Evolution gene shuffling

Enzyme discovery and improvement high-throughput screening, biodiversity, directed evolution, gene shuffling, combinatorial chemistry, computational chemistry, bioinformatics, DNA microarray technology... 

Figure 3.2 Examples of gene shuffling methods used for DNA library creation in directed evolution, (a) Homology-dependent primer-independent DNA shuffling (b) homology-dependent primer-dependent StEP (c) homology-independent SHIPREC...

Other technologies include Molecular GRAMMR gene shuffling and molecular evolution technology and BAMF for pattern-recognition. 

Directed evolution involves multiple rounds of random mutation and selection combined with gene shuffling to evolve enzymes towards desired properties (reviewed in Arnold and Moore, 1997 Kuchner and Arnold, 1997). The group of Arnold has succeeded in evolving a dimethylformamide (DMF)-sensitive esterase for the cleavage of the loracarbef-/>-nitrobenzyl ester into an esterase that remains active in 15% DW (Moore et al, 1997). Most of the mutations that had been found in the solvent-resistant mutants could not have been predicted using current computational methods. 

The previous sections described structural studies of antibody maturation as a method to understand the evolution of binding (and catalysis) in the immune system. The technique of directed evolution parallels the process of affinity maturation. Both methods use random mutagenesis and gene shuffling, followed by screening and/or selection to identify mutants with the desired function. In contrast to affinity maturation,... 

Whereas site-directed mutagenesis is a technique that focuses on changing very specific residues based on the protein structure, directed evolution and gene shuffling take a more global approach. An advantage of these methods is the limited knowledge of the... 

It turns out that the huge variety of proteins is actually due largely to alternative splicing. The DNA does not make proteins directly each gene is first copied into RNA, a molecule similar to DNA but that codes in turn for the proteins directly. Since RNA sequences can be shuffled in various ways, this generates a variety of proteins, say four types, from each master gene. In billions of years of evolution the shuffled RNA molecules and resulting protein... 

---