Abstract: An expression system for transforming E coll with a nucleic acid molecule of interest has an operator sequence of a cmt operon operatively linked to a promoter for the operator, and, a repressor sequence from a cym operon operatively linked to a promoter for the repressor. The expression system may have a nucleic acid molecule of interest, for example, a nucleic acid molecule that encodes a protein. Any type of E coll host cells may be transformed with the expression system. A method of producing a protein involves transforming an E coll host cell with the expression system having a nucleic acid molecule that codes for a protein, and, culturing the host cell in a culture medium under conditions in which the nucleic acid molecule will express the protein.

Abstract: Methylotrophic or methanotrophic bacteria such as Methylobacterium are transformed with a gene of interest, and expression of the gene is regulated by means of a cumate repressor protein and an operator sequence which is operatively linked to the gene of interest, and the addition of an external agent. Specifically, the cymR repressor and cmt operator from Pseudomonas putida may serve to regulate gene expression in methylotrophic or methanotrophic bacteria with the addition of cumate.

Abstract: Methylotrophic or methanotrophic bacteria such as Methylobacterium are transformed with a gene of interest, and expression of the gene is regulated by means of a cumate repressor protein and an operator sequence which is operatively linked to the gene of interest, and the addition of an external agent. Specifically, the cymR repressor and cmt operator from Pseudomonas putida may serve to regulate gene expression in methylotrophic or methanotrophic bacteria with the addition of cumate.

Abstract: An expression system for transforming E. coli with a nucleic acid molecule of interest has an operator sequence of a cmt operon operatively linked to a promoter for the operator, and, a repressor sequence from a cym operon operatively linked to a promoter for the repressor. The expression system may have a nucleic acid molecule of interest, for example, a nucleic acid molecule that encodes a protein. Any type of E. coli host cells may be transformed with the expression system. A method of producing a protein involves transforming an E. coli host cell with the expression system having a nucleic acid molecule that codes for a protein, and, culturing the host cell in a culture medium under conditions in which the nucleic acid molecule will express the protein.

Abstract: The integration of genes into methylobacterium, such as M. extorquens ATCC is disclosed, using a transposon system, preferably the mini-Tn7 transposon system, and under the control of a promoter, such as the strong methanol dehydrogenase promoter (PmxaF). Multicopy integration of genes of interest is also disclosed. The unique and specific attachment site for the Tn7 attachment (attTn7) is identified for M. extorquens.

Abstract: The integration of genes into methylobacterium, such as M. extorquens ATCC 55366 is disclosed, using a transposon system, preferably the mini-Tn7 transposon system, and under the control of a promoter, such as the strong methanol dehydrogenase promoter (PmxaF). Multicopy integration of genes of interest is also disclosed. The unique and specific attachment site for the Tn7 attachment (attTn7) is identified for M. extorquens.

Abstract: The present invention relates to a method of producing a recombinant peptide, a recombinant protein, or a product from metabolic engineering using a genetically modified methylotrophic bacterium, and more particularly to Methylobacterium extorquens ATCC 55366. The method comprises introducing an expression vector into the methylotrophic bacterium, the expression vector comprising a polynucleotide sequence, encoding for a peptide or protein, or allowing for production of a product from metabolic engineering under the control of a regulated promoter. The method also comprises growing the genetically modified methylotrophic bacterium in a minimal salts medium lacking organic sugars and containing methanol. A metal ion may be used for regulating the expression of the polynucleotide sequence by the promoter.

Abstract: The present invention relates to a method of producing a recombinant peptide, a recombinant protein, or a product from metabolic engineering using a genetically modified methylotrophic bacterium, and more particularly to Methylobacterium extorquens ATCC 55366. The method comprises introducing an expression vector into the methylotrophic bacterium, the expression vector comprising a polynucleotide sequence, encoding for a peptide or protein, or allowing for production of a product from metabolic engineering under the control of a regulated promoter. The method also comprises growing the genetically modified methylotrophic bacterium in a minimal salts medium lacking organic sugars and containing methanol. A metal ion may be used for regulating the expression of the polynucleotide sequence by the promoter.