FORMATION OF ACTIVE BACTERIAL LUCIFERASE BETWEEN INTERSPECIFIC SUBUNITS IN-VIVO

Interspecific complementation between luxAs and luxBs from Vibrio harv eyi. Vibrio fischeri, Photobacterium leiognathi and Xenorhabdos lumine scens was examined in vivo. The individual genes from these species we re cloned on different compatible plasmids or amplified by PCR and bro ught together to yield cis combinations without extraneous DNA. The be ta subunits from V. harveyi and X. luminescens form active enzyme only with alpha subunits from one of these species. All other combinations yield active enzymes. The lack of activity of the V. harveyi and X. l uminescens beta subunits with the alpha subunits from V. fischeri and P. leiognathi results from a lack of association. This was shown by in vivo competition in which these beta subunits were overproduced in co mparison with the beta and alpha of V. fischeri. No reduction in light was found. Overall. the in vivo results parallel those found in vitro using isolated denatured subunits and renaturation by removal of the denaturant.