Abstract: Methods of increasing the cellular pool of A-CoA and thus driving the metabolic pathways in the direction of A-CoA containing metabolites by overexpressing rate limiting enzymes in A-CoA synthesis. Methods of increasing intracellular levels of CoA and A-CoA through genetic engineering of bacterial strains in conjunction with supplementation with precursor molecules.

Abstract: A process for assembling a series of DNA fragments generated by PCR into an ordered circular arrangement for replication and genetic work in cells. The PCR fragments are made with a modified nucleotide in the primers that can be removed with a DNA excision repair enzyme to generate a 3? overhang. The 3? overhangs are designed to allow directional annealing and thus sequential PCR fragments can be assembled by annealing the overhangs and subsequent ligation. Sequential addition of PCR fragments is facilitated by growing the chain on a solid support, and the assembled chain can be removed with a site specific recombinase if the first and last primers contain the recombinase site. The circularized assembled fragment can be directly used for cell transformation if the appropriate sequences are included, such as an origin of replication and a selectable marker.

Abstract: Methods of increasing the cellular pool of A-CoA and thus driving the metabolic pathways in the direction of A-CoA containing metabolites by overexpressing rate limiting enzymes in A-CoA synthesis. Methods of increasing intracellular levels of CoA and A-CoA through genetic engineering of bacterial strains in conjunction with supplementation with precursor molecules.

Abstract: A method of assembling large DNA fragments in a chromosome using site specific recombinases and alternating excisionases. The method may be performed in vitro or in vivo, but larger assemblies are possible when the assembly is performed in vivo. For an in vivo assembly, the cell must be engineered to contain the desired recombinases, each in an inducible construct so that the desired recombinase can be expressed at the correct time with the correct choice of inducing agent.

Abstract: Methods of increasing the cellular pool of A-CoA and thus driving the metabolic pathways in the direction of A-CoA containing metabolites by overexpressing rate limiting enzymes in A-CoA synthesis. Methods of increasing intracellular levels of CoA and A-CoA through genetic engineering of bacterial strains in conjunction with supplementation with precursor molecules.

Abstract: A method of assembling large DNA fragments in a chromosome using site specific recombinases and alternating excisionases. The method may be performed in vitro or in vivo, but larger assemblies are possible when the assembly is performed in vivo. For an in vivo assembly, the cell must be engineered to contain the desired recombinases, each in an inducible construct so that the desired recombinase can be expressed at the correct time with the correct choice of inducing agent.

Abstract: A method and related apparatus for cleansing or killing bacteria from raw oysters. The method involves exposing the meat, and possibly some natural fluids, of shucked oysters to high strength electric fields to cleanse or kill the bacteria present in and on the oyster. An apparatus to perform the method involves a sterilization tank made in part of two conductive plates spaced about one centimeter apart. The meat of shucked oysters is placed between the plates. A large voltage from energy-storing capacitors is applied to the conductive plates to create the electric field that kills the bacteria in the stomach and other cavities, and on the meat of the shucked oysters.