| hGITRL_(aa52-177)蛋白在杆状病毒系统中的表达 |
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| 引用本文： | 唐莉,王胜军,毛朝明,陈君,胡正军,鲍俊峰,仝佳,邵启祥,许化溪.hGITRL_(aa52-177)蛋白在杆状病毒系统中的表达[J].免疫学杂志,2009,25(2). |
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| 作者姓名： | 唐莉 王胜军 毛朝明 陈君 胡正军 鲍俊峰 仝佳 邵启祥 许化溪 |
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| 作者单位： | 江苏大学医学技术学院免疫学和免疫检验学系,江苏,镇江,212013
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| 基金项目： | 江苏省自然科学基金，江苏大学拔尖人才工程和高级人才基金，国家自然科学基金 |
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| 摘 要： | 目的 利用杆状病毒系统表达hGITRLaa52-177蛋白.方法 PCR扩增获得hGITRL胞外段(hGITRLaa52-177)基因,克隆入pFastBacHTa转移载体,在E.coli DH10Bac内通过转座子Tn7的介导,得到重组杆粒,转染Tn细胞表达目的蛋白.通过SDS-PAGE电泳及Western blotting鉴定目的蛋白的表达.结果 重组载体 pFastBacHTa-hGITRLaa52-177以及重组杆粒Bacmid-hGITRLaa52-177构建正确,在Tn细胞内成功表达6×His-hGITRLaa52-177重组蛋白,SDS-PAGE电泳及Western blotting显示重组hGITRLaa52-177蛋白相对分子质量约为Mr 18 000.结论 杆状病毒-昆虫表达系统获得 hGITRLaa52-177蛋白,为进一步从蛋白质水平研究hGITR/hGITRL对机体免疫调节的影响奠定了基础.
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| 关 键 词： | 杆状病毒 真核表达 |
Human GITRL_(aa52-177) protein expressed in Bac-to-Bac baculovirus expression system |
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| TANG Li,WANG Shengjun,MAO Chaoming,CHEN Jun,HU Zhengjun,BAO Junfeng,TONG Jia,SHAO Qixiang,XU Huaxi.Human GITRL_(aa52-177) protein expressed in Bac-to-Bac baculovirus expression system[J].Immunological Journal,2009,25(2). |
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| Authors: | TANG Li WANG Shengjun MAO Chaoming CHEN Jun HU Zhengjun BAO Junfeng TONG Jia SHAO Qixiang XU Huaxi |
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| Institution: | TANG Li,WANG Shengjun,MAO Chaoming,CHEN Jun,HU Zhengjun,BAO Junfeng,TONG Jia,SHAO Qixiang,XU Huaxi Department of Immunology,School of Medical Technology,Jiangsu University,Zhengjiang 212013,China |
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| Abstract: | Objective To produce the extracellular region of hGITRL in eukaryotic expression system.Methods Bac-to-Bac baculovirus expression system was employed in the present study.The hGITRL extracellular region gene was amplified by PCR from the full-length pMD18-hGITRL-T vector and subcloned into the pFastBacHTa plasmid,then the latter was transformed into E.coli DH10Bac competent cells to construct a recombinant baculovirus and to infect Tn cells.The interest protein was identified by SDS-PAGE,Western blot and in... |
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| Keywords: | GITRL |
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