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Temperature-mediated regulation and downstream inducible selection for controlling gene expression from the bacteriophage lambda pL promoter.

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Affiliations

  • 1. Department of Chemistry, Purdue University, West Lafayette, IN 47907.
      Authors
    • Lowman HB 1
    (1 author)

Gene, 01 Nov 1990, 96(1):133-136
https://doi.org/10.1016/0378-1119(90)90353-s PMID: 2148296 

Abstract 

We have examined in detail the effects of various induction temperatures on the expression of a heterologous fusion gene controlled by the bacteriophage lambda PL promoter in a heat-inducible Escherichia coli expression system which utilizes the CIts857 repressor. Experiments performed over a temperature range spanning 29-42 degrees C indicate that, under our conditions, temperatures as low as 29 degrees C may be required to fully repress the CI857-controlled transcription from PL, and that the highest protein yields are obtained after induction at 36 degrees C for 6 h. We cloned the cat reporter gene downstream from a heterologous gene controlled by PL and found that cat expression at a low induction temperature permits the monitoring of productive transcription through the heterologous gene and thus aids in selecting transformants that are capable of producing the heterologous protein in E. coli.

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Read article at publisher's site: https://doi.org/10.1016/0378-1119(90)90353-s

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