Construction of nervous specific promoter PDGF-EGFP vector and identification of tissue specificity
2009
Objective To construct an expression vector of enhanced green fluorescent protein(EGFP) containing nervous tissue-specific promoter-PDGF.Methods The PDGF-promoter sequence was amplified by PCR and the template was the genome of human blood cells.The PCR products were cloned into the pEGFP-1 vector and then pPDGF-EGFP vector was constructed.The vector was transfected into human neuroblastoma cells,SK-N-SH and four kinds of non-nervous recourse cells by lipofectamine.Expression of EGFP of transfected cells and tissue specificity of the vector were determined by transfection.Results The vector of pPDGF-EGFP was successfully constructed.After a transfection experiment,the EGFP was highly expressed in SKN-SH cells and scarcely expressed in the other four cells.Conclusion The vector of pPDGF-EGFP has a high nervous tissue specificity.It lays a foundation for studying target gene therapy of nervous system diseases.
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