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PubMed · 12903444

[Construction and expression of eukaryotic expression vector pBK-IL-1ra].

G Yang·F Li·M Jiang·C Di·J Huang
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Abstract

OBJECTIVE: Construction of eukaryotic vector with high expression of interleukin-1 receptor antagonist (IL-1ra) for IL-1ra gene delivery. METHODS: (1) Eukaryotic expression vector pBK-IL-1ra was constructed by recombinant DNA technique, and identified by restriction mapping enzymes, PCR, and DNA sequence analysis. (2) Expressions of the IL-1ra mRNA and IL-1ra in vitro and in vivo were detected by in situ hybridization, ELISA, and immunohistochemistry. RESULTS: (1) The correct construction of pBK-IL-1ra was identified by the method above. (2) The higher levels of IL-1ra in the supernate of COS-7 cells (24 h, (P < 0.001; 48 h, P < 0.01) and the over expression of IL-1ra mRNA (P < 0.01) and IL-1ra (P < 0.05) in the synoviocytes have been detected after 48 h of the transfection with pBK-IL-1ra. (3) The over expression of IL-1ra in muscle of the mice was detected by immunohistochemistry after 4 days of pBK-IL-1ra injection (P < 0.01) CONCLUSIONS: We have successfully constructed eukaryotic expression vector pBK-IL-1ra which could highly expressed IL-1ra in vitro and in vivo, and it offers a novel possibility in the research of IL-1ra gene therapy.

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AnimalsCOS CellsGene ExpressionGene Transfer TechniquesGenetic VectorsHumansInterleukin 1 Receptor Antagonist ProteinMiceMice, Inbred DBAPlasmidsSialoglycoproteinsU937 CellsEnglish AbstractJournal ArticleResearch Support, Non-U.S. Gov't

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G Yang, F Li, M Jiang, C Di, J Huang. 2000. [Construction and expression of eukaryotic expression vector pBK-IL-1ra].. https://pubmed.ncbi.nlm.nih.gov/12903444/

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