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[1]贾玉,李林芳,钱其军. 携带HerceptinMICB融合蛋白基因的腺病毒载体的构建[J].浙江理工大学学报,2011,28(01):111-115.
　JIA Yu,LI Lin fang,QIAN Qi jun. Construction and Identification of a Recombinant Adenovirus Vector Expressing HerceptinMICB Fusion Protein[J].Journal of Zhejiang Sci-Tech University,2011,28(01):111-115.
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 携带HerceptinMICB融合蛋白基因的腺病毒载体的构建()
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浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第28卷
期数:
2011年01期
页码:
111-115
栏目:
(自科)生物与生命科学
出版日期:
2011-02-28

文章信息/Info

Title:
 Construction and Identification of a Recombinant Adenovirus Vector Expressing HerceptinMICB Fusion Protein
文章编号:
16733851 (2011) 01011105
作者:
 贾玉1;  李林芳2;  钱其军1; 2
 1. 浙江理工大学新元医学与生物技术研究所， 杭州 310018； 2. 第二军医大学东方肝胆外科医院， 上海 200438
Author(s):
 JIA Yu1;  LI Linfang2;  QIAN Qijun1; 2
 1. Xinyuan Institute of Medicine and Biotechnology, Zhejiang SciTech University, Hangzhou 310018, China; 2. Eastern Hepatobiliary Surgical Hospital, The Second Militaty Medical University, Shanghai 200438, China
关键词:
 ; 肿瘤;  抗体治疗;  Herceptin;  NKG2D;  融合蛋白
分类号:
Q782
文献标志码:
A
摘要:
    为了构建携带HerceptinMICB融合蛋白基因的腺病毒载体，筛选出能表达该融合蛋白的重组腺病毒，检测其表达融合蛋白的情况及抗肿瘤特征。首先，通过PCR将NKG2D的配体MICB的基因片段插入含有全长抗体Herceptin基因的5型腺病毒穿梭载体pDC339Her中，获得载体pDC339HerMICB与腺病毒骨架载体pBHGloxΔE1,3Cr重组，然后在293细胞内进行病毒包装得到非增殖型腺病毒AdHerMICB，纯化后测病毒滴度。双抗体夹心ELISA和Western blot检测融合蛋白的表达情况；间接免疫荧光实验(IFA)检测融合蛋白的特异性。酶切后DNA电泳鉴定证实载体构建成功，重组病毒经PCR鉴定携带融合蛋白基因。融合蛋白的表达量为（623.25±38.62）ng/mL；Westhern blot显示：该融合蛋白与商品化的Herceptin抗体轻链重链比较，大小一致，浓度匹配；间接免疫荧光检测（IFA）显示了融合蛋白与HER2过表达的卵巢癌细胞株SKOV3结合的特异性。结果表明：腺病毒AdHerMICB携带HerceptinNKG2D配体融合蛋白基因能在体外正常表达且与商品化的Herceptin生物学特性相似，为融合蛋白的抗肿瘤治疗奠定基础。

参考文献/References:

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备注/Memo

备注/Memo:
 收稿日期： 2010-01-13
基金项目： 国家新药创制重大专项课题（2009ZX09103687）
作者简介： 贾玉（1983-），男，安徽蚌埠人，硕士研究生，主要从事肿瘤靶向基因病毒治疗及抗体治疗方面的研究。
通讯作者： 钱其军，电子邮箱:qianqj@163.com
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