CN101287828A

CN101287828A - 嗜热球菌属某种(菌株9°n-7)dna连接酶的发现、克隆和纯化

Info

Publication number: CN101287828A
Application number: CNA2006800383873A
Authority: CN
Inventors: I·希尔德克劳特; E·希尔德克劳特
Original assignee: New England Biolabs Inc
Current assignee: New England Biolabs Inc
Priority date: 2005-09-15
Filing date: 2006-09-15
Publication date: 2008-10-15
Also published as: WO2007035439A8; JP2009508488A; US20090142811A1; EP1929000A1; WO2007035439A1

Abstract

描述了从嗜热球菌属某种(Thermococcus sp.)(菌株9°N－7)分离的热稳定的DNA连接酶的组合物，以及制备和使用该组合物的方法。在连接过程中，热稳定的DNA连接酶依赖于ATP而不是NAD⁺作为辅因子，并且在100℃下，保持活性。

Description

嗜热球菌属某种(菌株9°N-7)DNA连接酶的发现、克隆和纯化

技术背景

[001]嗜热球菌属(Thermococcus)是古细菌门(Archaea)的一个属。这些古老的生物在包括高温的极端条件下的多种环境中生长。在实验室中培养这些生物的能力是非常有限的，以致关于它们的生物化学或它们的代谢鲜为人知。

[002]连接酶是在双链体DNA中的单链切口位点上催化磷酸二酯键形成的酶。连接酶也催化双链体DNA的共价键，一般是平末端对平末端，或者一个粘末端对另一个粘末端。连接酶已经从多种细菌中克隆，包括一种热稳定连接酶——水生栖热菌(Thermus aquaticus)(Taq连接酶)。该连接酶已在美国专利第6,054,564号中描述。

[003]仅一些嗜热球菌已被分离，关于它们可能含有的连接酶的特性或者编码这类蛋白的基因知之甚少(参见，例如Nakatani等，J.Bacteriology 182：6424-6433(2000))。从不同属的古细菌——热球菌属(pyrococcus)——得到的连接酶例如在美国专利第5,506,137和5,700,672号以及在Keppetipola等J.Bacteriology 187：6902-6908(2005)中已被描述。

[004]连接酶已被用于分子生物学的许多技术中，包括DNA扩增、测序和单核苷酸多态性的检测。寻找在高温下稳定并且具有快速动力学和严格特异性的改善的连接酶一直是所需求的。

发明概述

[005]在本发明的一实施方式中，提供了具有DNA连接酶活性的基本上纯的重组蛋白质，其中该蛋白质与SEQ ID NO：13具有至少91％的氨基酸序列同一性。

[006]在本发明的进一步实施方式中，提供具有DNA连接酶活性的基本上纯的蛋白质，其中DNA连接酶用选自下列的DNA序列编码：(a)与SEQ ID NO：2基本上相同的序列；(b)与SEQ ID NO：2基本上互补的序列；(c)在严格条件下，与SEQ ID NO：2基本上杂交的序列；和(d)编码SEQ ID NO：13的序列。

[007]在上述实施方式中指出的蛋白质的进一步的特征在于在大约100℃的温度下温育30分钟后，连接酶保留了至少25％的连接酶活性。而且，该连接酶可能的进一步特征在于在连接过程中其使用ATP作为辅因子，然而就这一目的而言，NAD⁺没有提供可检测的效用。

[008]在进一步的实施方式中，提供了编码DNA连接酶的DNA，该DNA具有选自下列的序列：(a)与SEQ ID NO：2基本上相同的序列；(b)与SEQ ID NO：2基本上互补的序列；(c)在严格条件下，与SEQ IDNO：2基本上杂交的序列；和(d)编码SEQ ID NO：13的序列。

[009]在进一步的实施方式中，描述了含有上述DNA的载体。另外，提供了能从载体表达连接酶的宿主细胞。

[010]在进一步的实施方式中，提供了连接磷酸二酯键的方法，该方法包括：选择上述类型的DNA连接酶；将连接酶与DNA混合，所述DNA在该DNA的至少一条链上含有切口；并且在所述切口连接所述磷酸二酯键。

[011]在该方法的一实例中，DNA连接酶是从古细菌分离物，具体为嗜热球菌属某种(Thermococcus sp.)(菌株9°N-7)得到的热稳定连接酶。

附图简述

[012]图1a-1至1a-5示出9°N-7DNA连接酶变体(SEQ ID NOS：1-7)的DNA序列比对。

[013]图1b-1至1b-2示出嗜热球菌属某种(菌株9°N-7)DNA连接酶变体(SEQ ID NOS：8-15)的蛋白质比对。

[014]图2示出***到litmus 28i内的嗜热球菌属某种(菌株9°N-7)DNA连接酶基因的质粒图。

[015]图3示出***到pMalC2x内的嗜热球菌属某种(菌株9°N-7)DNA连接酶基因的质粒图。

[016]图4示出嗜热球菌属某种(菌株9°N-7)(SEQ ID NO：15)和喜烟嗜热球菌(Thermococcus fumicolans)(SEQ ID NO：16)、克达卡拉嗜热球菌(Thermococcus kodakaraensis)(SEQ ID NO：17)、深海热球菌(Pyrococcus abyssi)(SEQ ID NO：18)和激烈热球菌(pyrococcusfurious)(SEQ ID NO：19)的蛋白质比对。

[017]图5示出磷酸纤维素柱组份(phosphocellulose column fraction)的SDS PAGE。泳道如下标记：

FT(柱流通)指组份编号23-34；

MW指分子量标准。

箭头指出凝胶上与DNA连接酶相应的带的位置。

[018]图6示出9°N-7DNA连接酶的热稳定性。30μl、pH7.5的10mM Tris HCl、2.5mM MgCl₂、2.5mM DTT、300μM ATP和0.1％TritonX-100——其含有3μl以1∶100稀释的纯9°N-7DNA连接酶，在pH7.5的10mM Tris HCl、2.5mM MgCl₂，、2.5mM DTT、300μM ATP和0.1％Triton X-100进一步连续三倍稀释。四组相同的稀释液在4℃、80℃、90℃或100℃下温育30分钟。

[019]为了终止反应，将样品置于冰上并且将等体积的pH7.5的10mM Tris HCl、2.5mM MgCl₂，、2.5mM DTT、300μM ATP、0.1％TritonX-100和50μg/ml BstEIIλDNA加入到每支管中。然后在45℃下温育反应物15分钟，在此之后将0.15体积的50％的甘油、100mM EDTA和溴酚兰加入到每支管中。然后在75℃下温育反应物5分钟，并且在1％琼脂糖TBE凝胶上进行电泳。

[020]图A示出在冰上温育30分钟的结果。

[021]图B示出在80℃下温育30分钟的结果。

[022]图C示出在90℃下温育30分钟的结果。

[023]图D示出在100℃下温育30分钟的结果。

[024]对于每一个图，泳道如下指定：

泳道1表示没有进一步稀释；

泳道2以1∶3稀释；

泳道3以1∶9稀释；

泳道4以1∶27稀释；和

泳道5以1∶81稀释。

[025]图7示出的凝胶中，9°N-7聚合酶与Taq聚合酶在含有大肠杆菌(E.coli)聚合酶和大肠杆菌内切酶IV(Endo IV)的修复混合物中比较。该修复混合物与脱嘌呤的DNA一起温育，并且进行扩增。

泳道1是对照。

泳道2是在没有修复混合物下的DNA；

泳道3和4是DNA和含有480单位Taq连接酶的修复混合物的复制样品；和

泳道5和6是DNA和含有500单位9°N-7连接酶的修复混合物的复制样品。

实施方式详述

[026]这里使用的术语“热稳定连接酶(thermostable ligase)”指催化DNA连接和在100℃下、30分钟后保持其至少25％活性的酶。在极端温度下的热稳定性是将嗜热球菌属连接酶(古细菌)和嗜热菌属(Thermus)连接酶(细菌)区分开的特征。

[027]嗜热球菌属某种(菌株9°N-7)是从水热喷口(hydrothermalvent)分离的嗜热球菌的一个种(Southworth等PNAS 93：5281(1996))。

