
A general method for solid-phase gene assembly on streptavidin-coated magnetic beads has been developed. The introduction of biotin in the 5'-end of the initiation oligonucleotide enables anchoring to the bead by means of the streptavidin-biotin interaction. The immobilization of one oligonucleotide enables controlled, stepwise annealing/ligation of successive 5'-phosphorylated oligonucleotides to rapidly build up predesigned gene constructs. In this report, we have assembled gene constructs of different lengths derived from the Plasmodium falciparum malaria blood-stage antigen Ag332. The encoded gene products were subsequently expressed in Escherichia coli using two parallel expression systems based on staphylococcal protein A and streptococcal protein G, respectively.
Base Sequence, Recombinant Fusion Proteins, Genes, Protozoan, Molecular Sequence Data, Plasmodium falciparum, Protozoan Proteins, Gene Expression, Antigens, Protozoan, DNA, Protozoan, Polymerase Chain Reaction, Animals, Amino Acid Sequence, Cloning, Molecular
Base Sequence, Recombinant Fusion Proteins, Genes, Protozoan, Molecular Sequence Data, Plasmodium falciparum, Protozoan Proteins, Gene Expression, Antigens, Protozoan, DNA, Protozoan, Polymerase Chain Reaction, Animals, Amino Acid Sequence, Cloning, Molecular
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