| 卵巢癌相关基因HE4在杆状病毒表达系统的表达与鉴定 |
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| 引用本文： | 陈瑶,陈洁晶,牟霞,李明,吴英松.卵巢癌相关基因HE4在杆状病毒表达系统的表达与鉴定[J].中国妇幼保健,2010,25(2). |
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| 作者姓名： | 陈瑶 陈洁晶 牟霞 李明 吴英松 |
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| 作者单位： | 南方医科大学生物技术学院,广东,广州,510515 |
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| 基金项目： | 广东省卫生厅医学科研基金(A2006375) |
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| 摘 要： | 目的:使卵巢癌相关基因HE4在杆状病毒表达系统中获得表达。方法:将卵巢癌相关基因HE4克隆到pMagic-his-HE4中,通过转化E.coliDH10B筛选克隆,阳性克隆通过接合转移的方式与杆状病毒框架质粒体外重组,构建Bac-phes-HE4杆状病毒表达载体,后者经Lipofectamine2000介导转染Sf9细胞,获取重组病毒,扩增病毒并感染Sf9细胞进行表达,SDS-PAGE、Western blot分析鉴定表达蛋白。结果:经HE4抗体、6×His单抗鉴定,成功通过杆状病毒表达系统获得重组卵巢癌相关基因HE4。结论:卵巢癌相关基因HE4通过接合转移的方法在杆状病毒表达系统中成功表达。
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| 关 键 词： | 克隆细胞 Sf9细胞 杆状病毒科 HE4基因 病毒 基因表达 |
Expression and identification of HE4 from human ovarian cancer in baculovirus expression system |
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| CHEN Yao,CHEN Jie-Jing,MU Xia,et al..Expression and identification of HE4 from human ovarian cancer in baculovirus expression system[J].Maternal and Child Health Care of China,2010,25(2). |
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| Authors: | CHEN Yao CHEN Jie-Jing MU Xia |
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| Institution: | CHEN Yao,CHEN Jie-Jing,MU Xia,et al.School of Biotechnology,Southern Medical University,Guangzhou 510515,Guangdong,China |
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| Abstract: | Objective:To investigate the expression of he4 from human ovarian cancer in baculovirus expression system.Methods:The he4 gene was cloned into the plasmid pMagic-his-HE4.The plasmid pMagic-his-HE4 was transformed into DH10B E.coli to selective positive clone.The HE4 gene was transformed from donor plasmid pMagic-his-HE4 to the recipient plasmid Bac-phes and construct the recombinant vector Bac-phes-HE4.Bac-phes-HE4 transformed into Sf9 cells via Lipofectamine 2000 Reagent to generate the recombinant baculov... |
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| Keywords: | Clone cells Sf9 cells Baculovirus HE4 genes Viral Gene expression |
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