Maxygen

About: Maxygen is a company organization based out in Redwood City, California, United States. It is known for research contribution in the topics: DNA shuffling & Nucleic acid. The organization has 260 authors who have published 234 publications receiving 18192 citations.

DNA shuffling of a family of genes from diverse species accelerates directed evolution

Abstract: DNA shuffling is a powerful process for directed evolution, which generates diversity by recombination, combining useful mutations from individual genes Libraries of chimaeric genes can be generated by random fragmentation of a pool of related genes, followed by reassembly of the fragments in a self-priming polymerase reaction Template switching causes crossovers in areas of sequence homology Our previous studies used single genes and random point mutations as the source of diversity An alternative source of diversity is naturally occurring homologous genes, which provide 'functional diversity' To evaluate whether natural diversity could accelerate the evolution process, we compared the efficiency of obtaining moxalactamase activity from four cephalosporinase genes evolved separately with that from a mixed pool of the four genes A single cycle of shuffling yielded eightfold improvements from the four separately evolved genes, versus a 270- to 540-fold improvement from the four genes shuffled together, a 50-fold increase per cycle of shuffling The best clone contained eight segments from three of the four genes as well as 33 amino-acid point mutations Molecular breeding by shuffling can efficiently mix sequences from different species, unlike traditional breeding techniques The power of family shuffling may arise from sparse sampling of a larger portion of sequence space

Methods for generating polynucleotides having desired characteristics by iterative selection and recombination

Abstract: A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

Methods and compositions for cellular and metabolic engineering

Abstract: The present invention is generally directed to the evolution of new metabolic pathways and the enhancement of bioprocessing through a process herein termed recursive sequence recombination. Recursive sequence recombination entails performing iterative cycles of recombination and screening or selection to “evolve” individual genes, whole plasmids or viruses, multigene clusters, or even whole genomes. Such techniques do not require the extensive analysis and computation required by conventional methods for metabolic engineering.

The Signal Recognition Particle

Abstract: The signal recognition particle (SRP) and its membrane-associated receptor (SR) catalyze targeting of nascent secretory and membrane proteins to the protein translocation apparatus of the cell. Components of the SRP pathway and salient features of the molecular mechanism of SRP-dependent protein targeting are conserved in all three kingdoms of life. Recent advances in the structure determination of a number of key components in the eukaryotic and prokaryotic SRP pathway provide new insight into the molecular basis of SRP function, and they set the stage for future work toward an integrated picture that takes into account the dynamic and contextual properties of this remarkable cellular machine.

Genome shuffling leads to rapid phenotypic improvement in bacteria

Abstract: For millennia, selective breeding, on the basis of biparental mating, has led to the successful improvement of plants and animals to meet societal needs1. At a molecular level, DNA shuffling mimics, yet accelerates, evolutionary processes, and allows the breeding and improvement of individual genes and subgenomic DNA fragments. We describe here whole-genome shuffling; a process that combines the advantage of multi-parental crossing allowed by DNA shuffling with the recombination of entire genomes normally associated with conventional breeding. We show that recursive genomic recombination within a population of bacteria can efficiently generate combinatorial libraries of new strains. When applied to a population of phenotypically selected bacteria, many of these new strains show marked improvements in the selected phenotype. We demonstrate the use of this approach through the rapid improvement of tylosin production from Streptomyces fradiae. This approach has the potential to facilitate cell and metabolic engineering and provide a non-recombinant alternative to the rapid production of improved organisms.