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| 小鼠Smad6基因RNAi慢病毒载体的构建与鉴定 | |
| 引用本文： | 刘铮,代继宏,符州,冯琳琳.小鼠Smad6基因RNAi慢病毒载体的构建与鉴定[J].生物学杂志,2011,28(5):19-23. |
| 作者姓名： | 刘铮 代继宏 符州 冯琳琳 |
| 作者单位： | 1. 儿童发育疾病研究省部共建教育部重点实验室 2. 重庆医科大学儿童医院呼吸中心 3. 儿科学重庆市重点实验室,重庆,400014 |
| 基金项目： | 重庆市科委自然科学基金资助 |
| 摘 要： | 构建小鼠Smad6基因RNA干扰（RNAi）慢病毒载体,有效沉默骨髓树突状细胞（BMDC）的Smad6基因表达,为构建骨髓致耐受DC用于哮喘等自身免疫疾病的研究。设计小鼠Smad6 shRNA序列,合成、退火,得到双链DNA,与经酶切后的Psih1-H1-copGFP shRNA Vector载体连接产生LV-shSmad6慢病毒载体,并测序鉴定。转染293TN细胞,包装产生慢病毒,测定滴度。感染小鼠骨髓树突细胞,检测Smad6基因的表达状况成功构建Smad6 shRNA的慢病毒载体LV-shSmad6。包装慢病毒,并显著抑制Smad6 mRNA水平及蛋白水平的表达。成功构建出小鼠Smad6基因shR-NA慢病毒载体,为后期研究Smad6基因在哮喘发病机制及新治疗方法提供了稳定的转染细胞载体。 |
| 关 键 词： | Smad6 RNA干扰 慢病毒载体 哮喘 |
Construction and identification of lentiviral vector of RNA interference of mouse Smad6 gene |
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| LIU Zheng,DAI Ji-hong,FU Zhou,FENG Lin-lin.Construction and identification of lentiviral vector of RNA interference of mouse Smad6 gene[J].Journal of Biology,2011,28(5):19-23. | |
| Authors: | LIU Zheng DAI Ji-hong FU Zhou FENG Lin-lin |
| Institution: | 1.Ministry of Education key Laboratory of Child Development and Disorders; 2.Center of Respiratory Diseases,Children′s Hospital of Chongqing Medical University; 3.Key Laboratory of Pediatrics in chongqing,CSTC 2009CA5002,Chongqing 400014,China) |
| Abstract: | To construct a lentiviral vector of RNA interference(RNAi) of mouse Smad6 gene and to effectively silence the Smad6 gene expression of bone marrow derived dendritic cells for construction the bone marrow tolerogenic dendritic cell for providing experimental foundation of athma of autoimmune disease,Smad6 shRNA sequence of mouse was designed by software.The complementary DNA containing both sence and antisence DNAs of the targeting sequence was designed,synthesized and cloned into the Psih1-H1-copGFP shRNA v... |
| Keywords: | Smad6 RNA interference lentiviral vector athma |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |