公开(公告)号：US09127269B2

公开(公告)日：2015-09-08

申请号：US14028151

申请日：2013-09-16

Applicant: Cornell Research Foundation, Inc.

Inventor： Francis Barany ,  Weiguo Cao ,  Jie Tong

IPC: C12N9/00

CPC classification number: C12N9/93

Abstract: The present invention is directed to a mutant thermostable ligase having substantially higher fidelity than either T4 ligase or Thermus thermophilus ligase. The ligase of the present invention is a mutant of a wild-type thermostable ligase having a histidine adjacent a KXDG motif, where the mutant thermostable ligase has a mutation in its amino sequence where the histidine adjacent the KXDG motif in the wild-type thermostable ligase is replaced with an arginine, and wherein X is any amino acid. The DNA molecule encoding this enzyme as well as expression systems and host cells containing it are also disclosed. The thermostable ligase of the present invention is useful in carrying out a ligase detection reaction process and a ligase chain reaction process.

Abstract translation: 本发明涉及一种具有比T4连接酶或Thermus thermophilus连接酶具有更高保真度的突变热稳定连接酶。 本发明的连接酶是具有与KXDG基序相邻的组氨酸的野生型热稳定连接酶的突变体，其中突变热稳定连接酶在其氨基序列中具有突变，其中野生型热稳定连接酶中KXDG基序相邻的组氨酸 被精氨酸代替，其中X是任何氨基酸。 还公开了编码该酶的DNA分子以及含有它的表达系统和宿主细胞。 本发明的热稳定连接酶可用于进行连接酶检测反应过程和连接酶链反应过程。