Symbol.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
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
Zheng Hong☆, Doctor, Attending physician, Department of Stomatology, Nanjing Children's Hospital of Nanjing Medical University, Nanjing 210009, Jiangsu Province, China [email protected]
Correspondence to: Ding Yin, Chief physician, Doctoral supervisor, Department of Orthodontics, Stomatology Hospital, Fourth Military Medical University of Chinese PLA, Xi'an 710032, Shaanxi Province, China [email protected]
Supported by: the National Natural Science Foundation of China, No. 30872913☆
Received: 2011-01-15
Accepted: 2011-03-29
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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, Hin dIII 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.
Zheng H, Li XH, Yin LH, Li J, Ding Y. Construction and identification of eukaryotic expression vector of RNA interference specific for estrogen receptor beta.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2011;15(19): 3459-3462. [http://www.crter.cnhttp://en.zglckf.com]
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