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抑制骨髓基质干细胞雌激素受体β基因表达的有效序列
引用本文:郑红,李秀华,殷丽华,李静,丁寅.抑制骨髓基质干细胞雌激素受体β基因表达的有效序列[J].中国组织工程研究与临床康复,2011,15(19):3459-3462.
作者姓名:郑红  李秀华  殷丽华  李静  丁寅
作者单位:1. 南京医科大学附属南京儿童医院口腔科,江苏省南京市,210009
2. 黑龙江省七台河市七煤集团公司总院口腔科,黑龙江省七台河市,154600
3. 黑龙江省富锦市中心医院口腔科,黑龙江省富锦市,156100
4. 解放军第四军医大学附属口腔医院正畸科,陕西省西安市,710032
基金项目:国家自然科学基金资助项目
摘    要:背景:雌激素受体β是否参与介导骨髓间充质干细胞的增殖与分化需进一步实验论证。目的:以RNAi技术寻找和验证对大鼠骨髓基质干细胞雌激素受体β基因表达抑制的有效序列。方法:根据GeneBank数据库提供的SD大鼠雌激素受体β基因核苷酸序列,选择设计能转录小发卡结构RNA(Small hairpin RNAs,shRNA)的DNA序列。再在两条互补碱基序列的5’端分别加上BamHⅠ(GATCC)和HindⅢ(AGCTT)酶的酶切位点,最后形成两条互补的克隆入pSilencer3.1-H1载体的发夹状siRNA模板序列,进行重组载体的碱基序列测定。结果与结论:重组质粒碱基序列鉴定后,证实真核表达载构建正确。雌激素受体β特异性siRNA真核表达载体构建成功。

关 键 词:雌激素受体β  RNA干扰  骨髓基质干细胞  siRNA真核表达载体  基因

Construction and identification of eukaryotic expression vector of RNA interference specific for estrogen receptor beta
Zheng Hong,Li Xiu-hua,Yin Li-hua,Li Jing,Ding Yin.Construction and identification of eukaryotic expression vector of RNA interference specific for estrogen receptor beta[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(19):3459-3462.
Authors:Zheng Hong  Li Xiu-hua  Yin Li-hua  Li Jing  Ding Yin
Institution:1Department of Stomatology,Nanjing Children’s Hospital of Nanjing Medical University,Nanjing 210009,Jiangsu Province,China;2Department of Stomatology,Qimei Group Company,Qitaihe 154600,Heilongjiang Province,China;3Department of Stomatology,Central Hospital of Fujin City,Fujin 156100,Heilongjiang Province,China;4Department of Orthodontics,Stomatology Hospital,Fourth Military Medical University of Chinese PLA,Xi’an 710032,Shaanxi Province,China
Abstract:BACKGROUND: Whether estrogen receptor beta (ER-β) participates to mediate proliferation and differentiation of bone marrow mesenchymal stem cells is unclear.OBJECTIVE: To construct and identify a specific small interfering RNA (siRNA) suppressing estrogen receptor beta gene.METHODS: Genome sequences of ER-β gene was retrieved from Genbank and cDNA was designed coding expression of small hairpin RNAs (shRNA) for ER-β gene.It was cloned into the expression vector pSilencer 3.1-H1 into a short hairpin RNA with BamH Ⅰ,HindⅢ sticky end,terminated cognition sequence and a loop structure,followed by being cloned in pRNAT-U6.1 /Neo to construct a expression vector of ER-β-specific siRNA.At last the vector was evaluated by enzyme cutting and gene sequencing test.RESULTS AND CONCLUSION: The recombinant plasmid of RNA interference specific for ER-β identified by the restriction map completely coincided with the designs.The siRNA eukaryotic expression vector against ER-β mRNA has been successfully constructed.
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