Abstract:

BACKGROUND: Recent studies have shown that pathogenesis of diabetic neurogenic bladder is associated with lack of nerve growth factor. Nerve growth factor (NGF) composed of three subunits is one of the most important neurotrophic factors. Beta subunit (beta-NGF) is the only biologically active subunit. Lentiviral vector is an ideal vector for gene therapy.
OBJECTIVE: To construct lentiviral expression vector carrying human beta-NGF gene.
METHODS: By the way of acquisition of the target gene, double restriction digestion of vector plasmid with Age Ⅰ/EcoR Ⅰ, connecting the target gene with vector plasmid, Beta-NGF gene was subcloned into the lentiviral vector plasmid pGC-FU, to generate the lentiviral expression vector plasmid, pGC-FU-beta-NGF. The cloning of human beta-NGF gene, and sequencing of the recombinant plasmid pGC-FU- beta-NGF were observed in the study.
RUSULTS AND CONCLUSION: For the recombinant lentiviral vector plasmid pGC-FU-β-NGF undergoing PCR, when actual PCR products share the same size with the expected, the recombinant plasmid is thought to have been successfully constructed. The result of sequencing showed the sequence of the cloned beta-NGF gene was consistent with that was reported in the GenBank. Plasmid pGC-FU- beta-NGF carried the correct beta-NGF gene.