Abstract: The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families.

Abstract: The present invention provides novel methods of generating a nucleic acid molecule. In certain embodiments, a double stranded nucleic acid chunk is generated from a ladder complex comprising partially complementary oligonucleotides, which chunk is combined with a nucleic acid acceptor molecule. In certain embodiments, the assembled chunk/nucleic acid acceptor molecule complex may be propagated in vivo or in vitro. The present invention also provides improved systems for generating a plurality of nucleic acid molecules that differ at one or more nucleotide positions. In certain embodiments, the plurality of nucleic acid molecules encodes a polypeptide or portion of a polypeptide.

Abstract: The present invention provides novel methods of generating a nucleic acid molecule. In certain embodiments, a double stranded nucleic acid chunk is generated from a ladder complex comprising partially complementary oligonucleotides, which chunk is combined with a nucleic acid acceptor molecule. In certain embodiments, the assembled chunk/nucleic acid acceptor molecule complex may be propagated in vivo or in vitro. The present invention also provides improved systems for generating a plurality of nucleic acid molecules that differ at one or more nucleotide positions. In certain embodiments, the plurality of nucleic acid molecules encodes a polypeptide or portion of a polypeptide.

Abstract: Systems are disclosed that are useful for introducing one or more targeted positions or regions of diversity into a nucleic acid molecule. In certain embodiments, diversity in a targeted position or region is generated by providing one or more degenerate primer sets and a template nucleic acid molecule, wherein the primers are extended in opposite directions against the template nucleic acid molecule in a polymerase-mediated extension reaction. In certain embodiments, the generated nucleic acid molecule into which diversity has been introduced comprises single-stranded regions at its termini, which single-stranded regions are capable of annealing to each other.

Abstract: The present invention provides a method for merging combinatorial biosynthesis with techniques of synthetic organic chemistry. In general, this method, combinatorial biology, involves 1) providing “starter units”, wherein the starter units are capable of being accepted by the modular biosynthetic enzymatic machinery, and have incorporated therein a “functional handle” capable of reacting with specific functionality present on a solid support; 2) feeding these “starter units” into the modular biosynthetic enzymatic machinery, in vivo or in vitro, to obtain complex template molecules; and 3) further functionalizing the complex template molecules using synthetic organic chemistry to provide a collection of complex “unnatural” natural products having structural, topological, stereochemical and functional diversity. In one preferred embodiment, the starter units are attached to solid support units prior to being exposed to the modular biosynthetic enzymatic machinery.

Abstract: The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families.

Abstract: The present invention provides novel methods of generating a nucleic acid molecule. In certain embodiments, a double stranded nucleic acid chunk is generated from a ladder complex comprising partially complementary oligonucleotides, which chunk is combined with a nucleic acid acceptor molecule. In certain embodiments, the assembled chunk/nucleic acid acceptor molecule complex may be propagated in vivo or in vitro. The present invention also provides improved systems for generating a plurality of nucleic acid molecules that differ at one or more nucleotide positions. In certain embodiments, the plurality of nucleic acid molecules encodes a polypeptide or portion of a polypeptide.

Abstract: The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families.

Abstract: The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families.

Abstract: The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families.