IP Library › Granted Patent US 10,612,009
Granted Patent B2
US 10,612,009 · App. 16/398,089 · Granted Apr 7, 2020

Polymerase variants

Inventors: Aruna Ayer (Santa Clara, CA); Cleoma Renetta Arnold (San Jose, CA); Arkadiusz Bibillo (Cupertino, CA); Mara Boenitz-Dulat (Tutzing, DE); Barbara Eckert (El Cerrito, CA); Ilya Lederman (San Francisco, CA); Colin Alexander McGaw (Santa Clara, CA); Charles Wayan Schwab (San Ramon, CA); Tyler O'Brien Shultz (Santa Clara, CA); Shawn Suko (El Sobrante, CA); Eileen Thai (San Jose, CA); Bigna Woersdoerfer (Basel, CH); David Daniel Wunderlich (Penzberg, DE)
Assignees: Roche Sequencing Solutions, Inc.; Roche Molecular Systems, Inc.
C12N9/1252C07K2319/21C12Y207/07007
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Quick Facts
Patent No.
US 10,612,009
App. No.
16/398,089
Granted
Apr 7, 2020
Kind
B2
Abstract

Described herein is a variant pol6 polymerase having at least one mutation selected from H223, N224, Y225, H227, I295, Y342, T343, I357, S360, L361, I363, S365Q, S366, Y367, P368, D417, E475, Y476, F478, K518, H527, T529, M531, N535, G539, P542, N545, Q546, A547, L549, I550, N552, G553, F558, A596, G603, A610, V615, Y622, C623, D624, I628, Y629, R632, N635, M641, A643, 1644, T647, I648, T651, I652, K655, W656, D657, V658, H660, F662, L690 and combinations thereof.

Claims (12)

1. An isolated polypeptide having a DNA polymerase activity comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1, wherein the amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1 comprises at least two amino acid substitutions relative to SEQ ID NO: 2, wherein one of said at least two amino acid substitutions corresponds to position T529 of SEQ ID NO: 2.

2. The isolated polypeptide of claim 1 , wherein the T529 substitution is a T529M/F substitution.

3. The isolated polypeptide of claim 1 , wherein said isolated polypeptide has a substitution corresponding to T529M.

4. The isolated polypeptide of claim 1 , wherein said at least two amino acid substitutions comprises a set of substitutions selected from the group consisting of:

T529M+T647G+A547F;

T529M+A610T+A547F;

M641Y+T529M+A547F;

N535L+N545K+T651Y+T529M; and

N535I+N545K+T651Y+T529M.

5. The isolated polypeptide of claim 1 , wherein said polymerase comprises less than 10 mutations.

6. The isolated polypeptide of claim 1 , wherein at least one of the at least two corresponds to a position of SEQ ID NO: 2 selected from the group consisting of selected from the group consisting of Y225, M531, N535, G539, N545, A547, N552, A610, M641, Y629, T647, T651, I652, K655, D657, V658.

7. The isolated polypeptide of claim 1 , wherein at least one of the at least two substitutions is selected from the group consisting of selected from the group consisting of Y225L/T/I/F/A, M531H/Y/A/K/R/W/T/L/V, N535L/Y/M/K/I, G539Y/F, N545K/D/S/L/R, A547M/Y/W/F/V/S, N552L/M/S, A610T/E, M641L/Y, Y629 W/H/M, T647G/A/E/K/S, T651Y/F/M, I652Q/G/S/N/F/T, K655G/F/E/N, D657R/P/A, and V658L.

Assignments (1)
MERGER AND CHANGE OF NAME Recorded Feb 25, 2020
From: GENIA TECHNOLOGIES, INC.; ROCHE SEQUENCING SOLUTIONS, INC.
To: ROCHE SEQUENCING SOLUTIONS, INC.
Reel/Frame 051918/0560 →
Continuity (5)
Division 15012317 · Feb 1, 2016
Provisional Application 62111034 · Feb 2, 2015
Provisional Application 62161571 · May 14, 2015
Provisional Application 62202895 · Aug 9, 2015
Related Publication 20190249154A1 · Aug 15, 2019