Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (11): 1986-1989.doi: 10.3969/j.issn.1673-8225.2011.11.021

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Construction of recombinant DREAM-targeting small interfering RNA expressing plasmids

Chen Min1, Xiang Hong-bing2, Tian Yu-ke1   

  1. 1Department of Anesthesiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan  430030, Hubei Province, China
    2Department of Anesthesiology, Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou  510120, Guangdong Province, China
  • Received:2010-09-16 Revised:2010-10-12 Online:2011-03-12 Published:2011-03-12
  • Contact: Tian Yu-ke, Professor, Doctoral supervisor, Department of Anesthesiology, Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510120, Guangdong Province, China Yktian@tjh.tjmu.edu.cn
  • About author:Chen Min☆, Studying for doctorate, Attending Physician, Department of Anesthesiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China wuhan_chenmin@ sina.com

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Abstract:

BACKGROUND: DREAM is a multi-functional protein, which combine with different target proteins at different sites in cells. In vitro cultivate tests and animal experiments confirmed that DREAM is involving in onset mechanism of many different diseases.
OBJECTIVE: To construct recombinant DREAM-targeting small interfering RNA (siRNA) expressing plasmids. 
METHODS: Oligonucleotide containing the small hairpin of DREAM was designed and synthesized, which was inserted into the pDC316-EGFP-U6 plasmid double digested by BamHⅠ and Hind Ⅲ. The liation product was transformed competence E.coli DH5α. Positive clones were identified by PCR and sequencing. 
RESULTS AND CONCLUSION: The result of PCR and gene sequencing confirmed that the pDC316-EGFP-DREAMshRNA-U6 recombinant plasmid with 473 bp had been constructed successfully.

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