[028]嗜热球菌属某种(菌株9°N-7)DNA连接酶的两个已知的最接近的亲属(relative)是喜烟嗜热球菌连接酶和克达卡拉嗜热球菌连接酶(JP 2000308494-A/l)，其在氨基酸水平分别具有88％和90％的同一性。这两个连接酶都被报道利用NAD⁺或ATP作为辅因子，从而构成一组新类型连接酶。喜烟嗜热球菌连接酶被Nakatani等(J.Bacteriology182：6424-6433(2000))报道利用NAD⁺或ATP同样好，并且克达卡拉嗜热球菌连接酶在利用NAD⁺代替ATP时具有减低水平的活性，然而嗜热球菌属某种(菌株9°N-7)利用NAD⁺时没有可检测的活性(实施例2)。

[029]术语“基本上相同”和“基本上互补”拟指DNA或氨基酸序列与鉴定的序列很大的相同性或同一性，或者与互补序列很大的相同性或同一性。该术语拟包括在氨基酸或DNA序列中含有与在图中规定的最小差异的序列。这种差异可由不明显干扰蛋白质连接功能的诱变事件引起。

[030]在本发明的一实施方式中，在下列条件下进行严格杂交：a)杂交：0.75M NaCl、0.15Tris HCl、10mM EDTA、0.1％NaCl、0.1％SLS、0.03％BSA、0.03％Ficoil 400、0.03％PVP和100μg/ml煮沸的小牛胸腺DNA，在50℃下大约12小时，和；b)用0.1X SET、0.1％SDS、0.1％NaCl和0.1M磷酸盐缓冲液，在45℃下洗涤3次共30分钟，并在DNA印迹(Southern Blot)上检测双链杂交的DNA的存在。

[031]本文引用的所有参考文献以及2005年9月15号提交的美国临时专利申请第60/717,296号被引入作为参考。

实施例

实施例I：使用简并引物克隆嗜热球菌属某种(菌株9°N-7)DNA连接酶基因

[032]首先通过PCR从嗜热球菌属某种(菌株9°N-7)的基因组DNA扩增基因。从Nakatani等J.Bact.182(22)：6424-6433(2000)(克达卡拉嗜热球菌)和Rolland等FEMS Microbiology Lett.236(2)：267-273(2004)(喜烟嗜热球菌)的参考文献获得正向引物序列。具有指定简并性的共有序列设计如下：

正向引物

5′CGGTGGTGCATATGRGCGAYATGMRSTACTC(SEQ ID NO：20)

反向引物

5′ATAAACTCTAGATTACYTCTTCGCCTTGAACCTCTCCTGG(SEQ ID NO：21)

[033]使用嗜热球菌属某种(菌株9°N-7)的引物从嗜热球菌属某种(菌株9°N-7)的基因组DNA扩增DNA连接酶的基因。用来克隆基因的PCR反应条件如下：

[034]100μl反应混合物——含有pH 8.8的20mM Tris-HCL、10mM KCl、10mM(NH₄)₂SO₄和4mM MgSO₄、0.1％Triton X-100、200μM的每种dNTP、50ng嗜热球菌属某种(菌株9°N-7)的基因组DNA、正向和反向引物每种500ng、2.5单位的TaqDNA聚合酶和0.02单位的

DNA聚合酶，被加热到94℃1分钟，然后在45℃1分钟，并且然后在72℃3分钟。重复30次该温度循环。在完成循环后，将反应温度降低至室温，并且加入5单位大肠杆菌DNA聚合酶Klenow片段，并在室温下进一步温育5分钟。然后，将反应物调整为70mM EDTA。PCR产物用苯酚提取、用醇沉淀，并在CL6B琼脂糖自旋柱(sepharose spincolumn)上对其脱盐。

[035]将1700bp PCR产物克隆到大肠杆菌中。EcoRV-切割的litmus 28i被用作克隆DNA片段的载体。

[036]T4DNA连接酶缓冲液中的10μl连接反应物包含80ng***片段、80ng litmus载体和400单位T4DNA连接酶(New England Biolabs，Inc.，Ipswich，MA)。连接反应在16℃温育过夜，经电穿孔进入大肠杆菌TB1细胞，并在IPTG XGAL平板上铺板。

[037]挑选白色的菌落。9个白色菌落中的一个具有1700bp***片段。独立的电穿孔产生具有1700bp***片段的另一个克隆。对这两个克隆中的***片段进行测序。

[038]从这些克隆的序列，新的具有较少简并性的正向引物设计如下：

9°N正向引物：

5′cggtggtgcatatgggcgayatgaggtactccgagctgg(SEQ ID NO：22)

(2)使用比在实施例(1)中的引物具有较少简并性的第二正向引物克隆嗜热球菌属某种(菌株9°N-7)的连接酶

[039]使用9°N-7正向引物实施另外4个独立的PCR反应，其仅包含一个简并碱基代替了上述(1)中的正向引物——其含有5个简并碱基。

[040]100μl含有50ng嗜热球菌属某种(菌株9°N-7)基因组DNA、正向和反向引物每种500ng、200μM的每种dNTP和1μl Phusion DNA聚合酶(New England Biolabs，Inc，Ipswich，MA)的Phusion HF缓冲液(New England Biolabs，Inc，Ipswich，MA)被加热到98℃持续30秒，然后作25个下列过程的循环：98℃10秒，然后70℃30秒，接下来72℃1分钟。然后在72℃温育反应物5分钟。每一PCR反应的产物作为初始PCR反应物处理，并被克隆到litmus 28i中，如上所述。通过小量制备DNA，从PCR反应得到的两个独立克隆(A1和A3)被证实含有1700碱基对***片段，以及从其它三个PCR反应(B2、C3、D3)的每一个得到的一个克隆同样被证实含有1700碱基对***片段。然后培养这些克隆，并将它们的粗提物在SDS PAGE上电泳。每一克隆表达60kd的蛋白质。

[041]从克隆A1、A3、B2、C3、D3得到的质粒和另外的lig7和lig8被纯化，并且对***片段测序。在图1a-1到1a-5中提供了DNA序列(SEQ ID NOS：1-7)。

[042]尽管不希望被理论所限制，但是在序列中观察到的小差异可由嗜热球菌属某种(菌株9°N-7)细胞群内的克隆变异解释。序列变异是所有第三位置变化，或者保守氨基酸变化。克隆B2是连接酶共有序列的代表。DNA连接酶首次在紧密控制的表达载体中表达(图2)。

(3)在大肠杆菌中表达连接酶基因(B2)

[043]通过用NdeI和XbaI切割，从litmus载体上剪切B2片段。从琼脂糖凝胶上切下1700bp片段，并且用琼脂糖酶消化凝胶切片以释放所述片段。

[044]通过用NdeI和XbaI切割制备表达载体pMalC2X(NewEngland Biolabs，Inc.，Ipswich，MA)，并且使其去磷酸化。在10μl含有400ng***片段和100ng T4DNA连接酶缓冲液中的载体以及200单位T4DNA连接酶的反应物中，在16℃温育16小时，将切割的1700bp碱基对PCR片段连接到pMalC2X载体。将连接反应物电穿孔入大肠杆菌TB1细胞，并且携带1700bp片段的克隆被分离并且命名为嗜热球菌属某种(菌株9°N-7)B2-1(图3)。

[045]在LB培养基中生长克隆，并且用IPTG进行诱导。诱导的细胞的样品被裂解，并且在SDS PAGE凝胶上电泳，以显示出与约60kd大小的蛋白质相应的带。对从DNA序列得到的蛋白质序列的分析表明基因编码带有26个罕用精氨酸密码子的蛋白质。因此，含有用于精氨酸的罕用tRNA的宿主细胞(大肠杆菌BL-2(DE3)RIL)(Stratagene，LaJoIIa，CA)被用来获得更高水平的表达。在嗜热球菌属某种(菌株9°N-7)诱导后，宿主样品中的B2-1质粒通过SDS PAGE进行分析，并且观察到明显的60kd带。

(4)嗜热球菌属某种(菌株9°N-7)连接酶与其它热稳定DNA连接酶的比较

[046]通过CLUSTAL多序列比对，将嗜热球菌属某种(菌株9°N-7)DNA连接酶氨基酸序列与其它4种嗜热DNA连接酶进行比较。CLUSTAL W(1.82)多序列比对(Multiple Sequence Alignments)序列形式为Pearson。

