Simultaneous light emission from cloned Vibrio fischeri lux genes and expression of LamB or LamB-protein A fusion proteins in Escherichia coli depends on using a single chimeric operon.
Fusion proteins between LamB and immunoglobulin G binding domains of the Staphylococcus aureus protein A (SPA) have previously been shown to be located in the outer membrane and to convey immunoglobulin binding activity to intact Escherichia coli cells. However, the induced synthesis (tac promoter dependent) of these proteins severely impaired light production from the Vibrio fischeri lux operon present on a compatible plasmid and transcribed from its own control elements. Coordinate inducible expression of both phenomena, light emission and synthesis of LamB or LamB-SPA fusions, could be achieved by construction of artificial operons, joining all but luxl of the rightward lux operon to the 3' end of the LamB-spa expression cassettes, under transcriptional control of the tac promoter. Biotechnological applications are discussed. (c) 1995 John Wiley & Sons, Inc.
Steidler
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Laboratory of Molecular Biology, Universiteit Gent, K. L. Ledeganckstraat 35, B-9000 Gent, Belgium.
Fiers
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W
Remaut
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E