Invention Grant
US09273301B2 Thermostable blunt-end ligase and methods of use 有权
热稳定性连接酶及其使用方法

  • Patent Title: Thermostable blunt-end ligase and methods of use
  • Patent Title (中): 热稳定性连接酶及其使用方法
  • Application No.: US14214834
    Application Date: 2014-03-15
  • Publication No.: US09273301B2
    Publication Date: 2016-03-01
  • Inventor: Frederick Christians
  • Applicant: Theranos, Inc.
  • Applicant Address: US CA Palo Alto
  • Assignee: Theranos, Inc.
  • Current Assignee: Theranos, Inc.
  • Current Assignee Address: US CA Palo Alto
  • Main IPC: C12N9/00
  • IPC: C12N9/00 C12P19/34
Thermostable blunt-end ligase and methods of use
Abstract:
Fusion proteins having thermostable blunt-end ligase activity are provided. Blunt-end ligases are useful for DNA amplification, sequencing, production of recombinant DNA and recombinant fusion proteins, and other purposes. These thermostable blunt-end DNA ligases are useful in ligation schemes which include, e.g., an incubation at about 60-65° C. or higher, or as high as about 94° C., or at other temperatures. The ligases disclosed herein may enable high temperature blunt-end ligation without need for molecular crowding agents, and so may be useful for many nucleic acid ligation-amplification schemes, e.g., ones which operate at a uniform temperature (e.g., at about 60° C. or higher), including ones which require temperature cycling, e.g., from about 94° C. to about 60° C. (or higher) for one, two, three, or more cycles. The thermostable blunt-end DNA ligases disclosed herein enable higher specificity target amplification, for example, by permitting temperature denaturation of double-stranded DNA templates as well as specific primer binding.
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