Variant reverse transcriptase

摘要

The present invention provides a versatile mutant reverse transcriptase with high thermal stability, a nucleic acid thereof and a method for producing a mutant reverse transcriptase, a versatile kits for reverse transcription and detection, a method for improving thermal stability of a nucleic acid-related enzyme, which significantly improves thermal stability of a nucleic acid-related enzyme, and a reverse transcription method, which efficiently performs a reverse transcription. An amino acid residue in a nucleic acid interaction region of a wild-type enzyme is substituted with a positively-charged amino acid residue or a nonpolar amino acid residue, to form a nucleic acid interaction region having a positive effective charge larger than the nucleic acid interaction region of a wild-type enzyme.

主权项

1. A mutant reverse transcriptase comprising:
a DNA interaction region having a substitution of an amino acid residue corresponding to at least one of an aspartate residue at position 124 and a glutamate residue at position 286, such that either the aspartate residue at position 124, the glutamate residue at osition 286 or both the as sartate residue at position 124 and the glutamate residue at position 286 are substituted, in a DNA interaction region of a wild-type reverse transcriptase wherein the substitution or substitutions are that of a positively-charged amino acid residue or a nonpolar amino acid residue for the original negatively-charged amino acid residue or residues, and such that the mutant reverse transcriptase has a positive effective charge larger than that of the DNA interaction region in the wild-type reverse transcriptase, wherein the wild-type reverse transcriptase comprises an amino acid sequence corresponding to SEQ ID NO: 2, and wherein the mutant reverse transcriptase exhibits a reverse transcriptase activity.