High fidelity reverse transcriptases and uses thereof
AssigneeInvitrogen Corporation
InventorsRobert Jason Potter, Kim Rosenthal
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Abstract
The invention relates to reverse transcriptases which have increased fidelity (or reduced misincorporation rate) and/or terminal deoxynucleotidyl transferase activity. In particular, the invention relates to a method of making such reverse transcriptases by modifying or mutating specified positions in the reverse transcriptases. The invention also relates to nucleic acid molecules containing the genes encoding the reverse trancriptases of the invention, to host cells containing such nucleic acid molecules and to methods to make the reverse trancriptases using the host cells. The reverse transcriptases of the invention are particularly suited for nucleic acid synthesis, sequencing, amplification and cDNA synthesis.
Claims22
A mutant MMLV reverse transcriptase comprising a polymerase domain having RNA-dependent DNA polymerase activity and a substitution in the amino acid sequence of the wild type MMLV polymerase domain within SEQ ID NO:
- 6, wherein amino acid number 1 of SEQ ID NO:
- 6 is the threonine following the initial methionine, and wherein said reverse transcriptase comprises at least one mutation at an amino acid position selected from the group consisting of Tyr64, Arg116, Lys152, Gln190, Thr197, and Phe309.
A mutant MMLV reverse transcriptase comprising a polymerase domain having a Val223 to His223 substitution in the amino acid sequence of the wild type MMLV polymerase domain within SEQ ID NO:
- 6, wherein amino acid number 1 of SEQ ID NO:
- 6 is the threonine following the initial methionine.
Dependencies are inferred from the claim text.