序列1：9°N-7-B2(SEQ ID NO：15) 564aa

序列2：克达卡拉嗜热球菌(SEQ ID NO：16) 562aa

序列3：深海热球菌(SEQ ID NO：17) 559aa

序列4：激烈热球菌(SEQ ID NO：18) 561aa

序列5：喜烟嗜热球菌(SEQ ID NO：19) 559aa

同一性得分

序列(1∶2)比对得分：90

序列(1∶3)比对得分：81

序列(1∶4)比对得分：78

序列(1∶5)比对得分：88

序列(2∶3)比对得分：80

序列(2∶4)比对得分：80

序列(2∶5)比对得分：87

序列(3∶4)比对得分：90

序列(3∶5)比对得分：78

序列(4∶5)比对得分：77

[048]图4中给出比对。嗜热球菌属某种(菌株9°N-7)DNA连接酶已知最接近的亲属是克达卡拉嗜热球菌DNA连接酶，其中具有90％的氨基酸同一性和80.9％的核苷酸同一性。

(5)嗜热球菌属某种(菌株9°N-7)DNA连接酶的纯化

[049]使用含有从嗜热球菌属某种(菌株9°N-7)得到的DNA连接酶的B2片段的pMalC2X质粒(New England Biolabs，Inc.，Ipswich，MA)转化大肠杆菌BL-21(DE3)-RIL(Stratagene，La Jolla，CA)。在含有50μg/ml的氨苄青霉素和25μg/ml的氯霉素的100ml LB培养基中，在37℃下，生长细胞。在温育过夜后，将培养物转移到10升发酵罐中，并且在37℃下温育，直到达到0.59的OD600，并且加入0.1克的IPTG。温育培养物另外的5.75小时，并且收获。在-20℃，贮藏该细胞团(cell paste)。

[050]在40ml pH7.5的10mM Tris HCl、20mM NaCl、0.1mMEDTA和1.0mM DTT中的10克细胞团被解冻并通过超声波裂解。将提取物置入0.3mM PMSF和200mM NaCl。通过离心净化提取物。将净化的提取物通过0.2M NaCl中的DEAE琼脂糖柱。然后收集流过该柱的蛋白质，并且将其稀释为100mM的NaCl。将其施用到磷酸纤维素柱，并且用100mM到1.1M的NaCl梯度洗脱被吸收的蛋白质。通过SDS PAGE分析该级分(图5)，收集60kd的主峰，并且将其加热到75℃，30分钟。通过离心净化该溶液，将净化的溶液稀释为100mMNaCl，并且将其施用到羟磷灰石柱。将0-13％梯度的硫酸铵运用到该柱，收集级分，并且通过在含有50μg/ml的HindIII λDNA作为底物的T4DNA连接酶缓冲液(New England Biolabs，Inc.，Ipswich，MA)中温育多个级分，分析这些级分的活性。在37℃下，温育该反应物10分钟。通过加入10％的100mM EDTA和50％的甘油和溴酚兰染料终止反应。将反引物加热到65℃，并且将其负载到1％的琼脂糖凝胶上进行分析。

[051]收集含有大约80％连接酶活性的管，并将其在50％甘油、pH7.5的10mM Tris HCl、50mM KCl、10mM(NH₄)₂SO₄、0.1mM EDTA和1.0mM DTT中透析。纯化的嗜热球菌属某种(菌株9°N-7)DNA连接酶被贮藏在-20℃。

实施例2：嗜热球菌属某种(菌株9°N-7)DNA连接酶的特性

[052]推荐的反应条件是：

10mM Tris-HCl，pH7.5

2.5mM MgCl₂

2.5mM DTT

300μMATP

[053]在45℃下，用于分析活性的典型底物是λDNA。适当消化的λDNA可以在λDNA末端示出12-碱基延伸的连接状态。一般地，我们使用HindIII或BstEII预消化的λDNA。在琼脂糖凝胶电泳上实施连接分析。

[054]ATP的Km显示在100μM以下。利用Triton X-100刺激该活性。

[055]与喜烟嗜热球菌DNA连接酶不同，嗜热球菌属某种(菌株9°N-7)连接酶在存在NAD⁺的情况下，没有可检测的活性。

[056]该酶需要镁离子。2.5mM MgCl₂获得比10mM MgCl₂多10倍的活性。

[057]在大约100℃下温育该酶30分钟后，保留25％至50％之间的活性(图6)。

[058]在90℃下，连接酶能封闭带切口的DNA。DNA连接酶与BstNBI带切口的pUC19质粒DNA一起温育，并将松弛的带切口质粒转化成共价闭环DNA，如通过琼脂糖凝胶电泳所确定。在80℃时反应速率比在45℃时反应速率高。尽管带切口的质粒在90℃经历变性，但在90℃在变性前发生大量的切口封闭，这将所有带切口的质粒转变为单链质粒。

实施例3：嗜热球菌属某种(菌株9°N-7)DNA连接酶在DNA修复混合物中的应用

[059]使用包括菌株9°N-7DNA连接酶的酶的混合物，修复脱嘌呤所破坏的DNA。

[060]在实验反应中DNA通过Ide，H等Biochemistry 32(32)：8276-83(1993)描述的脱嘌呤而受损。λDNA(NEB#N3011，New EnglandBiolabs，Inc.，Ipswich，MA)被醇沉淀。将DNA以0.5mg/ml的浓度重悬浮在脱嘌呤缓冲液(100mM NaCl、10mM柠檬酸盐，pH 5.0)中，并且在70℃下温育120分钟。然后将样品醇沉淀，并且重新悬浮在0.01MTris、0.001M EDTA，pH 8.0的溶液中。在用缓冲液对照校准后，通过测量含DNA溶液的A²⁶⁰确定DNA的浓度。

[061]在下列酶的混合物中，在室温下温育受损的DNA达10分钟：DNA(1ng)；100μM dNTPs(NEB#M0447，New England Biolabs，Ipswich，MA)；1mM ATP；480单位Taq连接酶(NEB#M0208，New EnglandBiolabs，Ipswich，MA)或者500单位9°N-7DNA连接酶(NEB#M0238，New England Biolabs，Ipswich，MA)；0.1单位大肠杆菌DNA聚合酶I(大肠杆菌polI)(NEB#M0209，New England Biolabs，Inc.，Ipswich，MA)；10单位大肠杆菌内切酶IV(NEB#M0304，New England Biolabs，Inc.，Ipswich，MA)；1×Thermopol缓冲液(NEB#B9004，New England Biolabs，Inc.，Ipswich，MA)，至最终体积47.5μL。

[062]在反应结尾，将样品转移到冰上，然后扩增。阴性对照如上处理，但是没有使用酶。

DNA扩增反应

[063]根据Wang等Nucl.Acids Res.32：1197-1207(2004)的方法，使用下列引物，实施λDNA 的扩增：CGAACGTCGCGCAGAGAAACAGG(L72-5R)(SEQ ID NO：23)和CCTGCTCTGCCGCTTCACGC(L30350F)(SEQ ID NO：24)。

[064]将2.5μl的扩增混合物加入到47.5的上述修复混合物中。该扩增混合物含有100μM dNTPs、5单位Taq DNA聚合酶(New EnglandBiolabs，Inc.，Ipswich，MA)、0.1单位Vent

(exo+)DNA聚合酶(NewEngland Biolabs，Inc.，Ipswich，MA)、5×10^-11M引物L72-5R和5×10^-11M引物L30350F，于1×Thermopol缓冲液中。

[065]为了矫正酶储存缓冲液的影响，当修复酶从反应中省略时，将适当体积的储存缓冲液加入到反应物中。在所有的情况下，使用下列参数在热循环仪(thermal cycler)中进行扩增反应：在95℃下20秒1个循环，接下来在94℃下5秒，然后在72℃5分钟25个循环。被扩增的扩增子的大小为5kb。

[066]通过1％琼脂糖凝胶电泳，确定DNA(5kb)扩增的结果。将6×负载染料(Molecular Cloning：A Laboratory Manual，3rd ed.，eds.Sambrook and Russell，Cold Spring Harbor Press，Cold Spring Harbor，NY，2001，pp.5.4-5.17)加入到50μl扩增反应物中。然后，将20μl该溶液和作为尺寸标准的1μg的2-对数梯度分子量标准(2-logladder)(NEB#N3200，New England Biolabs，Inc.，Ipswich，MA)一起加载到琼脂糖凝胶上。

