[1]郭玉霞,金先庆,罗庆,等.重组腺病毒AdeasyHA117载体的构建及其在K562细胞的表达*[J].郑州大学学报(医学版),2009,(03):513-515.
 GUO Yuxia,JIN Xianqing,LUO Qing,et al.Obtaining of high titer recombined adenovirus carring HA117 gene and its expression in K562 cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(03):513-515.
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重组腺病毒AdeasyHA117载体的构建及其在K562细胞的表达*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年03期
页码:
513-515
栏目:
论著
出版日期:
2009-05-01

文章信息/Info

Title:
Obtaining of high titer recombined adenovirus carring HA117 gene and its expression in K562 cells
作者:
郭玉霞金先庆罗庆郑改焕于洁刘筱梅温贤浩徐酉华#
重庆医科大学附属儿童医院血液科重庆400014
Author(s):
GUO YuxiaJIN XianqingLUO QingZHENG GaihuanYU JieLIU XiaomeiWEN XianhaoXU Youhua
Department of Hematology, Children’s Hospital of Chongqing Medical University, Chongqing 400014
关键词:
耐药相关基因HA117腺病毒K562细胞
Keywords:
resistance gene HA117adenovirusK562 cells
分类号:
R733.7
摘要:
构建携带耐药相关基因HA117的高滴度重组腺病毒,转染K562细胞,检测其在K562细胞中的表达情况。方法:用基因工程技术将ATRA相关耐药新基因HA117的cDNA亚克隆至穿梭质粒pAdTrackCMV上,然后在细菌内与pAdeasy同源重组,后将重组腺病毒AdeasyHA117经脂质体转染包装细胞HEK293,并收获原代的重组腺病毒Ad5easyHA117,经乒乓交互感染的方法扩增原代腺病毒,应用RTPCR、荧光显微镜及流式细胞仪鉴定和检测重组腺病毒感染K562细胞后HA117的表达情况。结果:酶切鉴定及PCR结果证明ATRA相关耐药新基因HA117重组腺病毒载体构建成功,携带新基因HA117重组腺病毒载体的效价达8.3×1011pfu/mL。RTPCR检测到转染后K562细胞内HA117的表达。结论:成功获得携带新基因HA117的重组腺病毒,并能使其在K562细胞中高表达。
Abstract:
To obtain the high titer recombinant adenovirus expressing human HA117 gene.Methods:The cDNA of HA117 gene was subcloned into the shuttle plasmid pAdTrackCMV and then clone the homologous recombinant adenovims genomie plasmid pAdeasy in bacteria.We tranfected the DNA of identified recombinant plasmid into 293 ceils via liposome,and then did the package and amplification of adenovims, determined by fluorescence microscope.

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目30471985#通讯作者,女,1945年生,教授,研究方向:肿瘤的基因治疗,Email:etzhl@163.com
更新日期/Last Update: 2010-05-28