核表达  

Cloning,sequence analysis and prokaryotic expression of glutathione S-transferase gene from Rhopalosiphum padi( Hemiptera: Aphididae)

作　　者：罗晨[1] 于永昂[2] 左亚运 张改生[2] 赵惠燕[1] 胡祖庆[1] 

机构地区：[1]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 [2]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

出　　处：《植物保护学报》2014年第6期665-672,共8页Journal of Plant Protection

基　　金：西北农林科技大学基本科研业务费专项(QN2013011);西北农林科技大学博士科研启动基金(2013BSJJ114);陕西省科学技术研究发展计划(2013K02-12)

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c DNA序列,命名为Rp GST1(Gen Bank登录号KP192850),并构建原核表达载体p ET32-Rp GST1,对Rp GST1基因进行原核表达、SDS-PAGE和Western blotting检测。结果显示,禾谷缢管蚜Rp GST1基因的编码区长651 bp,编码216个氨基酸,分子量约为24.06 k D,理论等电点为6.20;Rp GST1基因在大肠杆菌中成功表达出一个分子量约为45 k D的融合蛋白,与预测的融合蛋白分子量大小一致。通过克隆Rp GST1基因的c DNA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。In order to clarify the resistance mechanism of Rhopalosiphum padi to pesticides at molecular level,the c DNA sequences of GST genes of R. padi were cloned using RT-PCR and designated as Rp GST1( Gen Bank No. KP192850). The prokaryotic expression of Rp GST1 gene was done after construction of its prokaryotic expression vector p ET32-Rp GST1, SDS-PAGE and Western blotting analysis. The results showed that Rp GST1 gene included an open reading frame of 651 bp encoding 216 amino acids with the predicted molecular mass of 24. 06 k D,and the theoretical isoelectric point was6. 20. The molecular weight of the recombinant Rp GST1 is 45 k D,which is consistent with the predicted result. The c DNA sequences of GST gene from R. padi were successfully cloned and comparatively analyzed. The fusion protein was expressed in vitro by p ET system.

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 

分 类 号：S433.89[农业科学—农业昆虫与害虫防治]