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[Construction of eukaryotic expression plasmid containing rat interleukin-10 gene and its expression in BRL cells in vitro].

[Article in Chinese]

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Department of Gastroenterology, Union Hospital, Fujian Medical University, Fuzhou 350001, China.



To construct eukaryotic expression vector of rat IL-10 gene and observe its expression in hepatocyte cell line BRL.


Total RNA was extracted from rat peripheral blood mononuclear cells. The full length coding region of IL-10 was amplified by RT- nested PCR and cloned into eukaryotic expression vector pcDNA3.0. The recombinant plasmid was transfected into BRL cells with either liposome Transfast or asialoglycoprotein receptor mediated liposome PEIjet-gal respectively. The expression of IL-10 mRNA was detected with PCR and that of IL-10 secreted from BRL cells transfected by liposome PEIjet-gal was detected with ELISA.


The recombinant plasmid was identified and confirmed with digestion of restriction endonuclease and DNA sequencing. Receptor mediated liposome PEIjet-gal exhibited significantly higher transfection efficiency than liposome Transfast and higher level secretory IL-10 expressed in BRL cells.


The eukaryotic expression vector of IL-10 gene was successfully constructed. Asialoglycoprotein receptor-mediated liposome had high transfection efficiency on hepatocytes, suggesting that it could be a potential hepatocyte-targeting delivery system for IL-10 gene therapy.

[Indexed for MEDLINE]

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