1. Enhanced expression of a recombinant malaria candidate vaccine in Escherichia coli by codon optimization.
- Author
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Zhou Z, Schnake P, Xiao L, and Lal AA
- Subjects
- Amino Acid Sequence, Animals, Antigens, Protozoan genetics, Base Sequence, Blotting, Western, Cell Division genetics, Chromatography, Agarose, Cloning, Molecular, DNA, Protozoan genetics, DNA, Protozoan immunology, Electrophoresis, Polyacrylamide Gel, Epitopes genetics, Epitopes immunology, Escherichia coli growth & development, Gene Expression drug effects, Gene Expression Regulation, Bacterial, Genes, Synthetic genetics, Genes, Synthetic immunology, Genetic Vectors genetics, Isopropyl Thiogalactoside pharmacology, Malaria Vaccines analysis, Malaria Vaccines genetics, Molecular Sequence Data, Plasmodium falciparum genetics, Protein Engineering methods, RNA, Transfer genetics, Sequence Alignment, Sequence Homology, Nucleic Acid, Vaccines, Synthetic analysis, Vaccines, Synthetic genetics, Codon genetics, Escherichia coli genetics, Gene Expression genetics, Malaria Vaccines biosynthesis, Plasmodium falciparum immunology, Vaccines, Synthetic biosynthesis
- 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.
- Published
- 2004
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