High density growth of T7 expression strains with auto-induction option
- Stony Brook, NY
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.
- Research Organization:
- Brookhaven National Laboratory (BNL), Upton, NY (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC02-98CH10886
- Assignee:
- Brookhaven Science Associates, LLC (Upton, NY)
- Patent Number(s):
- 7,560,264
- Application Number:
- 10/675,936
- OSTI ID:
- 1012613
- Country of Publication:
- United States
- Language:
- English
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High density growth of T7 expression strains with auto-induction option
High density growth of T7 expression strains with auto-induction option