中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (19): 3459-3462.doi: 10.3969/j.issn.1673-8225.2011.19.009

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

抑制骨髓基质干细胞雌激素受体β基因表达的有效序列

郑  红1，李秀华2，殷丽华3，李  静1，丁  寅4   

  1. 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
  • 收稿日期:2011-01-15 修回日期:2011-03-29 出版日期:2011-05-07 发布日期:2011-05-07
  • 通讯作者: 丁寅，主任医师，博士生导师，解放军第四军医大学口腔医学院正畸科，陕西省西安市710032 dingyin@fmmu.edu.cn
  • 作者简介:郑红☆，女，1975年出生，黑龙江省绥滨县人，汉族，2008年解放军第四军医大学毕业，博士，主治医师，主要从事口腔正畸骨组织改建的研究。 donkeyzh@163.com
  • 基金资助:

    国家自然科学基金资助项目(30872913)。

Construction and identification of eukaryotic expression vector of RNA interference specific for estrogen receptor beta

Zheng Hong1, Li Xiu-hua2, Yin Li-hua3, Li Jing1, Ding Yin4   

  1. 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
  • Received:2011-01-15 Revised:2011-03-29 Online:2011-05-07 Published:2011-05-07
  • Contact: Ding Yin, Chief physician, Doctoral supervisor, Department of Orthodontics, Stomatology Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China dingyin@fmmu.edu.cn
  • About author:Zheng Hong☆, Doctor, Attending physician, Department of Stomatology, Nanjing Children’s Hospital of Nanjing Medical University, Nanjing 210009, Jiangsu Province, China donkeyzh@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30872913*

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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BamH Ⅰ(GATCC)和Hind Ⅲ (AGCTT)酶的酶切位点，最后形成两条互补的克隆入pSilencer 3.1-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&beta, RNA干扰, 骨髓基质干细胞, siRNA真核表达载体, 基因

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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