PCR-Based Gene Synthesis to Produce Recombinant Proteins for Crystallization

PCR-Based Gene Synthesis to Produce Recombinant Proteins for Crystallization

Docket: UAH-P-08002


UAH researchers have developed a Polymerase Chain Reaction (PCR)-based gene synthesis technique coupled to homologous recombination in vivo to quickly construct genes without the use of any additional enzymes. This invention allows the researchers to carry out the easy and feasible gene synthesis projects in their home laboratories without the need of intricate and complex manipulations or purifications. This is because it avoids the need of additional enzymes. It also facilitates the investigation of the protein structures, since it combines the feasible gene synthesis techniques with quick cloning methods.

Using this method, the entire procedure from gene to crystal screening can be performed within two weeks. This technique integrates gene synthesis, the DNA error correction, and subcloning techniques into a non-automated gene to crystal pipeline. Genes can be designed, synthesized, and implemented for recombinant expression of protein crystallization. In other words, it allows the use of the current methods of gene assembly for recombinant protein (artificially-built protein) production.

The PCR-based gene synthesis procedure is based on sequential assembly such that homogenous DNA products can be obtained after each synthesis step. Experiments were performed on 18 different genes and gene fragments, confirming the reproducibility of this method.


  • Protein engineering
  • Gene synthesis


  • More robust than conventional Thermodynamically Balanced Inside-Out (TBIO) methods, because the steps performed are simpler and require less manipulation
  • No automation required
  • No extensive purification required
  • No increase in cost
  • Faster than conventional methods (the whole procedure from gene to crystal screening can be done in two weeks)
  • The result is a reliable and consistent DNA assembly for almost any sequence


  • State of Development: Prototype
  • Licensing Status: Available for licensing
  • Patent Status: Proprietary