Abstract:

To construct the recombinant adenovirus vector carrying the human gene neprilysin (NEP), obtain a high titer virus by packing and increasing, and explore the reducing of Aβ′s deposition to the damaged cells. MethodsA shuttle plasmid pAdeasyl was constructed in bacteria  and pAdNEP was generated. The replicationdeficient adenovirus was packed and propagated in the cell 293. Then the person′s nerve metrocyte lump cell SKNSH cells were infected  and NEP expression was  examined by RTPCR and Western blot. The endogenous damaged SKNSH cells were infected and the degeneration to Aβ was examined. ResultsAdNEP and control adenovirus AdGFP were successfully constructed. The recombinant adenovirus can efficiently infect the person′s nerve metrocyte lump cell SKNSH cells and can be used to examine high expression of NEP. The result of Elisa showed that high expression of NEP can reduce the extracellular deposition of Aβ. ConclusionHomologous recombination in bacteria is a simple and convenient strategy to generate adenovirus expressing exogenous genes. The nerve cells infected by ADNEP may reduce the deposition of Aβ through high expression of NEP.

Key words: Endopeptidases; Adenoviruses, human; Amyloid betaprotein; Alzheimer disease