High fidelity reverse transcriptases and uses thereof

Details High fidelity reverse transcriptases and uses thereof High fidelity reverse transcriptases and uses thereof

C - Chemistry, Metallurgy

12

N

C12N 15/54 (2006.01) C07H 21/04 (2006.01) C12N 1/21 (2006.01) C12N 5/00 (2006.01) C12N 9/00 (2006.01) C12N 9/12 (2006.01) C12N 15/10 (2006.01) C12N 15/63 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2401417

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 transcriptases of the invention, to host cells containing such nucleic acid molecules and to methods to make the reverse transcriptases using the host cells. The reverse transcriptases of the invention are particularly suited for nucleic acid synthesis, sequencing, amplification and cDNA synthesis.

L'invention se rapporte à des transcriptases inverses qui possèdent une fidélité accrue (ou un taux de mésincorporation réduit) et/ou une activité de désoxynucléotidyle transférase terminale. L'invention se rapporte notamment à un procédé de fabrication de ces transcriptases inverses par modification ou mutation de positions spécifiées dans lesdites transcriptases inverses. L'invention se rapporte également à des molécules d'acides nucléiques contenant les gènes codant lesdites transcriptases inverses, à des cellules hôtes contenant de telles molécules d'acides nucléiques et aux procédés de fabrication de ces transcriptases inverses au moyen desdites cellules hôtes. Les transcriptases inverses de la présente invention sont particulièrement adaptées à la synthèse d'acides nucléiques, à la détermination des séquences nucléotidiques, à l'amplification et à la synthèse d'ADNc.

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