| 柑橘溃疡病菌抗铜相关基因copA和 copB的克隆及原核表达 |
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| 引用本文： | 王君,叶刚,马燕侠,王国平,洪霓. 柑橘溃疡病菌抗铜相关基因copA和 copB的克隆及原核表达[J]. 植物病理学报, 2011, 41(3): 247-252 |
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| 作者姓名： | 王君 叶刚 马燕侠 王国平 洪霓 |
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| 作者单位： | 1华中农业大学植物科技学院 湖北省作物病害监测与安全控制重点实验室，武汉 430070； 2华中农业大学 农业微生物学国家重点实验室，武汉 430070 |
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| 基金项目： | 国家科技支撑计划项目，农业行业专项计划 |
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| 摘 要： | 以柑橘溃疡病菌DNA为模板，对抗铜相关基因copA和copB进行了PCR扩增和克隆，获得了大小分别为1 782 bp 和1 095 bp的目标片段。构建了这2个基因的原核表达载体 (pET-copA和pET-copB)，并在大肠杆菌 [Escherichia coli BL21(DE3)] 中成功诱导表达。利用原核表达的融合蛋白免疫大耳白兔，制备了抗copA和copB原核表达蛋白的多克隆抗体。用间接ELISA法测定了所制备的多克隆抗体的效价均为1∶6 400。用所制备的抗体分别对copA和copB的原核表达产物进行Western blot分析的结果显示，在相应位置产生了较强的免疫反应条带，表明所制备抗体能特异性地与相应的抗原发生免疫反应。
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| 关 键 词： | 抗铜基因 PCR 原核表达 Western blot 柑橘溃疡病菌 |
| 收稿时间： | 2010-07-12; |
Cloning and prokaryotic expression of the copper-resistance related genes copA and copB of Xanthomonas citri subsp.citri |
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| WANG Jun,YE Gang,MA Yan-xia,WANG Guo-ping,HONG Ni. Cloning and prokaryotic expression of the copper-resistance related genes copA and copB of Xanthomonas citri subsp.citri[J]. Acta Phytopathologica Sinica, 2011, 41(3): 247-252 |
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| Authors: | WANG Jun YE Gang MA Yan-xia WANG Guo-ping HONG Ni |
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| Affiliation: | WANG Jun1,YE Gang1,MA Yan-xia1,WANG Guo-ping1,2,HONG Ni1,2 (1College of Plant Science and Technology,Huazhong Agricultural University,Key Lab of Crop Disease Monitoring and Safety Control in Hubei,Wuhan 430070,China,2Huazhong Agricultural University,National Key Lab of Agromicrobiology,China) |
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| Abstract: | Copper-resistance related genes copA and copB were amplified by using the DNA as template extracted from Xanthomonas citri subsp.citri.The target products were cloned,and sequencing results shows that sizes of copA and copB with 1 782 bp and 1 095 bp.The prokaryotic expression vectors pET-copA and pET-copB for these two genes were constructed.Both copA and copB were effectively expressed as fusion proteins in transformed Escherichia coli strain BL21(DE3) under the induction of IPTG.The expressed proteins we... |
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| Keywords: | copper-resistance related gene PCR prokaryotic expression Western blot Xanthomonas citri subsp.citri |
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