Abstract: Enhanced resistance to glyphosate, an inhibitor of the aromatic amino acid biosynthesis pathway, is imparted to a glyphosate sensitive host. A mutated aroA gene is employed which expresses 5-enolpyruvyl-3- phosphoshikimate synthase (EC: 2.5.1.19) (ES-3-P synthase). Methods are provided for obtaining the aroA mutation which provides the enzyme resistant to inhibition by glyphosate, means for introducing the structural gene into a sensitive host, as well as providing a method of producing the enzyme.The E. coli strain C600(pPMG1) has been deposited at the A.T.C.C. on Dec. 14, 1982 and given A.T.C.C. Accession No. 39256.
Abstract: Method and compositions are provided for introducing alien DNA into a nucleic acid in vivo by providing a plasmid capable of replication in a first host, which is capable of conjugation with a second host into which the alien DNA is to be introducted. The plasmid is characterized by being capable of maintenance in said first host, being incapable of maintenance in said second host, having a region of homology with DNA present in said second host, not self-transmissable but capable of mobilization by a helper plasmid and having a marker, which may be the alien DNA, for selection in said second host. The invention finds particular application for introducing alien DNA into the Ti plasmid of Agrobacterium tumefaciens for introduction into plant cells.
Abstract: Microinjection techniques for plant protoplasts utilize a holding pipette for immobilizing the protoplast while an injection pipette is utilized to inject the macromolecule. In order to manipulate the protoplasts without damage, the protoplasts are cultured for from about 1 to 5 days before the injection is performed to allow for partial regeneration of the cell wall. It was found that injection through the partially regenerated cell wall could still be accomplished and particular compartments of the cell could be targeted. The methods are particularly useful for transformation of plant protoplasts with exogenous genes.
Abstract: A method and assay kit for determining the presence of an enzyme in a sample are provided. A sample suspected of containing the enzyme is applied to an image gel, which may be any conventional material, typically agarose gel. The sample may first be subjected to electrophoresis, or may be detected directly using the present invention. The image gel includes an immobilized phase capable of binding a product of the enzyme but not the substrate. By exposing the enzyme to substrate, and drawing the resulting reaction mixtures through the image gel, only the product is bound in the image gel. The presence of enzyme may then be determined by detecting the product in the image gel. In addition to developing electrophoresis gels, the method of the present invention will find great use in screening a plurality of complex mixtures which have been separated by other conventional techniques.
Abstract: Beer production is improved by the introduction of oxalate decarboxylase during mashing and/or fermentation. The addition of the oxalate decarboxylase diminishes or eliminates calcium oxalate precipitation as a deposit on tank walls and equipment.
Abstract: Enhanced resistance to glyphosate, an inhibitor of the aromatic amino acid biosynthesis pathway, is imparted to a glyphosate sensitive host. A mutated aroA gene is employed which expresses 5-enolpyruvyl-3-phosphoshikimate synthetase (EC: 2.5.1.19) (ES-3-P synthetase). Methods are provided for obtaining the aroA mutation which provides the enzyme resistant to inhibition by glyphosate, means for introducing the structural gene into a sensitive host, as well as providing a method of producing the enzyme.The E. coli strain C600(pPMG1) has been deposited at the A.T.C.C. on Dec. 14, 1982 and been given A.T.C.C. accession no. 39256.