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Development of a Valuable Yeast Strain Using a Novel Mutagenesis Technique for the Effective Production of Therapeutic Glycoproteins

  • Hiroko Abe
  • Ken-ichi Nakayama
Part of the Methods in Molecular Biology book series (MIMB, volume 1321)

Abstract

The so-called disparity mutagenesis technique selectively elevates mutation in the lagging strand of DNA by using a mutant form of DNA polymerase δ, encoded on a proofreading-deficient pol3 gene. This novel mutagenesis technique generates a pool of mutants that includes a no-mutant strain together with mutant strains carrying multiple mutations. By using a suitable screening system it is possible to isolate the desired mutant strain from this pool of mutants. Here, we used our novel mutagenesis technique to isolate a yeast strain with good growth characteristics that was glycosylation deficient.

Key words

Disparity mutagenesis technique N-linked oligosaccharide Proofreading-deficient pol3 Therapeutic glycoproteins Yeast 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Hiroko Abe
    • 1
  • Ken-ichi Nakayama
    • 2
  1. 1.Health Research InstituteNational Institute of Advanced Industrial Science and Technology (AIST)TakamatsuJapan
  2. 2.Research Institute for Sustainable ChemistryNational Institute of Advanced Industrial Science and Technology (AIST)Higashi-HiroshimaJapan

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