Improving heterologous gene expression in Aspergillus niger by introducing multiple copies of protein-binding sequence containing CCAAT to the promoter

Lett Appl Microbiol. 2003;36(6):358-61. doi: 10.1046/j.1472-765x.2003.01321.x.

Abstract

Aims: To increase the expression level of heterologous gene in Aspergillus niger.

Methods and results: Inserting multiple copies of an activator protein-binding site originally encoded in the cis-regulatory region of A. niger glucoamylase gene (glaA), CCAAT-containing sequence, into the promoter of an expression plasmid greatly enhanced the production of heterologous protein.

Conclusion: The promoter activity was increased by the introduction of the CCAAT-containing sequence resulting in increased gene expression.

Significance and impact of the study: This study provides evidence that the CCAAT-containing region of the glaA promoter of A. niger is a binding site for a positive transcriptional factor and demonstrates the possibility for improving the expression of both heterologous and endogenous genes in Aspergillus.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspergillus niger / genetics
  • Aspergillus niger / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Gene Dosage*
  • Gene Expression Regulation, Fungal*
  • Glucan 1,4-alpha-Glucosidase / chemistry*
  • Glucan 1,4-alpha-Glucosidase / genetics
  • Hemoglobins / genetics
  • Hemoglobins / metabolism*
  • Molecular Sequence Data
  • Plasmids / genetics
  • Promoter Regions, Genetic / genetics*
  • Recombination, Genetic
  • Transformation, Genetic
  • Truncated Hemoglobins

Substances

  • Bacterial Proteins
  • Hemoglobins
  • Truncated Hemoglobins
  • hemoglobin protein, Vitreoscilla
  • Glucan 1,4-alpha-Glucosidase