Abstract
Human NT-3 gene was inserted into pPAK9 and the resultant plasmid was used to co-transfect sf21 cells with Bsu36 I digested Bac6 genomic DNA. Recombinant baculovirus was isolated by plaque assay. Slot-hybridization, immunocytochemistry and serum-free culture of DRG demonstrated that NT-3 had been integrated into Bac6 DNA, expressed and secreted. The yield of purified hNT-3 was 200 μg from 200 mL cultural supernatant after passing CM Sepharose Fast Flow column, the concentration for its maximum bioactivity was 4 ng/mL.
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