Formation of active bacterial luciferase between interspecific subunitsin vivo

✍ Scribed by Almashanu, S. ;Tuby, A. ;Hadar, R. ;Einy, R. ;Kuhn, J.

• Book ID: 102762114
• Publisher: John Wiley and Sons
• Year: 1995
• Weight: 898 KB
• Volume: 10
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170100304

✦ Synopsis

lnterspecific complementation between IuxAs and IuxBs from Vibrio harveyi, Vibrio fischeri. Photobacterium leiognathi and Xenorhabdus luminescens was examined in vivo. The individual genes from these species were cloned on different compatible plasmids or amplified by PCR and brought together t o yield cis combinations without extraneous DNA. The beta subunits from V. harveyi and X. luminescens form active enzyme only w i t h alpha subunits from one o f these species. All other combinations yield active enzymes. The lack o f activity o f the V. harveyi and X. luminescens beta subunits with the alpha subunits from V. fischeri and P. leiognathi results from a lack o f association. This was shown by in vivo competition in which these beta subunits were overproduced in comparison w i t h the beta and alpha o f V. fischeri. No reduction in light was found. Overall, t h e i n vivo results parallel those found in vitro using isolated denatured subunits and renaturation by removal o f the denaturant.