Development of an α-amylase reporter system for efficient screening of clones with highly expressed heterologous protein in Hansenula polymorpha

Biotechnol Lett. 2010 Oct;32(10):1473-9. doi: 10.1007/s10529-010-0314-9. Epub 2010 May 22.

Abstract

To check feasibility and effectiveness of the α-amylase reporter system, two vectors were designed and tested using hepatitis B virus surface antigen (HBsAg) and Homo sapiens granulocyte-macrophage colony stimulating factor 2 (hGM-CSF2) as a model. By integrating the vector containing two independent cassettes into the same genome locus, high-producing clones of HBsAg (or hGM-CSF2) were screened using the α-amylase as a reporter. Results show there was a positive correlation (Correlation coefficient, R (2) > 0.95) between the yield of recombinant proteins and the α-amylase activity of corresponding transformants, which was independent of the gene dosage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotechnology / methods*
  • Genes, Reporter
  • Genetic Vectors
  • Granulocyte-Macrophage Colony-Stimulating Factor / biosynthesis
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics
  • Hepatitis B Surface Antigens / biosynthesis
  • Hepatitis B Surface Antigens / genetics
  • Humans
  • Mass Screening / methods
  • Pichia / enzymology*
  • Pichia / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • alpha-Amylases / biosynthesis*
  • alpha-Amylases / genetics

Substances

  • Hepatitis B Surface Antigens
  • Recombinant Proteins
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • alpha-Amylases