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[1]任平,张贺林,牛建国,等.STAT3基因shRNA慢病毒载体的构建与鉴定[J].宁夏医科大学学报,2014,(03):260-262.
 REN Ping,ZHANG Helin,NIU Jianguo,et al.Construction of STAT3 shRNA Lentiviral Vector[J].Ningxia Medical University,2014,(03):260-262.
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STAT3基因shRNA慢病毒载体的构建与鉴定(PDF)
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2014年03期
页码:
260-262
栏目:
论著
出版日期:
2014-03-30

文章信息/Info

Title:
Construction of STAT3 shRNA Lentiviral Vector
作者:
任平1张贺林2牛建国34刘雅琦34顾金海34
1.宁夏吴忠市人民医院神经外科,吴忠 751100; 2.宁夏吴忠市人民医院急救中心,吴忠 751100; 3.宁夏医科大学,银川 750004; 4.宁夏颅脑疾病重点实验室,银川 750004
Author(s):
REN Ping1ZHANG Helin2NIU Jianguo34LIU Yaqi34GU Jinhai34
1. Department of Neurosurgery, Wuzhong People's Hospital, Wuzhong 751100; 2.Emergency Center, Wuzhong People's Hospital, Wuzhong 751100; 3.Ningxia Medical University, Yinchuan, 750004; 4.Ningxia Key Laboratory of Cerebrocranial Diseases, Yinchuan 750004
关键词:
STAT3shRNA慢病毒
Keywords:
STAT3 shRNA lentivirus
分类号:
R394; Q78
DOI:
-
文献标志码:
A
摘要:
目的构建并鉴定靶向人STAT3基因的shRNA慢病毒载体。方法针对人STAT3基因设计有效靶序列,合成相应的shRNADNA寡核苷酸链;退火形成双链DNA后,与经过AgeI和EcoRI内切酶双酶切后的pGCSIL-GFP载体连接,产生shRNA慢病毒载体。PCR筛选阳性克隆,测序鉴定。以293T细胞包装生成完整病毒颗粒并测定其滴度。结果PCR鉴定与DNA测序证实合成的含STAT3shRNA慢病毒载体寡核苷酸链插入正确,浓缩病毒悬液的滴度为3×108TU·mL-1。结论成功构建了靶向人STAT3基因的shRNA慢病毒载体。
Abstract:
Objective To construct and identify a lentivirus-mediated short hairpin RNA(shRNA)vector targeting human signal transducer and activator of transcription 3(STAT3)gene. Methods According to the targeting sequence of human STAT3, the complementary shRNA DNA chains containing sense and anti-sense Oligo DNA fragments were designed, synthesized and cloned into the pGCSIL-GFP vector. The recombined lentiviral vector containing STAT3 shRNA was confirmed by PCR and DNA sequencing. The STAT3 shRNA vector was cotransfected with the packaging plasmids mixture into 293T cells by LipofectamineTM 2000. Virus in the supernatant was collected and the virus titer was measured. Results The PCR results and DNA sequencing demonstrated that the inserted sequences were correct. The titer of virus was 3×108TU·mL-1. Conclusion The lentivirus RNAi vector targeting STAT3 has been successfully constructed.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2013-07-16 基金项目:国家自然科学基金(81160312); 中国博士后科学基金面上项目(20100471492); 宁夏自然科学基金(NZ10193) 作者简介:任平,男,副主任医师,神经外科学。 通信作者:顾金海,男,副教授,神经生物学。E-mail:gjinhai@126.com
更新日期/Last Update: 2014-03-20