Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (19): 3459-3462.doi: 10.3969/j.issn.1673-8225.2011.19.009

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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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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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