Luciferases
First Claim
1. An isolated protein having luciferase activity which protein has over 60% amino acid sequence homology to the luciferase from Photinus pyralis, Luciola mingrelica, Luciola cruciata, or Luciola lateralis and includes the amino acid sequence XGDDKPGA wherein X is an amino acid residue other than glutamate, glycine, proline or aspartic acid.
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Accused Products
Abstract
Proteins are provided having luciferase activity with greater heat stability than wildtype luciferases by replacing the glutamate equivalent to that at position 354 of Photinus pyralis luciferase or 356 of Luciola luciferases with an alternative amino acid, particularly lysine. DNA, vectors and cells that encode for and express the proteins are also provided as are test kits and reagents for carrying out luminescence assays using the proteins of the invention. Preferred proteins have a second replaced amino acid at a position equivalent to position 215 of Photinus pyralis luciferase or 217 of Luciola luciferases.
37 Citations
32 Claims
- 1. An isolated protein having luciferase activity which protein has over 60% amino acid sequence homology to the luciferase from Photinus pyralis, Luciola mingrelica, Luciola cruciata, or Luciola lateralis and includes the amino acid sequence XGDDKPGA wherein X is an amino acid residue other than glutamate, glycine, proline or aspartic acid.
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26. A method of increasing the heat stability of a protein having luciferase activity, which protein has over 60% amino acid sequence homology to the luciferase from Photinus pyralis, Luciola mingrelica, Luciola cruciata, or Luciola lateralis, and comprises an amino acid sequence XGDDKPGA, said method comprising replacing X with an amino acid, analogue or modification thereof which is different from glutamate, glycine, proline, or aspartic acid.
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27. In a luciferase which has over 60% amino acid sequence homology to the luciferase from Photinus pyralis, Luciola mingrelica, Luciola cruciata, or Luciola lateralis, is thermally stable to a temperature of about 30°
- C. and comprises an amino acid sequence XGDDKPGA, the improvement comprising an amino acid other than glutamate, glycine, proline or aspartic acid as a replacement for X.
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28. An isolated and purified protein comprising the amino acid sequence of SEQ ID NO:
- 2 wherein Xaa is one of tryptophan, valine, leucine, isoleucine, asparagine, lysine and arginine or an analogue or modification of these.
- View Dependent Claims (29, 30, 31, 32)
Specification