序列表

<110>新英格兰生物实验室公司

I.希尔德克劳特

E.希尔德克劳特

<120>嗜热球菌属某种(菌株9°N-7)DNA连接酶的发现、克隆和纯化

<130>NEB-268-PCN

<150>60/717,296

<151>2005-09-15

<150>PCT/US06/35919

<151>2006-09-15

<160>24

<170>PatentIn version 3.4

<210>1

<211>1695

<212>DNA

<213>未知

<220>

<223>嗜热球菌属某种(Thermococcus sp.)菌株9°N

<400>1

atgggcgata tgaggtactc cgagctggcc gaactctacc ggaggcttga gaagaccacg 60

ctcaaaacgc tcaagaccaa gttcgtcgcg gacttcctca agaaaacacc cgacgatttg 120

ctcgagatag ttccctacct gattctcggc aaggtctttc cggactggga cgagcgcgag 180

ctcggcgtcg gcgagaagct tctcataagg gccgtttcga tggcaaccgg cgtccccgag 240

agggaaatcg agaactcgat taaggacacc ggcgatttgg gcgagagcgt ggctctggct 300

ctaaagaaga ggaagcagaa gagcttcttc agccagcccc tcacgataaa gcgcgtctac 360

agcaccttcg ttaaggttgc cgaggcgagc ggagagggga gccaggacag gaagatgaag 420

tacttagcaa acctcttcat ggatgctcaa cccgaggagg gcaagtacat agccagaacc 480

gtcctcggga cgatgaggac gggcgtcgcc gagggaatcc tgcgcgatgc catagcggag 540

tccttcaagg tgaagccaga gctcgtcgag agggcctaca tgctcacgag cgacttcggc 600

tacgtggcga aggtcgccaa gctcgagggg aacgaggggc tctcgaaggt cagcatacag 660

attgggaagc cgataaggcc gatgctagct caaaacgccg ccaacgtcaa ggaagcgcta 720

atcgagatgg gcggtgaggc ggccttcgag attaagtacg acggcgcgcg cgttcaggtc 780

caccgcgatg gggacagggt gataatctac tcgaggaggc ttgagaacgt cacccgctcg 840

attcctgaga tagttgaggc ggtaaaggcc tccctgaagc cctctaatgt catagtcgag 900

ggcgagctgg ttgccgtcgg cgagaacggt cgcccgaggc ccttccagta cgtcctgagg 960

cgctttagga ggaagtacaa catcgaggag atgattgaga agattccgct cgagctcaac 1020

ctcttcgaca tcctctacgt tgaaggagag agcctcatcg acacgaagtt cgcagagagg 1080

aggaagaagc tcgaggagag cgttgaggag agcgataaga taaagctcgc cgaacagctc 1140

gttacgaaga aggttgaaga ggccgaggag ttctacaaga gggcccttga gctcggccac 1200

gagggcctga tggcgaagag gctggactcc atctacgagc ccggaaaccg cggtaagaag 1260

tggctgaaga ttaagcccac gatggagaac cttgacctcg tcattatcgg agccgaatgg 1320

ggcgagggca ggcgcgcgca cctgctcggc tcgttcctcg ttgcggccta cgaccccgag 1380

agcggtgagt tcgtcccggt gggcaaggtc gggagcggtt tcaccgatga agatttggtc 1440

gagttcacca agatgctcaa gcccctgatt gtccgtgaag agggcaagtt cgtcgagatt 1500

gagcccaagg tcgtcatcga ggtcacctac caggagatac agaagagccc caagtataag 1560

agcggtttcg cgcttcgctt cccgcgctac gtggcgctga gggaagataa aagcccggag 1620

gaggctgaca ccatagagag ggtcgcccag ctctacgagc tccaggagag gttcaaggcg 1680

aagaggtaat ctaga 1695

<210>2

<211>1695

<212>DNA

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>2

atgggcgata tgaggtactc cgagctggcc gaactctacc ggaggcttga gaagaccacg 60

ctcaaaacgc tcaagaccaa gttcgtcgcg gacttcctca agaaaacacc cgacgatttg 120

ctcgagatag ttccctacct gattctcggc aaggtctttc cggactggga cgagcgcgag 180

ctcggcgtcg gcgagaagct tctcataagg gccgtttcga tggcaaccgg cgtccccgag 240

agggaaatcg agaactcgat taaggacacc ggcgatttgg gcgagagcgt ggctctggct 300

ctaaagaaga ggaagcagaa gagcttcttc agccagcccc tcacgataaa gcgcgtctac 360

agcaccttcg ttaaggttgc cgaggcgagc ggagagggga gccaggacag gaagatgaag 420

tacttagcaa acctcttcat ggatgctcaa cccgaggagg gcaagtacat agccagaacc 480

gtcctcggga cgatgaggac gggcgtcgcc gagggaatcc tgcgcgatgc catagcggag 540

tccttcaagg tgaagccaga gctcgtcgag agggcctaca tgctcacgag cgacttcggc 600

tacgtggcga aggtcgccaa gctcgagggg aacgaggggc tctcgaaggt cagcatacag 660

attgggaagc cgataaggcc gatgctagct caaaacgccg ccaacgtcaa ggaagcgcta 720

atcgagatgg gcggtgaggc ggccttcgag attaagtacg acggcgcgcg cgttcaggtc 780

caccgcgatg gggacagggt gataatctac tcgaggaggc ttgagaacgt cacccgctcg 840

attcctgaga tagttgaggc ggtaaaggcc tccctgaagc cctctaatgt catagtcgag 900

ggcgagctgg ttgccgtcgg cgagaacggt cgcccgaggc ccttccagta cgtcctgagg 960

cgctttagga ggaagtacaa catcgaggag atgattgaga agattccgct cgagctcaac 1020

ctcttcgaca tcctctacgt tgaaggagag agcctcatcg acacgaagtt cgcagagagg 1080

aggaagaagc tcgaggagag cgttgaggag agcgataaga taaagctcgc cgaacagctc 1140

gttacgaaga aggttgaaga ggccgaggag ttctacaaga gggcccttga gctcggccac 1200

gagggcctga tggcgaagag gctggactcc atctacgagc ccggaaaccg cggtaagaag 1260

tggctgaaga ttaagcccac gatggagaac cttgacctcg tcattatcgg agccgaatgg 1320

ggcgagggca ggcgcgcgca cctgctcggc tcgttcctcg ttgcggccta cgaccccgag 1380

agcggtgagt tcgtcccggt gggcaaggtc gggagcggtt tcaccgatga agatttggtc 1440

gagttcacca agatgctcaa gcccctgatt gtccgtgaag agggcaagtt cgtcgagatt 1500

gagcccaagg tcgtcatcga ggtcacctac caggagatac agaagagccc caagtataag 1560

agcggtttcg cgcttcgctt cccgcgctac gtggcgctga gggaagataa aagcccggag 1620

gaggctgaca ccatagagag ggtcgcccag ctctacgagc tccaggagag gttcaaggcg 1680

aagaagtaat ctaga 1695

<210>3

<211>1695

<212>DNA

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>3

atgggcgata tgaggtactc cgagctggcc gaactttaca gaaggcttga gaagaccacg 60

ctcaaaacgc tcaagaccaa gtttgtcgcg gacttcctca agaaaactcc cgacgatttg 120

ctcgagatag ttccctacct gattctcggc aaggtctttc cggactggga cgagcgcgag 180

ctcggcgtcg gggaaaagtt gctcataagg gccgtttcga tggcaaccgg cgtccccgag 240

agggaaatcg agaactcgat taaggacacg ggcgatttgg gcgagagcgt ggctctggct 300

ctaaagaaga ggaagcagaa gagcttcttc agccagcccc tcacgataaa gcgcgtctac 360

agcaccttcg ttaaggttgc cgaggcgagc ggagagggga gccaggwcag gaagatgaag 420

tacttagcaa atctcttcat ggatgctcaa cccgaggagg gcaagtacat agccagaacc 480

gtcctcggaa cgatgaggac gggcgtcgcc gagggaatcc tgcgcgatgc catagcggag 540

gccttcaagg tgaagccaga gctcgtcgag agggcctaca tgctcacgag cgacttcggc 600

tacgtggcga aggtcgccaa gctcgagggg aacgaggggc tctcgaaggt cagcatacag 660

attgggaagc cgataaggcc gatgctagct caaaacgccg ccaacgtcaa ggaagcgcta 720

atcgagatgg gcggtgaggc ggccttcgag attaagtacg acggcgcgcg cgttcaggtc 780

caccgcgatg gggacagggt gataatctac tcgaggaggc ttgagaacgt cacccgctcg 840

attcctgaga tagttgaggc ggtaaaggcc tccctgaagc cctctaatgt catagtcgag 900

ggcgagctgg ttgccgtcgg cgagaacggt cgcccgaggc ccttccagta cgtcctgagg 960

cgctttagga ggaagtacaa catcgaggag atgattgaga agattccgct cgagctcaac 1020

ctcttcgaca tcctctacgt tgaaggggag agcctcattg acacgaagtt cgccgagagg 1080

aggaggaagc ttgaggagag cgtcgaggag ggcgataaga taaagctcgc cgaacagctc 1140

