Enhanced expression of a recombinant malaria candidate vaccine in Escherichia coli by codon optimization

Protein Expr Purif. 2004 Mar;34(1):87-94. doi: 10.1016/j.pep.2003.11.006.

Abstract

This study was conducted to compare the expression of three constructs of a multistage candidate vaccine (FALVAC-1) against Plasmodium falciparum in an Escherichia coli system: a synthetic gene with P. falciparum codons, a synthetic gene with optimized E. coli codons, and a synthetic gene with P. falciparum codons co-transformed with a RIG plasmid, which encodes three tRNAs (AG(A/G), ATA, GGA) that recognize rare E. coli codons. The expression of the protein increased at least threefold with codon optimization. The presence of the RIG plasmid in the co-transforming cells did not significantly increase the expression level of the gene with P. falciparum codons. The growth of cells transformed by the construct with P. falciparum codons was significantly slower than that of cells transformed by the construct with optimized E. coli codons after induction of protein expression with IPTG. The cells containing the non-codon optimized gene co-expressed with RIG plasmid had the slowest growth at all time points in culture. Thus, codon optimization significantly increases the yield of P. falciparum candidate vaccines in the E. coli expression system.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigens, Protozoan / genetics
  • Base Sequence
  • Blotting, Western
  • Cell Division / genetics
  • Chromatography, Agarose
  • Cloning, Molecular
  • Codon / genetics*
  • DNA, Protozoan / genetics
  • DNA, Protozoan / immunology
  • Electrophoresis, Polyacrylamide Gel
  • Epitopes / genetics
  • Epitopes / immunology
  • Escherichia coli / genetics*
  • Escherichia coli / growth & development
  • Gene Expression / drug effects
  • Gene Expression / genetics*
  • Gene Expression Regulation, Bacterial
  • Genes, Synthetic / genetics
  • Genes, Synthetic / immunology
  • Genetic Vectors / genetics
  • Isopropyl Thiogalactoside / pharmacology
  • Malaria Vaccines / analysis
  • Malaria Vaccines / biosynthesis*
  • Malaria Vaccines / genetics
  • Molecular Sequence Data
  • Plasmodium falciparum / genetics
  • Plasmodium falciparum / immunology*
  • Protein Engineering / methods
  • RNA, Transfer / genetics
  • Sequence Alignment
  • Sequence Homology, Nucleic Acid
  • Vaccines, Synthetic / analysis
  • Vaccines, Synthetic / biosynthesis*
  • Vaccines, Synthetic / genetics

Substances

  • Antigens, Protozoan
  • Codon
  • DNA, Protozoan
  • Epitopes
  • Malaria Vaccines
  • Vaccines, Synthetic
  • Isopropyl Thiogalactoside
  • RNA, Transfer