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转LdOA1基因果蝇品系GSTs基因表达及对溴氰菊酯胁迫响应

孙丽丽 刘鹏 王志英 梁斌 曹传旺

孙丽丽, 刘鹏, 王志英, 梁斌, 曹传旺. 转LdOA1基因果蝇品系GSTs基因表达及对溴氰菊酯胁迫响应[J]. 北京林业大学学报, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418
引用本文: 孙丽丽, 刘鹏, 王志英, 梁斌, 曹传旺. 转LdOA1基因果蝇品系GSTs基因表达及对溴氰菊酯胁迫响应[J]. 北京林业大学学报, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418
SUN Li-li, LIU Peng, WANG Zhi-ying, LIANG Bin, CAO Chuan-wang. Glutathione S-transferase gene expression in transformant Drosophila expressing ocular albinism type 1 from gypsy moth and its response to deltamethrin stress[J]. Journal of Beijing Forestry University, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418
Citation: SUN Li-li, LIU Peng, WANG Zhi-ying, LIANG Bin, CAO Chuan-wang. Glutathione S-transferase gene expression in transformant Drosophila expressing ocular albinism type 1 from gypsy moth and its response to deltamethrin stress[J]. Journal of Beijing Forestry University, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418

转LdOA1基因果蝇品系GSTs基因表达及对溴氰菊酯胁迫响应

doi: 10.13332/j.1000-1522.20150418
基金项目: 

国家自然科学基金项目(31570642和31101676)、黑龙江省自然科学基金面上项目(C201409)

详细信息
    作者简介:

    孙丽丽,博士生。主要研究方向:昆虫毒理学。Email: 253035020@qq.com 地址:150040 黑龙江省哈尔滨市和兴路26号东北林业大学林学院。

    通讯作者:

    曹传旺,副教授。主要研究方向:昆虫毒理学。Email: chuanwangcao@126.com 地址:同上。

Glutathione S-transferase gene expression in transformant Drosophila expressing ocular albinism type 1 from gypsy moth and its response to deltamethrin stress

  • 摘要: 舞毒蛾是重要林业食叶害虫,揭示其G蛋白偶联受体介导G蛋白对GSTs的调控,这对于挖掘分子靶标开发新型杀虫剂具有重要意义。本文构建转LdOA1基因果蝇载体,获得表达LdOA1基因果蝇品系,分析了转基因果蝇GSTs基因表达量及对溴氰菊酯胁迫的响应,为明确昆虫OA1基因功能提供理论依据。通过传统的酶切-连接方法构建重组载体pUAST-attB-LdOA1,采用转基因技术构建表达LdOA1基因的纯合果蝇品系,利用PCR技术验证转基因果蝇品系,并运用实时荧光定量RT-PCR技术测定转基因果蝇GSTs Delta家族基因表达量及低浓度溴氰菊酯胁迫对GSTs基因表达量的影响。PCR扩增条带显示转LdOA1基因果蝇品系均能检测到453bp目的基因条带。与非转基因果蝇相比,转LdOA1基因果蝇品系中GSTs Delta家族基因(除GSTd4和GSTd7)均上调表达,为非转基因组1.01~3.27倍。低浓度溴氰菊酯胁迫6h,非转基因果蝇GSTd6和GSTd10基因被显著诱导激活,而其他GSTs基因被显著抑制,抑制率为6.48%~95.84%,胁迫12~72h,GSTs基因(除GSTd1和GSTd10外)均被显著抑制。低浓度溴氰菊酯胁迫转基因果蝇GSTs表达量变化趋势与非转基因果蝇基本一致,但转基因果蝇品系GSTs表达量显著高于非转基因果蝇品系(除12~48h GSTd1和GSTd10),且胁迫6h表达量最大。这些结果表明LdOA1基因可能介导G蛋白信号通路调控下游GSTs Delta亚家族基因表达响应溴氰菊酯胁迫。
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  • 收稿日期:  2015-11-24
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  • 刊出日期:  2016-06-30

转LdOA1基因果蝇品系GSTs基因表达及对溴氰菊酯胁迫响应

doi: 10.13332/j.1000-1522.20150418
    基金项目:

    国家自然科学基金项目(31570642和31101676)、黑龙江省自然科学基金面上项目(C201409)

    作者简介:

    孙丽丽,博士生。主要研究方向:昆虫毒理学。Email: 253035020@qq.com 地址:150040 黑龙江省哈尔滨市和兴路26号东北林业大学林学院。

    通讯作者: 曹传旺,副教授。主要研究方向:昆虫毒理学。Email: chuanwangcao@126.com 地址:同上。

摘要: 舞毒蛾是重要林业食叶害虫,揭示其G蛋白偶联受体介导G蛋白对GSTs的调控,这对于挖掘分子靶标开发新型杀虫剂具有重要意义。本文构建转LdOA1基因果蝇载体,获得表达LdOA1基因果蝇品系,分析了转基因果蝇GSTs基因表达量及对溴氰菊酯胁迫的响应,为明确昆虫OA1基因功能提供理论依据。通过传统的酶切-连接方法构建重组载体pUAST-attB-LdOA1,采用转基因技术构建表达LdOA1基因的纯合果蝇品系,利用PCR技术验证转基因果蝇品系,并运用实时荧光定量RT-PCR技术测定转基因果蝇GSTs Delta家族基因表达量及低浓度溴氰菊酯胁迫对GSTs基因表达量的影响。PCR扩增条带显示转LdOA1基因果蝇品系均能检测到453bp目的基因条带。与非转基因果蝇相比,转LdOA1基因果蝇品系中GSTs Delta家族基因(除GSTd4和GSTd7)均上调表达,为非转基因组1.01~3.27倍。低浓度溴氰菊酯胁迫6h,非转基因果蝇GSTd6和GSTd10基因被显著诱导激活,而其他GSTs基因被显著抑制,抑制率为6.48%~95.84%,胁迫12~72h,GSTs基因(除GSTd1和GSTd10外)均被显著抑制。低浓度溴氰菊酯胁迫转基因果蝇GSTs表达量变化趋势与非转基因果蝇基本一致,但转基因果蝇品系GSTs表达量显著高于非转基因果蝇品系(除12~48h GSTd1和GSTd10),且胁迫6h表达量最大。这些结果表明LdOA1基因可能介导G蛋白信号通路调控下游GSTs Delta亚家族基因表达响应溴氰菊酯胁迫。

English Abstract

孙丽丽, 刘鹏, 王志英, 梁斌, 曹传旺. 转LdOA1基因果蝇品系GSTs基因表达及对溴氰菊酯胁迫响应[J]. 北京林业大学学报, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418
引用本文: 孙丽丽, 刘鹏, 王志英, 梁斌, 曹传旺. 转LdOA1基因果蝇品系GSTs基因表达及对溴氰菊酯胁迫响应[J]. 北京林业大学学报, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418
SUN Li-li, LIU Peng, WANG Zhi-ying, LIANG Bin, CAO Chuan-wang. Glutathione S-transferase gene expression in transformant Drosophila expressing ocular albinism type 1 from gypsy moth and its response to deltamethrin stress[J]. Journal of Beijing Forestry University, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418
Citation: SUN Li-li, LIU Peng, WANG Zhi-ying, LIANG Bin, CAO Chuan-wang. Glutathione S-transferase gene expression in transformant Drosophila expressing ocular albinism type 1 from gypsy moth and its response to deltamethrin stress[J]. Journal of Beijing Forestry University, 2016, 38(6): 72-78. doi: 10.13332/j.1000-1522.20150418
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