gttacgaaga aggtcgaaga ggccgaggag ttctacaaga gggcccttga gctcggccac 1200

gagggcctga tggcgaagag gctggactcc atctacgagc ccggaaaccg cggtaagaag 1260

tggctgaaga ttaagcccac gatggagaac cttgacctcg tcattatcgg agccgaatgg 1320

ggcgagggga ggcgcgcgca cctgcttggc tcgttcctcg ttgcggccta cgaccccgag 1380

agcggtgagt tcgtcccggt gggcaaggtc gggagcggtt tcaccgatga agatttggtc 1440

gagttcacca agatgctcaa gcccctgatt gtccgtgaag agggcaagtt cgtcgagatt 1500

gagcccaagg tcgtcatcga ggtcacctac caggagatac agaagagccc caagtataag 1560

agcggtttcg cgcttcgctt cccgcgctac gtggcgctga gggaagataa aagcccggag 1620

gaggctgaca ccatagagag ggtcgcccag ctctacgagc tccaggagag gttcaaggcg 1680

aagaagtaat ctaga 1695

<210>4

<211>1695

<212>DNA

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>4

atgggcgata tgaggtactc cgagctggcc gaactctacc ggaggcttga gaagaccacg 60

ctcaaaacgc tcaagaccaa gttcgtcgcg gacttcctca agaaaacacc cgacgatttg 120

ctcgagatag ttccctacct gattctcggc aaggtctttc cggactggga cgagcgcgag 180

ctcggcgtcg gggaaaagtt gctcataagg gccgtttcga tggcgactgg ggttccagag 240

agggaaatcg agaactcgat taaggacacg ggcgatttgg gcgagagcgt ggccttggcc 300

ctaaagaaga ggaagcagaa gagcttcttc agccagcccc tcacgataaa gcgcgtctac 360

agcaccttcg ttaaggttgc cgaggcgagc ggagagggga gccaggacag gaagatgaag 420

tacttagcaa acctcttcat ggatgctcaa cccgaggagg gcaagtacat agccagaacc 480

gtcctcggga cgatgaggac gggcgtcgcc gagggaatcc tgcgcgatgc catagcggag 540

gccttcaagg tgaagccaga gctcgtcgag agggcctaca tgctcacgag cgacttcggc 600

tacgtggcga aggttgcaaa gctcgagggg aacgaggggc tctcaaaggt tagcatacag 660

attgggaagc cgataaggcc gatgctagct caaaacgccg ccaacgtcaa ggaagcgcta 720

atcgagatgg gcggtgaggc ggccttcgag attaagtacg acggcgcgcg cgttcaggtc 780

caccgcgatg gggacagggt gataatctac tcgaggaggc ttgagaacgt cacccgctcg 840

attcctgaga tagttgaggc ggtaaaggcc tccctgaagc cctctaatgt catagtcgag 900

ggcgagctgg ttgccgtcgg cgagaacggt cgcccgaggc ccttccagta cgtcctgagg 960

cgctttagga ggaagtacaa catcgaggag atgattgaga agattccgct cgagctcaac 1020

ctcttcgaca tcctctacgt tgaaggagag agcctcatcg acacgaagtt cgcagagagg 1080

aggaagaagc tcgaggagag cgttgaggag agcgataaga taaagctcgc cgaacagctc 1140

gttacgaaga aggtcgaaga ggccgaggag ttctacaaga gggcccttga gctcggccac 1200

gagggcctga tggcgaagag gctggactcc atctacgagc ccggaaaccg cggtaagaag 1260

tggctgaaga ttaagcccac gatggagaac cttgacctcg tcattatcgg agccgaatgg 1320

ggcgagggga ggcgcgcgca cctgctcggc tcgttcctcg ttgcggccta cgaccccgag 1380

agcggtgagt tcgtcccggt gggcaaggtc gggagcggtt tcaccgatga agatttggtc 1440

gagttcacca agatgctcaa gcccctgatt gtccgtgaag agggcaagtt tgtcgagatt 1500

gagcccaagg tcgtcatcga ggtcacctac caggagatac agaagagccc caagtataag 1560

agcggtttcg cgcttcgctt cccgcgctac gtggcgctga gggaagataa aagcccggag 1620

gaggccgaca ccatagagag ggtcgcccag ctctacgagc tccaggagag gttcaaggcg 1680

aagaagtaat ctaga 1695

<210>5

<211>1695

<212>DNA

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>5

atgggcgata tgaggtactc cgagctggcc gaactctacc ggaggcttga gaagaccacg 60

ctcaaaacgc tcaagaccaa gttcgtcgcg gacttcctca agaaaacacc cgacgatttg 120

ctcgagatag ttccctacct gattctcggc aaggtctttc cggactggga cgagcgcgag 180

ctcggcgtcg gggaaaagtt gctcataagg gccgtttcga tggcgactgg ggttccagag 240

agggaaatcg agaactcgat taaggacacg ggcgatttgg gcgagagcgt ggccttggcc 300

ctaaagaaga ggaagcagaa gagcttcttc agccagcccc tcacgataaa gcgcgtctac 360

agcaccttcg ttaaggttgc cgaggcgagc ggagagggga gccaggacag gaagatgaag 420

tacttagcaa acctcttcat ggatgctcaa cccgaggagg gcaagtacat agccagaacc 480

gtcctcggga cgatgaggac gggcgtcgcc gagggaatcc tgcgcgatgc catagcggag 540

gccttcaagg tgaagccaga gctcgtcgag agggcctaca tgctcacgag cgacttcggc 600

tacgtggcga aggttgcaaa gctcgagggg aacgaggggc tctcaaaggt tagcatacag 660

attgggaagc cgataaggcc gatgctagct caaaacgccg ccaacgtcaa ggaagcgcta 720

atcgagatgg gcggtgaggc ggccttcgag attaagtacg acggcgcgcg cgttcaggtc 780

caccgcgacg gggatagggt gataatatac tcgaggaggc ttgagaacgt cacccgctcg 840

attcctgaga tagtcgaggc ggtaaaggcc tccctgaagc cttctaaggt catagtcgag 900

ggcgagctgg ttgccgtcgg cgagaacggt cgcccgaggc ccttccagta cgtcctcagg 960

aggttcagga ggaagtacaa catcgaggag atgattgaga agattccgct cgagctcaac 1020

ctcttcgata tcctctacgt tgatggggag agtctcatcg acacgaagtt cgcagagagg 1080

aggaagaaac tcgaggagag tgttgaggag agcgataaga taaagctcgc cgaacagctc 1140

gttacgaaga aggtcgaaga ggccgaggag ttctacaaga gggcccttga gctcggccac 1200

gagggcctga tggcgaagag gctggactcc atctacgagc ccggaaaccg cggtaagaag 1260

tggctgaaga ttaagcccac gatggagaac cttgacctcg tcattatcgg agccgaatgg 1320

ggcgagggga ggcgcgcgca cctgcttggc tcgttcctcg ttgcggccta cgaccccgag 1380

agcggtgagt tcgtcccggt gggcaaggtc gggagcggtt tcaccgatga agatttggtc 1440

gagttcacca agatgctcaa gcccctgatt gtccgtgaag agggcaagtt tgtcgagatt 1500

gagcccaagg tcgtcatcga agtaacgtat caggagatac agaagagccc caagtataag 1560

agcggtttcg cgcttcgctt cccgcgctac gtggcgctga gggaagataa aagcccggag 1620

gaggctgaca ccatagagag ggtcgcccag ctctacgagc tccaggagag gttcaaggcg 1680

aagaagtaat ctaga 1695

<210>6

<211>1695

<212>DNA

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>6

atgggcgaca tgaggtactc cgagctggcc gaactctacc ggaggcttga gaagaccacg 60

ctcaaaacgc tcaagaccaa gttcgtcgcg gacttcctca agaaaacacc cgacgatttg 120

ctcgagatag ttccctacct gattctcggc aaggtctttc cggactggga cgagcgcgag 180

ctcggcgtcg gggaaaagtt gctcataagg gccgtttcga tggcgactgg ggttccagag 240

agggaaatcg agaactcgat taaggacacg ggcgatttgg gcgagagcgt ggccttggcc 300

ctaaagaaga ggaagcagaa gagcttcttc agccagcccc tcacgataaa gcgcgtctac 360

agcaccttcg ttaaggttgc cgaggcgagc ggagagggga gccaggacag gaagatgaag 420

tacttagcaa acctcttcat ggatgctcaa cccgaggagg gcaagtacat agccagaacc 480

gtcctcggga cgatgaggac gggcgtcgcc gagggaatcc tgcgcgatgc catagcggag 540

gccttcaagg tgaagccaga gctcgtcgag agggcctaca tgctcacgag cgacttcggc 600

tacgtggcga aggttgcaaa gctcgagggg aacgaggggc tctcaaaggt tagcatacag 660

attgggaagc cgataaggcc gatgctagct caaaacgccg ccaacgtcaa ggaagcgcta 720

