|本期目录/Table of Contents|

[1]赵景川,严巧灵,钱其军. 靶向肝癌细胞的嵌合AAV建库及筛选[J].浙江理工大学学报,2012,29(06):874-878.
　ZHAO Jing chuan,YAN Qiao ling,QIAN Qi jun. The Libraries Building and Screening of Chimeric AAVTargeting Liver Cancer Cells[J].Journal of Zhejiang Sci-Tech University,2012,29(06):874-878.
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 靶向肝癌细胞的嵌合AAV建库及筛选()
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浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第29卷
期数:
2012年06期
页码:
874-878
栏目:
(自科)生物与生命科学
出版日期:
2012-12-28

文章信息/Info

Title:
 The Libraries Building and Screening of Chimeric AAVTargeting Liver Cancer Cells
文章编号:
16733851 (2012) 06087405
作者:
 赵景川1;  严巧灵2;  钱其军1; 2
 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医院 病毒基因治疗实验室, 上海 200438
Author(s):
 ZHAO Jingchuan1;  YAN Qiaoling2;  QIAN Qijun1; 2
 1. Xinyuan Institute of Medicine and Biotechnology, School of Life Sciences, Zhejiang SciTechUniversity, Hangzhou 310018, China; 2. Laboratory of Virus and Gene Therapy, EasternHepatobiliary Surgical Hospital, Second Military Medical University, Shanghai 200438, China
关键词:
 ; Shuffling;  嵌合AAV;  靶向;  肝癌细胞
分类号:
Q782
文献标志码:
A
摘要:
    采用Shuffling的方法,将AAV19 Cap基因序列进行改组,筛选出高效靶向肝癌细胞的嵌合AAV载体。以pAAVMCS载体为骨架,构建出含AAV2 ITR和Rep2序列的pAdB载体,在此基础上,构建含有嵌合Cap序列的pARC载体库,用于转染293细胞,并加入5型腺病毒（Ad5）,包装出嵌合AAV病毒。然后将包装出的嵌合AAV病毒与Ad5共感染BEL7402肝癌细胞,经过三轮筛选后,初步筛选出可以靶向肝癌细胞的嵌合AAV。最后将筛选出的AAV Rep2嵌合Cap基因的融合片段连接到pMD18T载体上,构建出pARCT载体,与AAVEGFP质粒共转染293细胞,并加入Ad5,包装出病毒基因组含EGFP基因、病毒外壳为嵌合Cap的AAV病毒（cAAVEGFP）,并用其感染BEL7402细胞,观察绿色荧光的表达。最后观察到绿色荧光表达,证明筛选出了靶向BEL7402肝癌细胞的嵌合AAV。

参考文献/References:

 ［1］ Sharma A, Ghosh A, Hansen E T, et al. Transduction efficiency of AAV 2/6, 2/8 and 2/9 vectors for delivering genes in human corneal fibroblasts［J］. Brain Research Bulletin, 2010, 81(23): 273278.
［2］ Foust K D, Nurre E, Montgomery C L, et al. Intravascular AAV9 preferentially targets neonatal neurons and adult astrocytes［J］. Nat Biotechnol, 2009, 27(1): 5965.〖JP〗〖LL〗
［3］ Ku C A, Chiodo V A, Boye S L, et al. Gene therapy using selfcomplementary Y733F capsid mutant AAV2/8 restores vision in a model of early onset Leber congenital amaurosis［J］. Hum Mol Genet, 2011, 20(23): 45694581
［4］ Li H, Malani N, Hamilton S R, et al. Assessing the potential for AAV vector genotoxicity in a murine model［J］, Blood, 2011, 117(12): 33113319.
［5］ Aartsen W M, van Cleef K W, Pellissier L P, et al. GFAPdriven GFP expression in activated mouse Müller glial cells aligning retinal blood vessels following intravitreal injection of AAV2/6 vectors［J］. PLoS ONE, 2010, 5(8): e12387.
［6］ Stemmer W P. DNA shuffling by random fragmentation and reassembly: in vitro recombination for molecular evolution［J］. Proc Natl Acad Sci U S A, 1994, 91(22): 1074710751.
［7］ Stemmer W P. Rapid evolution of a protein in vitro by DNA shuffling［J］. Nature, 1994, 370(6488): 389391.
［8］ Rosic N N, Huang W, Johnston W A, et al. Extending the diversity of cytochrome P450 enzymes by DNA family shuffling［J］. Gene, 2007, 395(12): 4048.
［9］ Yang L, Jiang J, Drouin L M, et al. A myocardium tropic adenoassociated virus (AAV) evolved by DNA shuffling and in vivo selection［J］. Proc Natl Acad Sci U S A， 2009, 106(10): 39463951.
［10］ 单世平, 夏立秋, 丁学知, 等. DNA改组的最新动态及应用前景［J］. 中国生物工程杂志， 2009, 29(10): 9297.

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备注/Memo

备注/Memo:
 收稿日期： 2012-02-15
基金项目： 赵景川（1983-）,男,山东临沂人,硕士研究生,主要从事肿瘤的靶向基因病毒治疗方面的研究。
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