Construction and expression of pBudCE4.1-SDF-1α-GDNF in bone marrow stromal cells of rhesus monkey
10.3760/cma.j.issn.1671-8925.2010.01.010
- VernacularTitle:外源性SDF-1α和GDNF基因在恒河猴骨髓基质细胞内的稳定表达
- Author:
Qiang XU
1
;
Bai-Nan XU
;
Ke LIU
;
Li PAN
;
Jun-Hua RAO
;
Ru-Xiang XU
Author Information
1. 解放军总医院
- Keywords:
Stromal cell-derived factor-1α;
Glial cell line-derived neurotrophic factor;
Bone marrow stromal cells;
Gene therapy
- From:
Chinese Journal of Neuromedicine
2010;9(1):43-46,50
- CountryChina
- Language:Chinese
-
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.
