Composition for Analyzing Nucleic Acid

Abstract

A composition for analyzing nucleic acid that contains luciferase for which reactivity to dATP is equal to or less than 1/400 reactivity to ATP. A method for analyzing nucleic acid that comprises the use of the composition. A kit for analyzing nucleic acid comprising the composition. The method provides for an inexpensive, highly accurate and highly sensitive nucleic acid analysis that uses dATP instead of an expensive reagent having a low reactivity to DNA polymerase.

1 . A composition for analyzing nucleic acid, comprising a luciferase for which reactivity to dATP is equal to or less than 1/400 reactivity to ATP. 
     
     
         2 . The composition according to  claim 1 , wherein the amino acid at position 425 of luciferase is leucine; the amino acid at position 438 is glycine; the amino acid at position 532 is arginine; the amino acid at position 425 is leucine and the amino acid at position 438 is glycine; the amino acid at position 425 is leucine and the amino acid at position 532 is arginine; the amino acid at position 438 is glycine and the amino acid at position 532 is arginine; the amino acid at position 425 is leucine, the amino acid at position 438 is glycine and the amino acid at position 532 is arginine; the amino acid at position 344 is alanine, the amino acid at position 344 is valine, the amino acid at position 344 is isoleucine; or the amino acid at position 344 is alanine, the amino acid at position 425 is leucine and the amino acid at position 438 is glycine. 
     
     
         3 . The composition according to  claim 1 , wherein the luciferase is derived from  Luciola lateralis.    
     
     
         4 . The composition according to  claim 1 , wherein the luciferase is a chimeric luciferase that has an amino acid sequence of positions 1 to 448 of  Luciola cruciata  on the side of N terminal thereof, positions 217, 219 and 239 each being altered to isoleucine, and has an amino acid sequence of positions 447 to 450 of  Photinus pyralis  luciferase on the side of C terminal thereof. 
     
     
         5 . The composition for analyzing nucleic acid according to  claim 2 , wherein the lucerifase is derived from  Luciola lateralis.    
     
     
         6 . A method of analyzing nucleic acid comprising the use of the composition according to  claim 1 . 
     
     
         7 . A method of analyzing nucleic acid comprising the use of the composition according to  claim 2 . 
     
     
         8 . A method of analyzing nucleic acid comprising the use of the composition according to  claim 3 . 
     
     
         9 . A method of analyzing nucleic acid comprising the use of the composition according to  claim 4 . 
     
     
         10 . A method of analyzing nucleic acid comprising the use of the composition according to  claim 5 . 
     
     
         11 . A method of analyzing nucleic acid comprising the use of a composition comprising a mutant luciferase having the amino acid sequence of SEQ ID NO: 1, wherein said amino acid sequence has one or more mutations selected from the group consisting of 344V, 344I, 344A, and 344A+425L+438G, further wherein said mutant luciferase has reactivity to dATP which is equal to or less than 1/400 reactivity to ATP. 
     
     
         12 . A kit for analyzing nucleic acid comprising the composition according to  claim 1 . 
     
     
         13 . A kit for analyzing nucleic acid comprising the composition according to  claim 2 . 
     
     
         14 . A kit for analyzing nucleic acid comprising the composition according to  claim 3 . 
     
     
         15 . A kit for analyzing nucleic acid comprising the composition according to  claim 4 . 
     
     
         16 . A kit for analyzing nucleic acid comprising the composition according to  claim 5 . 
     
     
         17 . A kit for analyzing a composition comprising a mutant luciferase having the amino acid sequence of SEQ ID NO: 1, wherein said amino acid sequence has one or more mutations selected from the group consisting of 344V, 344I, 344A, and 344A+425L+438G, further wherein said mutant luciferase has reactivity to dATP which is equal to or less than 1/400 reactivity to ATP.