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Stem Cell Related Patent Number US6605449
| Title: | Synthetic ligation reassembly in directed evolution | | Inventors: | Short, Jay M.; Rancho Santa Fe, CA. USA | | Summary: | Described herein are methods of producing chimerized nucleic acid molecules composed of building-block sequences which are non-stochastically assembled to generate a display library. The invention relates to a process for the production of a library of mutagenized progeny polynucleotides from a progenitor polynucleotide template, wherein each of the 20 naturally encoded amino acids is encoded at each original codon position. Termed site-saturation mutagenesis, or simply saturation mutagenesis, this method is described as being preferably based on the use of the degenerate N,N,G/T sequence. Further disclosed are methods by which a non-stochastically produced library of chimeric nucleic acid molecules has an overall assembly order that is chosen by design, according to a set of progenitor templates, such as a family of esterase genes, or gene pathways such as encoding antibiotics, which can be shuffled to generate a sizable library of distinct progeny polynucleotide molecules and corresponding encoded polypeptides. Methods of screening the polynucleotide libraries are provided, enabling the identification of a desirable molecular species that has a desirable property, such as a specific enzymatic activity or a novel antibiotic. Applications in site-directed mutagenesis are included. | | Abstract: | | | US Patent Website: | Click Here for Full Text of Patent | | Title Number: | US6605449 | | Application Number: | US2000000594459 | | Date Filed: | 14/06/2000 | | Date Published: | 12/08/2003 | | Assignee: | Diversa Corporation, San Diego, CA, USA |
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