Granted Patent B2

US 12,060,587 · App. 17/226,693 · Granted Aug 13, 2024

Inventors: Zhenyu Zhu (Beverly, MA); Aine Quimby (Newton, NH); Shengxi Guan (Stoneham, MA); Dapeng Sun (Arlington, MA); Yishu Huang (Shanghai, CN); Xuhui Lai (Shanghai, CN); Siu-hong Chan (Ipswich, MA); Xianghui Li (Shanghai, CN); Shuang-Yong Xu (Lexington, MA); Chunhua Zhang (Ipswich, MA)

Assignee: New England Biolabs, Inc.

C12N9/22C12N9/16C12Q1/34C12Q1/44C12Q1/68C12Y301/21004G01N2333/916G01N2333/922

Abstract

Claims (4)

1. A composition comprising a variant KpnI restriction endonuclease having reduced star activity as compared to a wildtype KpnI restriction endonuclease comprising the amino acid sequence of SEQ ID NO: 4, wherein the variant KpnI restriction endonuclease comprises the amino acid sequence according to SEQ ID NO:4 except for having (a) an amino acid substitution at a position corresponding to position 16 of SEQ ID NO:4; or (b) an amino acid substitution at a position corresponding to position 148 of SEQ ID NO:4; or (c) amino acid substitutions at positions corresponding to positions 16 and 148 of SEQ ID NO:4.

2. The composition according to claim 1 , wherein the substitution at the position corresponding to position 16 of SEQ ID NO:4 is a D16N substitution.

3. The composition according to claim 1 , wherein the substitution at the position corresponding to position 148 of SEQ ID NO:4 is a D148E substitution.

4. The composition according to claim 1 , wherein the variant KpnI restriction endonuclease comprises the amino acid sequence according to SEQ ID NO:4 except for having substitutions D16N and D148E.

Assignments (2)

Continuity (10)