Construction and expression of pBudCE4.1-SDF-1α-GDNF in bone marrow stromal cells of rhesus monkey / 中华神经医学杂志
Chinese Journal of Neuromedicine
; (12): 43-46,50, 2010.
Article
em Zh
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| ID: wpr-1032921
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ABSTRACT
Objective To observe the expression of stromal cell-derived factor-1α(SDF-1α)and glial cell line-derived neurotrophic factor(GDNF)genes in bone marrow stromal cells(BMSCs)of rhesus monkey. Methods With gene recombination technique,SDF-1α and GDNF genes obtained from cDNA were subcloned into pBudce4.1 vector to get pBudCE4.1-SDF-1-GDNF, evaluated by restriction enzyme analysis and sequencing analysis.The pBudCE4.1-SDF-1α-GDNF was transfected into BMSCs with lipofectamine2000.After 48 h,the expression of SDF-1α and GDNF was measured by RT-PCIL,Western blotting and immunohistochemistry. Results Correct construction of pBudCE4.1-SDF-1α-GDNF was identified by enzyme restriction analysis and sequencing analysis.Western blotting and immunohistochemistry confirmed the expressions Of SDF-1α and GDNF genes in the transfected cells.The protein expressions of GDNF and SDF-1α in the GDNF and SDF-1α transfected BMSCs were 6 times higher than those in the negative control cells. Conclusion The pBudCE4.1-SDF-1α-GDNF can successfully express SDF-1α and GDNF in BMSCs of rhesus monkey,which may be used in gene therapy for Parkinson's disease.
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Zh
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Chinese Journal of Neuromedicine
Ano de publicação:
2010
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Article