Solid-phase gene assembly of constructs derived from the Plasmodium falciparum malaria blood-stage antigen Ag332

Biotechniques. 1993 Mar;14(3):424-34.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigens, Protozoan / genetics*
  • Base Sequence
  • Cloning, Molecular / methods*
  • DNA, Protozoan / genetics
  • Gene Expression
  • Genes, Protozoan
  • Molecular Sequence Data
  • Plasmodium falciparum / genetics*
  • Plasmodium falciparum / immunology
  • Polymerase Chain Reaction
  • Protozoan Proteins / genetics
  • Recombinant Fusion Proteins / genetics

Substances

  • Antigens, Protozoan
  • DNA, Protozoan
  • Protozoan Proteins
  • Recombinant Fusion Proteins