AAV-ITR基因表达微载体在小鼠体内表达

doi:10.13560/j.cnki.biotech.bull.1985.2016.05.033

生物技术通报 ›› 2016, Vol. 32 ›› Issue (5): 246-254.doi: 10.13560/j.cnki.biotech.bull.1985.2016.05.033

AAV-ITR基因表达微载体在小鼠体内表达

李泰明¹, 许麒麟¹, 潘俊杰², 刘晓玫², 张春²

1. 中国药科大学生命科学与技术学院，南京 210009;
2. 中国科学院苏州生物医学工程技术研究所中国科学院生物医学检验技术重点实验室，苏州 215163

收稿日期:2015-08-03 出版日期:2016-05-25 发布日期:2016-05-27
作者简介:李泰明，女，博士，研究方向：对糖尿病有治疗作用的功能肽研究；E-mail：taimingli@163.com；张春，男，博士，研究方向：基因治疗；
基金资助:
国家自然科学基金项目（81371670），江苏省自然科学基金项目（BK20140381）

Gene Expression of AAV-ITR Mini Vector in Mice vivo

LI Tai-ming¹,XU Qi-lin¹,PAN Jun-jie²,LIU Xiao-mei²,ZHANG Chun²

1. School of Life Science and Technology，China Pharmaceutical University，Nanjing 210009;
2. Key Laboratory of Biomedical Inspection Technology，Suzhou Institute of Biomedical Engineering and Technology，Chinese Academy of Sciences，Suzhou 215163

Received:2015-08-03 Published:2016-05-25 Online:2016-05-27

摘要/Abstract

摘要： 旨在比较AAV-ITR基因表达微载体及其单体在小鼠不同部位的表达差异，将相同拷贝数的AAV-ITR基因表达微载体和pUC57-minivector-EGFP分别转入小鼠脑组织，另外将相同拷贝数的AAV-ITR基因表达微载体单体，AAV-ITR基因表达微载体和pUC57-minivector-EGFP转入小鼠骨骼肌中，比较不同时期组织中的残余分子数量。荧光定量PCR、显微荧光观察以及荧光平均光密度和荧光灰度值分析表明，相同拷贝数的AAV-ITR基因表达微载体单体比AAV-ITR基因表达微载体和pUC57-minivector-EGFP具有更高的转染效率，其稳定性也优于AAV-ITR基因表达微载体。结果显示，AAV-ITR基因表达微载体单体在动物体内具有高效表达安全稳定的特点，有望成为基因治疗中一种安全可靠，稳定高效的新型载体。

关键词: 基因治疗, 基因表达微载体, 腺相关病毒（AAV）, RT-PCR, 组织切片

Abstract: In order to compare the expression difference between AAV-ITR expression mini vector and its monomers in different parts of mice，the same copy number of AAV-ITR mini vector and pUC57-minivector-EGFP were separately transferred into the brain of mice. In addition，the same copy number of monomers of AAV-ITR mini vector and pUC57-minivector-EGFP were transferred into mouse skeletal muscle，and the number of residual molecules were compared in the different tissues at different periods. Analysis by florescence quantitative PCR，microscope observation，average optical density，and gray value showed that monomers of AAV-ITR mini vector in the same copy number had higher transfection efficiency and more stable than AAV-ITR mini vector and pUC57-minivector-EGFP. The comprehensive results suggest that the monomers of AAV-ITR mini vector present the feature of efficient，secure and stable expression，which may be expected to become a novel，secure，reliable，stable，and efficient vector in gene therapy.

Key words: gene therapy, gene expression mini vector, adeno-associated virus（AAV）, RT-PCR, skeletal muscle section

李泰明, 许麒麟, 潘俊杰, 刘晓玫, 张春. AAV-ITR基因表达微载体在小鼠体内表达[J]. 生物技术通报, 2016, 32(5): 246-254.

LI Tai-ming,XU Qi-lin,PAN Jun-jie,LIU Xiao-mei,ZHANG Chun. Gene Expression of AAV-ITR Mini Vector in Mice vivo[J]. Biotechnology Bulletin, 2016, 32(5): 246-254.

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AAV-ITR基因表达微载体在小鼠体内表达

Gene Expression of AAV-ITR Mini Vector in Mice vivo

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