Fusion reporter gene for bacterial luciferase

DETAILED DESCRIPTION The present invention comprises novel fusion genes useful as reporter genes, in particular, fusion genes which comprise two DNA sequences, a first DNA sequence which encodes an .
alpha.
-subunit of a bacterial luciferase and a second DNA sequence which encodes a .
beta.
-subunit of a bacterial luciferase.
The first and second DNA sequences are operably linked together to form a fusion gene which encodes a single protein with luciferase enzyme activity.
In particular, a luxA and luxB gene have been fused to form a novel luxAB gene.
The luxA and luxB gene sequences may be derived from a species of Vibrio bacterium.
Preferably, the sequences are derived from Vibrio harveyi.
In a particularly preferred embodiment, the luxA and luxB gene sequences are linked together by a third DNA sequence.
This third DNA sequence is preferably a restriction enzyme recognition site, most preferably an XbaI site (XbaI=TCTAGA).
The 5' to 3' order of the DNA sequences in a particularly preferred embodiment, that is the luxAB fusion gene described herein, is: 5'-luxA-TCTAGA-luxB-3'.
Those skilled in the art will recognize that the luxAB fusion gene is an important part of the present invention.
The luxA and luxB genes can be cloned from a Vibrio bacterium, in particular, Vibrio harveyi.
In addition, the sequence of the luxAB gene can be conventionally synthesized by the modified phosphotriester method using fully protected dioxyribonucleotide building blocks.
Such synthetic methods are well known in the art and can be carried out in substantial accordance with the procedure of Itakura et al.
, 1977, Science 198:1056 and Crea et al.
, 1978, Proc.
Natl.
Acad.
Sci.
USA 75:5765.
In addition, an especially preferred method is disclosed in Hsiung et al.
, 1983, Nucleic Acid Research 11:3227 and Narang et al.
, 1980, Methods in Enzymology 68:90.
In addition to the manual procedures referenced above, the luxAB fusion gene sequence can be synthesized using automated DNA synthesizers, such as the Systec 1450A or Applied Biosystems 380A DNA Synthesizers