atcgagatgg gcggtgaggc ggccttcgag attaagtacg acggcgcacg cgttcaggtc 780

caccgcgacg gggacagggt gataatctac tcgaggaggc ttgagaacat cacccgctcg 840

attcctgaga tagtcgaggc ggtaaaggcc tccctgaagc cttctaaggt catagtcgag 900

ggcgagctgg ttgccgtcgg cgagaacggt cgcccgaggc ccttccagta cgtcctcagg 960

aggttcagga ggaagtacaa catcgaggag atgattgaga agattccgct cgagctcaac 1020

ctcttcgata tcctctacgt tgatggggag agtctcatcg acacgaagtt cgcagagagg 1080

aggaagaaac tcgaggagag tgttgaggag agcgataaga taaagctcgc cgaacagctc 1140

gttacgaaga aggtcgaaga ggccgaggag ttctacaaga gggcccttga gctcggccac 1200

gagggcctga tggcgaagag gctggactcc atctacgagc ccggaaaccg cggtaagaag 1260

tggctgaaga ttaagcccac gatggagaac cttgacctcg tcattatcgg agccgaatgg 1320

ggcgagggga ggcgcgcgca cctgcttggc tcgttcctcg ttgcggccta cgaccccgag 1380

agcggtgagt tcgtcccggt gggcaaggtc gggagcggtt tcaccgatga agatttggtc 1440

gagttcacca agatgctcaa gcccctgatt gtccgtgaag agggcaagtt tgtcgagatt 1500

gagcccaagg tcgtcatcga agtaacgtat caggagatac agaagagccc caagtataag 1560

agcggtttcg cgcttcgctt cccgcgctac gtggcgctga gggaagataa aagcccggag 1620

gaggctgaca ccatagagag ggtcgcccag ctctacgagc tccaggagag gtacaaggcg 1680

aagaggtaat ctaga 1695

<210>7

<211>1695

<212>DNA

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>7

atgggcgata tgaggtactc cgagctggcc gaactttaca gaaggcttga gaagaccacg 60

ctcaaaacgc tcaagaccaa gttcgtcgcg gacttcctca agaaaactcc cgacgatttg 120

ctcgagatag ttccctacct gattctcggc aaggtctttc cggactggga cgagcgcgag 180

ctcggcgtcg gggaaaagtt gctcataagg gccgtttcga tggcaaccgg cgtccccgag 240

agggaaatcg agaactcgat taaggacacg ggcgatttgg gcgagagcgt ggctctggct 300

ctaaagaaga ggaagcagaa gagcttcttc agccagcccc tcacgataaa gcgcgtctac 360

agcaccttcg ttaaggttgc cgaggcgagc ggagagggga gccaggacag gaagatgaag 420

tacttagcaa atctcttcat ggatgctcaa cccgaggagg gcaagtacat agccagaacc 480

gtcctcggaa cgatgaggac gggcgtcgcc gagggaatcc tgcgcgatgc catagcggag 540

gccttcaagg tgaagccaga gctcgtcgag agggcctaca tgctcacgag cgacttcgga 600

tacgtggcaa aggttgcaaa gctcgagggg aacgaggggc tctcaaaggt tagcatacag 660

attgggaagc cgataaggcc gatgctagct caaaacgccg ccaacgtcaa ggaagcgcta 720

atcgagatgg gcggtgaggc ggccttcgag attaagtacg acggtgcgcg cgttcaggtc 780

caccgcgacg gggatagggt gataatctac tcgaggaggc tcgagaacgt cacccgctcg 840

attcctgaga tagtcgaggc ggtaaaggcc tccctgaagc cttctaaggt catagtcgag 900

ggcgagctgg ttgccgtcgg cgagaacggt cgcccgaggc ccttccagta cgtcctcagg 960

aggttcagga ggaagtacaa catcgaggag atgattgaga agattccgct cgagctcaac 1020

ctcttcgata tcctctacgt tgatggggag agtctcatcg acacgaagtt cgcagagagg 1080

aggaagaaac tcgaggagag tgttgaggag agcgataaga taaagctcgc cgaacagctc 1140

gttacgaaga aggtcgaaga ggccgaggag ttctacaaga gggcccttga gctcggccac 1200

gagggcctga tggcgaagag gctggactcc atctacgagc ccggaaaccg cggtaagaag 1260

tggctgaaga ttaagcccac gatggagaac cttgacctcg tcattatcgg agccgaatgg 1320

ggcgagggga ggcgcgcgca cctgcttggc tcgttcctcg ttgcggccta cgaccccgag 1380

agcggtgagt tcgtcccggt gggcaaggtc gggagcggtt tcaccgatga agatttggtc 1440

gagttcacca agatgctcaa gcccctgatt gtccgtgaag agggcaagtt cgtcgagatt 1500

gagcccaagg tcgtcatcga ggtcacctac caggagatac agaagagccc caagtataag 1560

agcggtttcg cgcttcgctt cccgcgctac gtggcgctga gggaagataa aagcccggag 1620

gaggctgaca ccatagagag ggtcgcccag ctctacgagc tccaggagag gttcaaggcg 1680

aagaagtaat ctaga 1695

<210>8

<211>564

<212>PRT

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>8

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ala Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Val Ser Ile Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Arg Asp Gly Asp Arg Val Ile Ile Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val

275 280 285

Lys Ala Ser Leu Lys Pro Ser Lys Val Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Asp Gly Glu Ser Leu

340 345 350

Ile Asp Thr Lys Phe Ala Glu Arg Arg Lys Lys Leu Glu Glu Ser Val

355 360 365

Glu Glu Ser Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe

450 455 460

Val Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

Ile Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala

545 550 555 560

Lys Lys Ser Arg

<210>9

<211>564

<212>PRT

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>9

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ala Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Va lSer Ile Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Arg Asp Gly Asp Arg ValIle Ile Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val

275 280 285

Lys Ala Ser Leu Lys Pro Ser Lys Val Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Asp Gly Glu Ser Leu

340 345 350

Ile Asp Thr Lys Phe Ala Glu Arg Arg Lys Lys Leu Glu Glu Ser Val

355 360 365

Glu Glu Ser Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe

450 455 460

Val Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

Ile Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala

545 550 555 560

Lys Lys Ser Arg

<210>10

<211>564

<212>PRT

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>10

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ala Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Val Ser Ile Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Arg Asp Gly Asp Arg Val Ile Ile Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val

275 280 285

Lys Ala Ser Leu Lys Pro Ser Asn Val Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Glu Gly Glu Ser Leu

340 345 350

Ile Asp Thr Lys Phe Ala Glu Arg Arg Lys Lys Leu Glu Glu Ser Val

355 360 365

Glu Glu Ser Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe

450 455 460

Val Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

Ile Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala

545 550 555 560

Lys Lys Ser Arg

<210>11

<211>563

<212>PRT

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>11

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Arg Lys Met Lys Tyr Leu Ala Asn Leu

