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    THERMOLABILE EXONUCLEASES
    4.
    发明公开
    THERMOLABILE EXONUCLEASES 审中-公开
    热可塑性外切变

    公开(公告)号：EP3183340A1

    公开(公告)日：2017-06-28

    申请号：EP15770444.6

    申请日：2015-08-19

    申请人： Arcticzymes AS ,  Universitetet I Tromsø - Norges Arktiske Universitet

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

    IPC分类号： C12N9/22

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

    摘要： 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 MgCI2; (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.

    摘要翻译： 本发明提供了核酸外切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％同一性的氨基酸序列，其中所述外切核酸酶或其酶促活性片段（i ）通过在由pH为8.5的10mM Tris-HCl，25℃，50mM KCl和5mM MgCl 2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失活; （ii）对单链DNA基本上是特异性的; 和（iii）具有3'-5'外切核酸酶活性。 本发明进一步提供了从样品中去除单链DNA的方法，核酸扩增方法，逆转录方法和核酸序列分析方法，其中使用了核酸外切酶或其酶活性片段。 本发明还进一步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核酸酶或组合物。