Cloning and expressions of Renilla luciferase

• US 5,292,658 A
• Filed: 06/17/1993
• Issued: 03/08/1994
• Est. Priority Date: 12/29/1989
• Status: Expired due to Term

• Current Assignee: University of Georgia Research Foundation Incorporated The BOYD Graduate Studies Research Center
• Original Assignee: University of Georgia Research Foundation Incorporated The BOYD Graduate Studies Research Center
• Inventors: Lorenz, William W., Cormier, Milton J.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Moore, William W.
• Application Number:
  US08/079,700
• Time in Patent Office: 264 Days
• Field of Search: 435/69.1, 435/189, 435/252.3, 435/320.1, 536/23.2
• US Class Current: 435/252.33
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12Y 113/12005 Renilla-luciferin 2-monooxygenase (1.13.12.5), i.e. renilla-luciferase

Abstract

Genetic material encoding luciferase from the marine coelenterate Renilla has been isolated and characterized. This genetic material allows the production of peptides for use as labels in bioluminescence assays or can itself be directly used to identify luciferase genes from related organisms.

Claims

1. An isolated DNA or RNA molecule consisting essentially of a nucleotide sequence encoding a Renilla luciferase.

2. The molecule of claim 1, wherein said molecule consists essentially of luciferase coding sequence:

AGCTTAAAGATGACTTCGAAAGTTTATGATCCAGAACAAAGGAAACTTAT GATAACTGGTCCGCACTGGTGGGCCAGATGTAAACAAATGAATGTTCTTG ATTCATTTATTAATTATTATGATTCAGAAAAACATGCAGAAAATGCTGTT ATTTTTTTACATGGTAACGCGGCCTCTTCTTATTTATGGCGACATGTTGT GCCACATATTGAGCCAGTAGCGCGGTGTATTATACCAGATCTTATTGGTA TGGCAAATCAGGCAAATCTGGTAATGGTTCTTATAGGTTACTTGATCAT TACAAATATCTTACTGCATGGTTTGAACTTCTTAATTTACCAAAGAAGAT CATTTTTGTCGGCCATGATTGGGGTGCTTGTTTGGCATTTCATTATAGCT ATGAGCATCAAGATAAGATCAAAGCAATAGTTCACGCTGAAAGTGTAGTA GATGTGATTGAATCATGGGATGAATGGCCTGATATTGAAGAAGATATTGC GTTGATCAAATCTGAAGAAGGAGAAAAATGGTTTTGGAGAATAACTTCT TCGTGGAAACCATGTTGCCATCAAAAATCATGAGAAAGTTAGAACCAGAA GAATTTGCAGCATATCTTGAACCATTCAAAGAGAAAGGTGAAGTTCGTCG TCCAACATTATCATGGCCTCGTGAAATCCCGTTAGTAAAAGGTGGTAAAC CTGACGTTGTACAAATTGTTAGGAATTATAATGCTTATCTACGTGCAAGT GATGATTTACCAAAAATGTTTATTGAATCGGATCCAGGATTCTTTTCCAA TGCTATTGTTGAAGGCGCCAAGAAGTTTCCTAATACTGAATTTGTCAAAG TAAAAGGTCTTCATTTTTCGCAAGAAGATGCACCTGATGAAATGGGAAAA TATATCAAATCGTTCGTTGAGCGAGTTCTCAAAAATGAACAA or an essentially equivalent DNA or RNA sequence.

3. The molecule of claim 2, wherein said molecule is DNA.

4. The molecule of claim 3, wherein said molecule contains said luciferase sequence.

5. The molecule of claim 2, wherein said molecule is RNA and contains a sequence equivalent to said luciferase sequence.

6. An expression construct comprising,

a nucleotide sequence encoding a Renilla luciferase;

and a promoter sequence 5'"'"' to said nucleotide sequence, wherein said promoter sequence functions with said nucleotide sequence to express said nucleotide sequence in a host microorganism.

7. The molecule of claim 6, wherein at least one copy of said sequence is present in a recombinant DNA or RNA vector.

8. A genetically engineered microorganism, wherein said microorganism comprises the vector of claim 7, and is capable of producing biologically active luciferase.

9. The microorganism of claim 8, wherein said microorganism is an E. coli strain.