Recombinant human GDNF gene retroviral eukaryotic expression vector and expression.docVIP

 PAGE \* MERGEFORMAT 14 Recombinant human GDNF gene retroviral eukaryotic expression vector and expression Of: Chen Rui Xue-Ping Liu Zhijun Guo Chunhong [Abstract] Objective To construct containing glial cell line-derived neurotrophic factor (GDNF) and enhanced green fluorescent protein (EGFP retroviral vector, to import packaging cell PT67, detection of gene expression in cells. Methods PCR, GDNF gene amplification and IRES2 EGFP gene sequencing correctly connected with the retroviral vector pLXSN recombinant retroviral vector pLXSN IRES2 EGFP GDNF.EcoR I digested recombinant plasmid. liposome Lipofectamine 2000 mediated transfection packaging cell PT67, G418 screening and detection of virus titers. flow cytometry and fluorescence microscopy were used to detect transfection efficiency and gene expression. The results are about the size of PCR amplification, and 1 308 bp 558 bp specific band, sequencing confirmed the correct sequence of human GDNF sequence and EGFP. by EcoR I restriction enzyme digestion, recombinant plasmid pLXSN IRES2 EGFP GDNF. Fluorescence microscopy showed good expression of GDNF gene, the transfection efficiency by flow cytometry was 24.4%. conclusion is correct cloned human GDNF gene sequences and recombinant retroviral vector pLXSN IRES2 EGFP GDNF. PT67 packaging cells transfected by the good expression. [Keywords:] glial cell line-derived neurotrophic factor, retrovirus, liposome, gene transfer Parkinson’s disease (PD) poor clinical efficacy, there are scholars of bone marrow mesenchymal stem cells (MSCs transplanted into targeted mouse model of PD substantia nigra striatum and found that it can survive and differentiate into tyrosine hydroxylase (Tyrosine Hydroxylase, TH phenotype of neurons, thus improving the symptoms of PD in rats, but not prevent further nerve cell death. So some scholars to explore the neurotrophic factor in vitro transfection of target cells by means of stereotactic nigral transplantation into the mo