Luciferase gene and novel recombinant DNA as well as a method of producing luciferase

• US 4,968,613 A
• Filed: 07/26/1988
• Issued: 11/06/1990
• Est. Priority Date: 07/29/1987
• Status: Expired due to Term

• Current Assignee: Kikkoman Corporation
• Original Assignee: Kikkoman Corporation
• Inventors: Masuda, Tsutomu, Nakano, Eiichi, Tatsumi, Hiroki
• Primary Examiner(s): NOT, DEFINED
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US07/224,445
• Time in Patent Office: 833 Days
• Field of Search: 435/68, 435/70, 435/235, 435/172.3, 435/320, 435/849, 435/253, 435/320, 435/272, 435/189, 435/69.1, 536/27, 935/14, 935/60, 935/29
• US Class Current: 435/189
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12Y 113/12007 Photinus-luciferin 4-monooxygenase (ATP-hydrolysing) (1.13.12.7), i.e. firefly-luciferase

Abstract

A luciferase gene isolated from Luciola cruciata (Japanese firefly) coding for an amino acid sequence shown in FIG. 4 and a novel recombinant DNA characterized by incorporating a gene coding for luciferase into a vector DNA are disclosed. There is also disclosed a method of producing luciferase which comprises culturing in a medium a microorganism containing a recombinant DNA having inserted a gene coding for luciferase in a vector DNA and belonging to the genus Escherichia capable of producing luciferase and collecting luciferase from the culture.

Claims

1. An isolated luciferase gene coding for an amino acid sequence shown in FIG. 4.

2. An isolated luciferase gene according to claim 1, having a DNA sequence of FIG. 3.

3. A recombinant DNA comprising an isolated luciferase DNA from Luciola cruciata.

4. A recombinant DNA according to claim 3 wherein said isolated luciferase DNA has a nucleotide sequence shown in FIG. 3.

5. A recombinant DNA according to claim 3 wherein said isolated luciferase DNA codes for an amino acid sequence of FIG. 4.

6. A recombinant DNA according to claim 3, wherein said isolated luciferase DNA from Luciola cruciata is inserted into plasmid pUC18 DNA.

7. A method of producing luciferase which comprises culturing, in a medium, a microorganism belonging to the genus Escherichia capable of producing luciferase, said microorganism containing a recombinant DNA having inserted therein an isolated luciferase gene from Luciola cruciata and collecting luciferase from the culture.

8. A method of producing luciferase according to claim 7, wherein said gene codes for an amino acid sequence shown in FIG. 4.

9. A method of producing luciferase according to claim 7 wherein said gene has a nucleotide sequence shown in FIG. 3.

10. A method for producing luciferase according to claim 7, wherein said recombinant DNA is one produced by inserting an isolated luciferase gene from Luciola cruciata into pUC18 DNA.