|本期目录/Table of Contents|

[1]刘海燕,钱其军.利用杆状病毒系统获得携带抗体表达基因的双链AAV的研究[J].浙江理工大学学报,2013,30(06):870-876.
　LIU Hai yan,QIAN Qi jun.Study on Production of Doublestrand AAV Carrying AntibodyExpressed Gene Using Baculovirus System[J].Journal of Zhejiang Sci-Tech University,2013,30(06):870-876.
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利用杆状病毒系统获得携带抗体表达基因的双链AAV的研究()
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浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第30卷
期数:
2013年06期
页码:
870-876
栏目:
(自科)生物与生命科学
出版日期:
2013-11-10

文章信息/Info

Title:
Study on Production of Doublestrand AAV Carrying AntibodyExpressed Gene Using Baculovirus System
文章编号:
1673-3851 (2013) 06-0870-07
作者:
刘海燕;  钱其军
1. 浙江理工大学生命科学学院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病毒基因治疗实验室, 上海 200438
Author(s):
LIU Hai yan;  QIAN Qi jun
1. School of Life Science, Zhejiang Sci Tech University, Hangzhou 310018, China;2. Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgical Hospital,Second Military Medical University, Shanghai 200438, China
关键词:
杆状病毒; 双链AAV;  病毒生产;  外源
分类号:
Q782
文献标志码:
A
摘要:
研究使用二杆状病毒昆虫细胞系统获得携带抗体表达基因的8型双链腺相关病毒(scAAV)。首先利用杆状病毒系统获得ssrAAV8 EGFP和scrAAV8 EGFP，实验表明scrAAV8 EGFP比ssrAAV8 EGFP具有更高的转导效率；然后以同样的方法获得scrAAV8 AT7，检测其滴度，并感染HEK293细胞以检测抗体的表达情况。蛋白印迹法检测出阿瓦斯汀(Avastin)重轻链已成功表达，ELISA检测出Avastin在体外的表达量可达770 ng/mL。该研究表明利用杆状病毒系统所获得8型scAAV具有生物学活性并能实现外源基因的体外表达。

参考文献/References:

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［2］ Ferrari F K, Samulski T, Shenk T, et al. Secondstrand synthesis is a ratelimiting step for efficient transduction by recombinant adenoassociated virus vectors［J］. J Virol, 1996, 70(5): 3227-3234.
［3］ Chen H. Intron splicingmediated expression of AAV Rep and Cap genes and production of AAV vectors in insect cells［J］. Mol Ther, 2008, 16(5): 924-930.
［4］ Nathwani A C, Gray J T, Ng C Y, et al. Selfcomplementary adenoassociated virus vectors containing a novel liverspecific human factor IX expression cassette enable highly efficient transduction of murine and nonhuman primate liver［J］. Blood, 2006, 107(7): 2653-2661.
［5］ Gao G P, Lu Y, Sun X, et al. Highlevel transgene expression in nonhuman primate liver with novel adenoassociated virus serotypes containing selfcomplementary genomes［J］. J Virol, 2006, 80(12): 6192-6194.
［6］ McCarty D M. Selfcomplementary AAV vectors advances and applications［J］. Mol Ther, 2008, 16(10): 1648-1656.
［7］ Wang Yu, Gao Hui, Chen Chuan, et al. Promoter activity of different promoters in recombinant baculovirusinfected Sf9 cells［J］. Chin J Biotech, 2008, 24(4): 598-603.
［8］ 丁娜. 利用杆状病毒系统规模化生产rAAV1的研究［D］. 杭州: 浙江理工大学, 2011.
［9］ Smith R H, Levy J R, Kotin R M. A simplified baculovirusAAV expression vector system coupled with onestep affinity purification yields hightiter rAAV stocks from insect cells［J］. Mol Ther. 2009, 17(11): 1888-1896.

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备注/Memo

备注/Memo:
收稿日期: 2013-03-12
基金项目: 国家自然科学基金(8107185)
作者简介: 刘海燕(1985-),女,山东威海人,硕士研究生,主要从事肿瘤的基因病毒治疗研究。
通信作者: 钱其军,Email:qianqi@163.com
更新日期/Last Update: 2013-11-12