Polymerase Variants and Uses Thereof
US-2016333327-A1 · Nov 17, 2016 · US
Polymerase variants
US10308918B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-10308918-B2 |
| Application number | US-201615012317-A |
| Country | US |
| Kind code | B2 |
| Filing date | Feb 1, 2016 |
| Priority date | Feb 2, 2015 |
| Publication date | Jun 4, 2019 |
| Grant date | Jun 4, 2019 |
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Abstract
Official abstract text for this publication.
Described herein are variant pol6 polymerase having at least having at least two substitutions, wherein at least one of the substitutions is at position S366 of SEQ ID NO: 2. The disclosed polymerases may be coupled to alpha-hemolysin nanopores, and may be used in nanopore-based sequencing apparatuses.
First claim
Opening claim text (preview).
What is claimed is: 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 S366 of SEQ ID NO: 2 and wherein said amino acid corresponding to position S366 is substituted with A or L. 2. The isolated polypeptide of claim 1 , wherein said at least two amino acid substitutions comprises a set of amino acid substitutions selected from the group consisting of: S366A+N535L; S366L+G553S+F558P; S366A+N535L+A547M; S366A+P542E+N545K; S366A+P542E+I652Q; S366A+N535L+T529M; S366A+N535L+I652Q; S366A+N535L+N545K; S366A+N535I+I652Q; T651Y+S366A+A547F; A547F+A610T+S366A; S366A+T647G+A547F; T529M+S366A+A547F; T647E+S366A+A547F; N545K+S366A+A547F; S366L+F478L+A596S+L690M; S366A+N535L+I652Q+T529M; S366A+S365A+P368G+G603T; S366A+N535L+I652Q+A547Y; S366A+N535L+A547M+T647G; T529M+S366A+A547F+N545K; T529M+S366A+A547F+N545R; T529M+S366A+A547F+N552L; T529M+S366A+A547F+Y629W; S366A+N535L+A547M+T647G+S360G; S366A+N535L+I652Q+A547Y+K655G; T529M+S366A+A547F+N545L+Y629W; T529M+S366A+A547F+N545L+Y225L; T529M+S366A+A547F+N545L+Y225F; T529M+S366A+A547F+N545L+K655F; T529M+S366A+A547F+N545L+N552L; T529M+S366A+A547F+N545R+M531A; T529M+S366A+A547F+N545R+G539Y; T529M+S366A+A547F+N545R+V658L; T529M+S366A+A547F+N545L+Y225L+D657R; T529M+S366A+A547F+N545L+Y225L+N552L; T529M+S366A+A547F+N545L+Y225L+I652G; T529M+S366A+A547F+N545L+Y225L+I652Q; and T529M+S366A+A547F+N545L+Y225L+N552M. 3. The isolated polypeptide of claim 1 comprising S366A+T529M+N545L+A547F. 4. The isolated polypeptide of claim 3 further comprising at least one substitution selected from the group consisting of: a. Y225L/F/A; b. M531A; c. G539Y; d. N552L; e. Y629W; and f. K655F. 5. The isolated polypeptide of claim 1 comprising S365A+S366A+P368G+G603T. 6. The isolated polypeptide of claim 1 , comprising T529M+S366A+A547F. 7. The isolated polypeptide of claim 1 , comprising T529M+S366A+A547F+N545L+Y225L. 8. The isolated polypeptide of claim 1 , comprising T529M+S366A+A547F+N545L+Y225L+D657R. 9. The isolated polypeptide of claim 1 , wherein said at least two amino acid substitutions further comprises one or more substitutions selected from the group consisting of: F478L, T529M, N535L, N545K/L, A547F, G553S, T647G, and I652Q.
Assignees
Inventors
Classifications
- C12N9/1252Primary
DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase · CPC title
DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase · CPC title
containing a His-tag · CPC title
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- What does patent US10308918B2 cover?
- Described herein are variant pol6 polymerase having at least having at least two substitutions, wherein at least one of the substitutions is at position S366 of SEQ ID NO: 2. The disclosed polymerases may be coupled to alpha-hemolysin nanopores, and may be used in nanopore-based sequencing apparatuses.
- Who is the assignee on this patent?
- Genia Tech Inc, Roche Molecular Systems Inc
- What technology area does this patent fall under?
- Primary CPC classification C12N9/1252. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Jun 04 2019 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 5 related publications on this page (citations in our corpus or others sharing the same primary CPC).