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    THERMOLABILE PROTEINASES
    2.
    发明申请
    THERMOLABILE PROTEINASES 审中-公开

    公开(公告)号：WO2019170809A1

    公开(公告)日：2019-09-12

    申请号：PCT/EP2019/055724

    申请日：2019-03-07

    Applicant: ARCTICZYMES AS

    Inventor： STRIBERNY, Bernd Ketelsen ,  PEDERSEN, Cathrine ,  HENRIKSEN, Jørn Remi ,  LANES, Olav ,  LORENTZEN, Marit Sjo

    IPC: C12N9/52

    Abstract: The invention provides a composition comprising a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said composition is ≤ about 80 µ M; or ii) the concentration of monovalent salt in said composition is ≥ about 20 mM. Under such conditions, the proteinases and enzymatically active fragments thereof are inducibly thermolabile. The invention further provides samples comprising one or more polypeptides and a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said sample is ≤ about 80 µ M; or ii) the concentration of monovalent salt in said sample is ≥ about 20 mM. The invention further provides methods comprising the inactivation of such proteinases or enzymatically active fragments thereof, wherein said method comprises the step of heating the sample to inactivate said proteinase or enzymatically active fragment, and wherein i) the concentration of free calcium in said sample is ≤ about 80 µ M; or ii) the concentration of monovalent salt in said sample is ≥ about 20 mM.

    THERMOLABILE EXONUCLEASES
    3.
    发明申请
    THERMOLABILE EXONUCLEASES 审中-公开
    可燃烧剂

    公开(公告)号：WO2016026574A1

    公开(公告)日：2016-02-25

    申请号：PCT/EP2015/001703

    申请日：2015-08-19

    Applicant: ARCTICZYMES AS ,  UNIVERSITETET I TROMSØ - NORGES ARKTISKE UNIVERSITET

    Inventor： SOLSTAD, Terese ,  ANDREASSEN, Elisabeth, Lill ,  LORENTZEN, Marit, Sjo ,  LANES, Olav ,  ELDE, Morten ,  LARSEN, Atle, Noralf ,  PIOTROWSKI, Yvonne ,  WILLASSEN, Nils, Peder

    IPC: C12N9/22

    CPC classification number: C12Q1/6848 ,  C12N9/22 ,  C12Q1/34 ,  C12Q2521/325 ,  C12Y301/11001 ,  G01N1/30 ,  G01N2333/922

    Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID No. 1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55 °C for 10 minutes in a buffer consisting of 10 mM Tris-HCI, pH 8.5 at 25 °C, 50 mM KCI and 5 mM MgCI 2 ; (ii) is substantially specific for single stranded DNA; and (iii) has a 3'-5' exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used. The invention still further provides nucleic acids encoding said exonuclease or an enzymatically active fragment thereof and kits or compositions comprising the same.

    Abstract translation: 本发明提供了外切核酸酶或其酶促活性片段，所述外切核酸酶具有SEQ ID No.1的氨基酸序列或与其至少约50％相同的氨基酸序列，其中所述外切核酸酶或酶活性片段（i ）通过在25℃，50mM KCl和5mM MgCl 2的10mM Tris-HCl，pH8.5组成的缓冲液中在约55℃的温度下加热10分钟而基本上不可逆地灭活。 （ii）对单链DNA基本上是特异性的; 和（iii）具有3'-5'核酸外切酶活性。 本发明还提供从样品中除去单链DNA的方法，核酸扩增方法，逆转录方法以及使用其核酸外切酶或酶促活性片段的核酸序列分析方法。 本发明还提供了编码所述外切核酸酶或其酶促活性片段的核酸和包含其的试剂盒或组合物。

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