Production of human neurotrophin-3 in baculovirus expression system

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 in...

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Veröffentlicht in:Chinese science bulletin 1998-09, Vol.43 (18), p.1562-1566
Hauptverfasser: Mu, Yuanyue, Liu, Hong, Zhang, Jie, Liu, Shuhong, Wu, Yan, Wang, Jiazheng, Fan, Ming
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Sprache:eng
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Zusammenfassung: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.
ISSN:1001-6538
2095-9273
1861-9541
2095-9281
DOI:10.1007/BF02883449