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

GLYCO-ENGINEERING: METHODS AND PROTOCOLS(2015)

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摘要
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.
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关键词
Disparity mutagenesis technique,N-linked oligosaccharide,Proofreading-deficient pol3,Therapeutic glycoproteins,Yeast
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