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Stem Cell Related Patent Number US6537776
| Title: | Synthetic ligation reassembly in directed evolution | | Inventors: | Short, Jay M.; Encinitas, CA, USA | | Summary: | This invention introduces methods of non-stochastically producing a library of chimeric nucleic acid molecules. Described herein is a process for the rapid and facilitated production from a progenitor polynucleotide template of a library of mutagenized progeny polynucleotides wherein the assembly order is non-random such that 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 preferably based on the use of the degenerate N,N,G/T sequence. Methods are disclosed for the screening of the polynucleotide libraries such as to allow the identification of a desirable molecular species that has a desirable property, such as a specific enzymatic activity or a novel antibiotic. Further provided are methods of engineering genes and gene pathways by the introduction of regulatory sequences, such as promoters, that are operable in an intended host and which confer operability to a novel gene pathway when it is introduced into an intended host. Applications are included, such as in the screening of clones for specific activities of interest. | | Abstract: | | | US Patent Website: | Click Here for Full Text of Patent | | Title Number: | US6537776 | | Application Number: | US1999000332835 | | Date Filed: | 14/06/1999 | | Date Published: | 25/03/2003 | | Assignee: | Diversa Corporation, San Diego, CA, USA |
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