130 135 140

Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr Val

145 150 155 160

Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp Ala

165 170 175

Ile Ala Glu Ala Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala Tyr

180 185 190

Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu Glu

195 200 205

Gly Asn Glu Gly Leu Ser Lys Val Ser Ile Gln Ile Gly Lys Pro Ile

210 215 220

Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu Ile

225 230 235 240

Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala Arg

245 250 255

Val Gln Val His Arg Asp Gly Asp Arg Val Ile Ile Tyr Ser Arg Arg

260 265 270

Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val Lys

275 280 285

Ala Ser Leu Lys Pro Ser Asn Val Ile Val Glu Gly Glu Leu Val Ala

290 295 300

Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg Arg

305 310 315 320

Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro Leu

325 330 335

Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Glu Gly Glu Ser Leu Ile

340 345 350

Asp Thr Lys Phe Ala Glu Arg Arg Arg Lys Leu Glu Glu Ser Val Glu

355 360 365

Glu Gly Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys Val

370 375 380

Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His Glu

385 390 395 400

Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn Arg

405 410 415

Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp Leu

420 425 430

Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu Leu

435 440 445

Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe Val

450 455 460

Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val Glu

465 470 475 480

Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys Phe

485 490 495

Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu Ile

500 505 510

Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro Arg

515 520 525

Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr Ile

530 535 540

Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala Lys

545 550 555 560

Lys Ser Arg

<210>12

<211>564

<212>PRT

<213>未知

<220>

<223>Thermoccocus sp.strain 9 degrees North

<400>12

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ser Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Val Ser Ile Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Arg Asp Gly Asp Arg Val Ile Ile Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val

275 280 285

Lys Ala Ser Leu Lys Pro Ser Asn Val Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Glu Gly Glu Ser Leu

340 345 350

Ile Asp Thr Lys Phe Ala Glu Arg Arg Lys Lys Leu Glu Glu Ser Val

355 360 365

Glu Glu Ser Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe

450 455 460

Val Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

Ile Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala

545 550 555 560

Lys Arg Ser Arg

<210>13

<211>564

<212>PRT

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>13

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ser Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Val Ser Ile Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Arg Asp Gly Asp Arg Val Ile Ile Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val

275 280 285

Lys Ala Ser Leu Lys Pro Ser Asn Val Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Glu Gly Glu Ser Leu

340 345 350

Ile Asp Thr Lys Phe Ala Glu Arg Arg Lys Lys Leu Glu Glu Ser Val

355 360 365

Glu Glu Ser Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe

450 455 460

Val Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Al a Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

lle Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala

545 550 555 560

Lys Lys Ser Arg

<210>14

<211>564

<212>PRT

<213>未知

<220>

<223>嗜热球菌属某种菌株9°N

<400>14

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ala Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Val Ser Ile Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Arg Asp Gly Asp Arg Val Ile Ile Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Ile Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val

275 280 285

Lys Ala Ser Leu Lys Pro Ser Lys Val Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Asp Gly Glu Ser Leu

340 345 350

Ile Asp Thr Lys Phe Ala Glu Arg Arg Lys Lys Leu Glu Glu Ser Val

355 360 365

Glu Glu Ser Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe

450 455 460

Val Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

Ile Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Tyr Lys Ala

545 550 555 560

Lys Arg Ser Arg

<210>15

<211>564

<212>PRT

<213>未知

<220>

<223>嗜热球菌属某种菌株9°NB2

<400>15

Met Gly Asp Met Arg Tyr Ser Glu Leu Ala Glu Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Asp Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Arg Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Arg Glu Ile Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Leu Lys Lys Arg Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Ser Thr Phe Val Lys Val Ala Glu

115 120 125

Ala Ser Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Gln Pro Glu Glu Gly Lys Tyr Ile Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ser Phe Lys Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Val Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Val Ser Ile Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Asn Val Lys Glu Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Arg Asp Gly Asp Arg Val Ile Ile Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val

275 280 285

Lys Ala Ser Leu Lys Pro Ser Asn Val Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Ile Leu Tyr Val Glu Gly Glu Ser Leu

340 345 350

Ile Asp Thr Lys Phe Ala Glu Arg Arg Lys Lys Leu Glu Glu Ser Val

355 360 365

Glu Glu Ser Asp Lys Ile Lys Leu Ala Glu Gln Leu Val Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Glu Phe Tyr Lys Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro Glu Ser Gly Glu Phe

450 455 460

Val Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

Ile Glu Arg Val Ala Gln Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala

545 550 555 560

Lys Lys Ser Arg

<210>16

<211>559

<212>PRT

<213>未知

<220>

<223>喜烟嗜热球菌(Thermococcus sp.tfumicolans)

<400>16

Met Lys Tyr Ser Glu Leu Ala Gly Leu Tyr Arg Arg Leu Glu Lys Thr

1 5 10 15

Thr Leu Lys Thr Leu Lys Thr Arg Phe Val Ala Asp Phe Leu Lys Asn

20 25 30

Val Pro Asp Glu Leu Leu Glu Ile Val Pro Tyr Leu Ile Leu Gly Lys

35 40 45

Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly Glu Lys Leu

50 55 60

Leu Ile Lys Ala Val Ser Ile Ala Thr Gly Val Pro Glu Gly Glu Ile

65 70 75 80

Glu Asn Ser Ile Lys Asp Thr Gly Asp Leu Gly Glu Ser Ile Ala Leu

85 90 95

Ala Val Lys Lys Lys Lys Gln Lys Ser Phe Phe Ser Gln Pro Leu Thr

100 105 110

Ile Lys Arg Val Tyr Asp Thr Phe ValLys Val Ala Glu Ser Gln Gly

115 120 125

Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn Leu Phe Met

130 135 140

Asp Ala Gln Pro Glu Glu Ala Lys Tyr Ile Ala Arg Thr Val Leu Gly

145 150 155 160

Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp Ala Ile Ala

165 170 175

Glu Ala Phe Lys Val Lys Ala Glu Leu Val Glu Arg Ala Tyr Met Leu

180 185 190

Thr Ser Asp Phe Gly Tyr Val Thr Lys Val Ala Lys Leu Glu Gly Asn

195 200 205

Glu Gly Leu Ser Lys Val Arg Ile Gln Val Gly Lys Pro Val Arg Pro

210 215 220

Met Leu Ala Gln Asn Ala Ala Ser Val Lys Asp Ala Leu Leu Glu Met

225 230 235 240

Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala Arg Val Gln

245 250 255

Val His Lys Asp Gly Asp Arg Val Val Ile Tyr Ser Arg Arg Leu Glu

260 265 270

Asn Val Thr Arg Ser Ile Pro Glu Ile Val Glu Ala Val Arg Ser Gln

275 280 285

Leu Arg Pro Glu Lys Ala Ile Val Glu Gly Glu Leu ValAla Val Gly

290 295 300

Asp Gly Gly Lys Pro Arg Pro Phe Gln Tyr Val Leu Arg Arg Phe Arg

305 310 315 320

Arg Lys Tyr Asn Ile Glu Glu Met Ile Glu Arg Ile Pro Leu Glu Leu

325 330 335

Asn Leu Phe Asp Val Leu Tyr Val Asp Gly Glu Ser Leu Val Asp Thr

340 345 350

Pro Phe Met Glu Arg Arg Lys Arg Leu Glu Glu Ala Val Glu Glu Ser

355 360 365

Glu Arg Ile Lys Leu Ala Gln Gln Leu Val Thr Lys Lys Ala Glu Glu

370 375 380

Ala Glu Glu Phe Tyr Arg Arg Ala Leu Glu Leu Gly His Glu Gly Leu

385 390 395 400

Met Ala Lys Arg Leu Asp Ser Val Tyr Glu Pro Gly Asn Arg Gly Lys

405 410 415

Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asp Leu Asp Leu Val Ile

420 425 430

Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala Hi s Leu Leu Gly Ser

435 440 445

Phe Leu Val Ala Ala Tyr Asp Gln His Arg Gly Glu Phe Val Pro Val

450 455 460

Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Ala Glu Phe Thr

465 470 475 480

Lys Met Leu Lys Pro Leu Ile Val Arg Glu Glu Gly Lys Tyr Val Glu

485 490 495

Ile Glu Pro Arg Val Val Ile Gln Val Thr Tyr Gln Glu Ile Gln Lys

500 505 510

Ser Pro Lys Tyr Glu Ser Gly Phe Ala Leu Arg Phe Pro Arg Tyr Val

515 520 525

Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr Ile Glu Arg

530 535 540

Ile Ser Glu Leu Tyr Gly Leu Gln Glu Arg Phe Lys Ala Lys Arg

545 550 555

<210>17

<211>562

<212>PRT

<213>未知

<220>

<223>克达卡拉嗜热球菌(Thermococcus sp.tkodakaraenis)

