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- W2350797890 abstract "AIM To investigate the relationship between paraoxonase and the pathogenesis of cardiovascular diseases. METHODS BAC TO BAC baculovirus expression system to generate human serum paraoxonase(PON 1 isoform Q) was employed. PON 1 gene bearing EcoRⅠ site at upstream and HindⅢ site at downstream was amplified by PCR using the template of the full length PON 1 cDNA, sequenced and then cloned into the plasmid pFASTBACⅡ CD14 with 6× His·Tag to construct the baculovirus transfer vector pFASTBACⅡ PON 1. The recombinant transfer plasmid was transformed into the DH10BAC competent cells, and inserted into the Bacmid DNA by homologous recombination. The sf9 insect cells were infected with the recombinant infective baculovirus. Phenyl acetate was used as substrate to determine the PON 1 activity in the cultural supernatants. RESULTS PON 1 in the cultural supernatants was quantified at different multiplicity of infection(MOI) and time intervals. The best condition for expression was estimated at MOI=6-8, and the infection period of 96-108 h. The overall yield of PON 1 came to 4.5 mg per liter of supernatants. The result of Western blotting showed that the recombinant PON 1 can be recognized by anti 6×His monoclonal antibody. ESR also showed that the recombinant PON 1 could eliminate 55.9% of the lipid free radicals produced during lipid peroxidation of octadecadienoic acid. CONCLUSION The BAC TO BAC baculovirus mediated cells of Spodoptera frugiperda are suitable to express human PON 1 isoform Q in high efficiency." @default.
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- W2350797890 date "2003-01-01" @default.
- W2350797890 modified "2023-09-25" @default.
- W2350797890 title "Expression of human serum paraoxonase-1 isoform Q in baculovirus infected insect cells" @default.
- W2350797890 hasPublicationYear "2003" @default.
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