Flavin reductase gene from Vibrio fischeri

• US 5,484,723 A
• Filed: 06/28/1994
• Issued: 01/16/1996
• Est. Priority Date: 06/28/1993
• Status: Expired due to Term

• Current Assignee: JNC Corporation (Chisso Corporation)
• Original Assignee: Chisso Corporation
• Inventors: Saigo, Kaoru, Zenno, Shuhei
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): HOBBS, LISA JOE
• Application Number:
  US08/266,570
• Time in Patent Office: 567 Days
• Field of Search: 435/189, 435/252.3, 435/69.1, 435/172.3, 435/320.1, 536/23.2
• US Class Current: 435/189
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12Y 114/14003 Alkanal monooxygenase FMN (1.14.14.3), i.e. bacterial-luciferase

Abstract

A flavin reductase gene is provided having the nucleotide sequence of SEQ ID NO:1, wherein the sequence is displayed in numbered triplets of capital letters, which numbers proceed sequentially from left to right and from the 5'"'"' terminus to the 3'"'"' terminus; the sequence of capital letters represent the purine and pyrimidine bases of the nucleotide sequence, as follows:

A is adenine; G is guanine; C is cytosine; T is thymine;

R is A or G; Y is T or C; N is A, T, C, or G; H is A, C, or T; and M is A or C;

wherein triplet number 237 thereof is TAA or TAG or TGA; and wherein further:

for triplets numbered 13, 21, 22, 36, 63, 85, 92, 111, 112, 126, 140, 150, 156, 186, 196, and 228, if the 3'"'"' nucleotide of a triplet is A or G, then the 5'"'"' nucleotide of the triplet is T or C, or if the 3'"'"' nucleotide of a triplet is T or C, then the 5'"'"' nucleotide of the triplet is C; and if the 5'"'"' nucleotide of a triplet is C, then the 3'"'"' nucleotide of the triplet is A, T, C, or G, or if the 5'"'"' nucleotide of a triplet is T, then the 3'"'"' nucleotide of the triplet is A or G;

for triplets numbered 19, 46, 56, 103,108, 123, 145, 206, and 214, if the 3'"'"' nucleotide of a triplet is T or C, then the 5'"'"' nucleotide of the triplet is A or C, or if the 3'"'"' nucleotide of a triplet is T or C, then the 5'"'"' nucleotide of the triplet is C; and if the 5'"'"' nucleotide of a triplet is G, then the 3'"'"' nucleotide of the triplet is A, T, C, or G, or if the 5'"'"' nucleotide of a triplet is A, then the 3'"'"' nucleotide of the triplet is A or G.

Claims

1. A flavin reductase gene having the nucleotide sequence of SEQ ID NO:

1, wherein the sequence is displayed in numbered triplets of capital letters, which numbers proceed sequentially from left to right and from the 5'"'"' terminus to the 3'"'"' terminus;

the sequence of capital letters represent the purine and pyrimidine bases of the nucleotide sequence, as follows;

A is adenine;

G is guanine;

C is cytosine;

T is thymine;

R is A or G;

Y is T or C;

N is A, T, C, or G;

H is A, C, or T; and

M is A or C;

wherein triplet number 237 thereof is TAA or TAG or TGA; and

wherein further;

for triplets numbered 13, 21, 22, 36, 63, 85, 92, 111, 112, 126, 140, 150, 156, 186, 196, and 228, if the 3'"'"' nucleotide of a triplet is A or G, then the 5'"'"' nucleotide of said triplet is T or C, or if the 3'"'"' nucleotide of a triplet is T or C, then the 5'"'"' nucleotide of said triplet is C; and

if the 5'"'"' nucleotide of a triplet is C, then the 3'"'"' nucleotide of said triplet is A, T, C, or G, or if the 5'"'"' nucleotide of a triplet is T, then the 3'"'"' nucleotide of said triplet is A or G;

for triplets numbered 19, 46, 56, 103, 108, 123, 145, 206, and 214, if the 3'"'"' nucleotide of a triplet is T or C, then the 5'"'"' nucleotide of said triplet is A or C, or if the 3'"'"' nucleotide of a triplet is T or C, then the 5'"'"' nucleotide of said triplet is C; and

if the 5'"'"' nucleotide of a triplet is G, then the 3'"'"' nucleotide of said triplet is A, T, C, or G, or if the 5'"'"' nucleotide of a triplet is A, then the 3'"'"' nucleotide of said triplet is A or G.

2. The flavin reductase gene according to claim 1, wherein said gene has the nucleotide sequence of SEQ ID NO:

3.

5. A recombinant vector comprising the nucleotide sequence of SEQ ID NO:

1, as set forth in claim 1.

6. The recombinant vector according to claim 5, wherein said SEQ ID NO:

1 is inserted into a plasmid vector.

7. A bacterium having a recombinant vector comprising the flavin reductase gene according to claim 1.

8. A process for producing an enzyme having the amino acid sequence of SEQ ID NO:

6, comprising cultivating a bacterium according to claim 7.

3. A flavin reductase gene having the nucleotide sequence of SEQ ID NO:

5.

4. A flavin reductase enzyme having the amino acid sequence of SEQ ID NO:

6.