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禾谷缢管蚜RpGST1基因的克隆、序列分析及原核表达

, PP. 665-672

Keywords: 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

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Abstract:

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(glutathioneS-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Westernblotting检测。结果显示,禾谷缢管蚜RpGST1基因的编码区长651bp,编码216个氨基酸,分子量约为24.06kD,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45kD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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