Disclosed is a method for promoting auto-induction of transcription of cloned
DNA in cultures of bacterial cells grown batchwise, the transcription being under the control of a
promoter whose activity can be induced by an exogenous
inducer whose ability to induce said
promoter is dependent on the
metabolic state of said bacterial cells. Initially, a culture media is provided which includes: i) an
inducer that causes induction of transcription from said
promoter in said bacterial cells; and ii) a
metabolite that prevents induction by said
inducer, the concentration of said
metabolite being adjusted so as to substantially preclude induction by said inducer in the early
stages of growth of the bacterial culture; but such that said
metabolite is depleted to a level that allows induction by said inducer at a later stage of growth. The culture medium is inoculated with a bacterial inoculum, the inoculum comprising bacterial cells containing cloned
DNA, the transcription of which is induced by said inducer. The culture is then incubated under conditions appropriate for growth of the bacterial cells. Also disclosed is a method for improving the production of a selenomethionine-containing
protein or polypeptide in a bacterial
cell, the
protein or polypeptide being produced by
recombinant DNA techniques, the bacterial
cell encoding a
vitamin B12-dependent
homocysteine methylase. The method for improving the production of this
protein or polypeptide includes culturing the bacterial
cell in a culture medium containing
vitamin B12. Finally, disclosed is a method for suppressing transcription of cloned
DNA in cultures of bacterial cells grown batchwise, said transcription being under the control of a promoter whose activity can be induced by an exogenous inducer whose ability to induce said promoter is dependent on the
metabolic state of said bacterial cells. This aspect includes the steps of: a) providing a culture medium comprising a
carbon source whose uptake and
metabolism by said bacterial cells suppresses induction of transcription from said promoter; b) inoculating the culture medium with a bacterial inoculum, the inoculum comprising bacterial cells containing cloned DNA, the transcription of which is suppressed by the
carbon source; and c) incubating the culture of step b), with shaking, under conditions appropriate for growth of the bacterial cells.