| Autor: |
Degelmann, Adelheid, Müller, Frank, Sieber, Heike, Jenzelewski, Volker, Suckow, Manfred, Strasser, Alexander W.M., Gellissen, Gerd |
| Předmět: |
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| Zdroj: |
FEMS Yeast Research; Aug2002, Vol. 2 Issue 3, p349, 13p |
| Abstrakt: |
The early status of strain development for the production of interleukin (IL)-6, IL-8, IL-10, and interferon (IFN) γ is described. The general approach to generating such strains was to amplify gene sequences encoding the mature forms of the various cytokines by PCR from commercially available cDNA sources. The design of the amplificates allowed an in-frame fusion to an MFα1 leader segment contained in two basic expression vectors, pFPMT121-MFα1 and pTPSMT-MFα1. The two vectors differ in that one harbors the methanol-inducible FMD promoter and the other the constitutive TPS1 promoter as control elements for heterologous gene expression. The most advanced process development example is that of IFNα-2a. Here, the MOX promoter derived from another key gene of methanol metabolism is used for expression control. The successful development of a production process for Hansenula polymorpha-derived IFNα-2a is summarized. This was achieved by combining genetic engineering of suitable production strains with improved processing capabilities for the secreted cytokine, and by purification procedures from cultures grown in yeast extract–peptone–glycerol-based media. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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