| 新耐药基因HA117多克隆抗体的制备与鉴定 |
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| 引用本文： | 陈建飞,金先庆,丁雄辉,孙艳辉,赵利华,郭振华,王士奇. 新耐药基因HA117多克隆抗体的制备与鉴定[J]. 第三军医大学学报, 2011, 33(11): 1128-1131 |
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| 作者姓名： | 陈建飞 金先庆 丁雄辉 孙艳辉 赵利华 郭振华 王士奇 |
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| 作者单位： | 重庆医科大学附属儿童医院儿童发育疾病研究省部共建教育部重点实验室,重庆,400014 |
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| 基金项目： | 国家自然科学基金面上项目 |
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| 摘 要： | 目的制备HA117多克隆抗体。方法以HA117表达质粒pAdTrack-HA117为模板,PCR扩增HA117基因,扩增产物转入BL21(DE3)感受态细胞,进行HA117蛋白诱导表达,采用SDS-PAGE检测表达产物。纯化HA117蛋白后免疫新西兰大白兔,制备多克隆抗体。通过间接ELISA检测其效价。将转染携带HA117重组腺病毒的HEK293细胞为实验组,未转染的HEK293细胞为对照组,用细胞免疫荧光和Western blot方法检测其过表达的HA117蛋白,验证制备的HA117多克隆抗体的特异性。结果成功获得HA117多克隆抗体,间接ELISA法检测显示抗HA117血清效价为1∶729 000。细胞免疫荧光和Western blot检测示制备的HA117多克隆抗体可以特异性识别HA117抗原蛋白。结论获得了效价和特异性较高的HA117多克隆抗体。
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| 关 键 词： | HA117 多克隆抗体 抗体制备 鉴定 |
Preparation and identification of polyclonal antibodies of novel multidrug resistance gene HA117 |
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| Chen Jianfei,Jin Xianqing,Ding Xionghui,Sun Yanhui,Zhao Lihua,Guo Zhenhua,Wang Shiqi. Preparation and identification of polyclonal antibodies of novel multidrug resistance gene HA117[J]. Acta Academiae Medicinae Militaris Tertiae, 2011, 33(11): 1128-1131 |
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| Authors: | Chen Jianfei Jin Xianqing Ding Xionghui Sun Yanhui Zhao Lihua Guo Zhenhua Wang Shiqi |
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| Affiliation: | Chen Jianfei,Jin Xianqing,Ding Xionghui,Sun Yanhui,Zhao Lihua,Guo Zhenhua,Wang Shiqi(Key Laboratory of Developmental Diseases in Childhood Cofounded by Chongqing and Ministry of Education,Children's Hospital,Chongqing Medical University,Chongqing,400014,China) |
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| Abstract: | Objective To prepare polyclonal antibodies of a novel multidrug resistance gene,HA117.Methods pAdTrack-HA117 plasmid was used as the template to amplify the total cDNA of HA117 by using PCR.The products of PCR were transferred into BL21(DE3) competent cells to induce the protein expression of HA117 and identified by SDS-PAGE.New Zealand rabbits were immunized with HA117 protein after been purified in order to get the polyclonal antibodies of HA117.Indirect ELISA was used to detect the titer of the polyclona... |
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| Keywords: | HA117 |
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