Sulfurylase-luciferase fusion proteins and thermostable sulfurylase

• US 6,902,921 B2
• Filed: 05/23/2002
• Issued: 06/07/2005
• Est. Priority Date: 10/30/2001
• Status: Expired due to Fees

• Current Assignee: 454 Life Sciences Corporation (Roche Holding AG)
• Original Assignee: 454 CORPORATION
• Inventors: Srinivasan, Maithreyan, Reifler, Michael
• Primary Examiner(s): Slobodyansky, Elizabeth
• Application Number:
  US10/154,515
• Publication Number:
  US 20030113747A1
• Time in Patent Office: 1,111 Days
• Field of Search: 435/194, 435/69.7, 435/189, 435/193, 536/23.2, 536/23.4
• US Class Current: 435/194
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12N 9/1241 Nucleotidyltransferases (2.7.7)

Abstract

The present invention relates to the field of DNA recombinant technology. More specifically, this invention relates to fusion proteins comprising an ATP generating polypeptide joined to a polypeptide that converts ATP into a detectable entity. Accordingly, this invention focuses on sulfurylase-luciferase fusion proteins. This invention also relates to pharmaceutical compositions containing the fusion proteins and methods for using them.

Claims

1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

2, 4 and 6.

2. The polypeptide of claim 1 wherein the polypeptide comprises SEQ ID NO:

2.

3. The polypeptide of claim 1 attached to a mobile support.

4. The polypeptide of claim 3 wherein the polypeptide is attached to the mobile support by a covalent or non-covalent interaction.

5. The polypeptide of claim 4 wherein the polypeptide is attached by a linkage selected from the group consisting of a CO²+-hexahistidine linkage, a Ni²⁺-hexahistidine linkage a biotin binding protein/biotin linkage, a glutathione S-transferase/glutathione linkage, a monoclonal antibody/antigen linkage, a maltose binding protein/maltose linkage and pluronic linkage.

6. The polypeptide of claim 5 wherein the biotin binding protein is selected from the group consisting of streptavidin and avidin.

7. The polypeptide of claim 3 wherein the mobile support is selected from the group consisting of a bead, optical fiber and glass surface.

8. The polypeptide of claim 7 wherein the bead is a nickel-agarose bead or a magnetic porous glass Streptavidin bead.