<400>17

Met Ser Asp Met Arg Tyr Ser Glu Leu Ala Asp Leu Tyr Arg Arg Leu

1 5 10 15

Glu Lys Thr Thr Leu Lys Thr Leu Lys Thr Lys Phe Val Ala Asp Phe

20 25 30

Leu Lys Lys Thr Pro Asp Glu Leu Leu Glu Ile Val Pro Tyr Leu Ile

35 40 45

Leu Gly Lys Val Phe Pro Asp Trp Asp Glu Arg Glu Leu Gly Val Gly

50 55 60

Glu Lys Leu Leu Ile Lys Ala Val Ser Met Ala Thr Gly Val Pro Glu

65 70 75 80

Lys Glu Ile Glu Asp Ser Val Arg Asp Thr Gly Asp Leu Gly Glu Ser

85 90 95

Val Ala Leu Ala Ile Lys Lys Lys Lys Gln Lys Ser Phe Phe Ser Gln

100 105 110

Pro Leu Thr Ile Lys Arg Val Tyr Asp Thr Phe Val Lys Ile Ala Glu

115 120 125

Ala Gln Gly Glu Gly Ser Gln Asp Arg Lys Met Lys Tyr Leu Ala Asn

130 135 140

Leu Phe Met Asp Ala Glu Pro Glu Glu Gly Lys Tyr Leu Ala Arg Thr

145 150 155 160

Val Leu Gly Thr Met Arg Thr Gly Val Ala Glu Gly Ile Leu Arg Asp

165 170 175

Ala Ile Ala Glu Ala Phe Arg Val Lys Pro Glu Leu Val Glu Arg Ala

180 185 190

Tyr Met Leu Thr Ser Asp Phe Gly Tyr Val Ala Lys Ile Ala Lys Leu

195 200 205

Glu Gly Asn Glu Gly Leu Ser Lys Val ArgIle Gln Ile Gly Lys Pro

210 215 220

Ile Arg Pro Met Leu Ala Gln Asn Ala Ala Ser Val Lys Asp Ala Leu

225 230 235 240

Ile Glu Met Gly Gly Glu Ala Ala Phe Glu Ile Lys Tyr Asp Gly Ala

245 250 255

Arg Val Gln Val His Lys Asp Gly Asp Lys Val Ile Val Tyr Ser Arg

260 265 270

Arg Leu Glu Asn Val Thr Arg Ser Ile Pro Glu Val Ile Glu Ala Ile

275 280 285

Lys Ala Ala Leu Lys Pro Glu Lys Ala Ile Val Glu Gly Glu Leu Val

290 295 300

Ala Val Gly Glu Asn Gly Arg Pro Arg Pro Phe Gln Tyr Val Leu Arg

305 310 315 320

Arg Phe Arg Arg Lys Tyr Asn Ile Asp Glu Met Ile Glu Lys Ile Pro

325 330 335

Leu Glu Leu Asn Leu Phe Asp Val Met Phe Val Asp Gly Glu Ser Leu

340 345 350

Ile Glu Thr Lys Phe Ile Asp Arg Arg Asn Lys Leu Glu Glu Ile Val

355 360 365

Lys Glu Ser Glu Lys Ile Lys Leu Ala Glu Gln Leu Ile Thr Lys Lys

370 375 380

Val Glu Glu Ala Glu Ala Phe TyrArg Arg Ala Leu Glu Leu Gly His

385 390 395 400

Glu Gly Leu Met Ala Lys Arg Leu Asp Ser Ile Tyr Glu Pro Gly Asn

405 410 415

Arg Gly Lys Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp

420 425 430

Leu Val Ile Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu

435 440 445

Leu Gly Ser Phe Leu Val Ala Ala Tyr Asp Pro His Ser Gly Glu Phe

450 455 460

Leu Pro Val Gly Lys Val Gly Ser Gly Phe Thr Asp Glu Asp Leu Val

465 470 475 480

Glu Phe Thr Lys Met Leu Lys Pro Tyr Ile Val Arg Gln Glu Gly Lys

485 490 495

Phe Val Glu Ile Glu Pro Lys Phe Val Ile Glu Val Thr Tyr Gln Glu

500 505 510

Ile Gln Lys Ser Pro Lys Tyr Lys Ser Gly Phe Ala Leu Arg Phe Pro

515 520 525

Arg Tyr Val Ala Leu Arg Glu Asp Lys Ser Pro Glu Glu Ala Asp Thr

530 535 540

Ile Glu Arg Val Ala Glu Leu Tyr Glu Leu Gln Glu Arg Phe Lys Ala

545 550 555 560

Lys Lys

<210>18

<211>559

<212>PRT

<213>未知

<220>

<223>深海热球菌(Pyrococcus sp.pabyssi)

<400>18

Met Arg Tyr Ile Glu Leu Ala Gln Leu Tyr Gln Lys Leu Glu Lys Thr

1 5 10 15

Thr Met Lys Leu Ile Lys Thr Arg Leu Val Ala Asp Phe Leu Lys Lys

20 25 30

Val Pro Glu Asp His Leu Glu Phe Ile Pro Tyr Leu Ile Leu Gly Asp

35 40 45

Val Phe Pro Glu Trp Asp Glu Arg Glu Leu Gly Val Gly Glu Lys Leu

50 55 60

Leu Ile Lys Ala Val Ser Met Ala Thr Gly Ile Asp Ser Lys Glu Ile

65 70 75 80

Glu Asn Ser Val Lys Asp Thr Gly Asp Leu Gly Glu Ser Ile Ala Leu

85 90 95

Ala Val Lys Arg Arg Lys Gln Lys Ser Phe Phe Ser Gln Pro Leu Thr

100 105 110

Ile Lys Arg Val Tyr Gln Thr Leu Val Lys Val Ala Glu Thr Thr Gly

115 120 125

Glu Gly Ser Gln Asp Lys Lys Met Lys Tyr Leu Ala Asn Leu Phe Met

130 135 140

Asp Ala Glu Pro Ile Glu Ala Lys Tyr Ile Ala Arg Thr Val Leu Gly

145 150 155 160

Thr Met Arg Thr Gly Val Ala Glu Gly Leu Leu Arg Asp Ala Ile Ser

165 170 175

Leu Ala Phe Asn Val Lys Val Glu Leu Val Glu Arg Ala Tyr Met Leu

180 185 190

Thr Ser Asp Phe Gly Phe Val Ala Lys Ile Ala Lys Thr Glu Gly Asn

195 200 205

Asp Gly Leu Ala Lys Val Thr Ile Gln Ile Gly Lys Pro Ile Lys Pro

210 215 220

Met Leu Ala Gln Gln Ala Ala Asn Ile Lys Glu Ala Leu Leu Glu Met

225 230 235 240

Gly Gly Glu Ala Glu Phe Glu Ile Lys Tyr Asp Gly Ala Arg Val Gln

245 250 255

Val His Lys Asp Gly Glu Lys Val Thr Ile Tyr Ser Arg Arg Leu Glu

260 265 270

Asn Val Thr Arg Ala Ile Pro Glu Ile Val Glu Ala Ile Lys Glu Ala

275 280 285

Leu Lys Pro Ala Lys Ala Ile Val Glu Gly Glu Leu Val Ala Ile Gly

290 295 300

Glu Asp Gly Arg Pro Leu Pro Phe Gln Tyr Val Leu Arg Arg Phe Arg

305 310 315 320

Arg Lys Tyr Asn Ile Glu Glu Met Met Glu Lys Ile Pro Leu Glu Leu

325 330 335

Asn Leu Phe Asp Val Leu Tyr Val Asp Gly Val Ser Leu Ile Asp Thr

340 345 350

Lys Phe Met Glu Arg Arg Lys Lys Leu Glu Glu Ile Val Glu Ala Asn

355 360 365

Gly Lys Val Lys Ile Ala Glu Asn Leu Ile Thr Lys Asn Val Glu Glu

370 375 380

Ala Glu Gln Phe Tyr Lys Arg Ala Leu Glu Met Gly His Glu Gly Leu

385 390 395 400

Met Ala Lys Arg Leu Asp Ala Ile Tyr Glu Pro Gly Asn Arg Gly Lys

405 410 415

Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp Leu Val Ile

420 425 430

Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu Leu Gly Ser

435 440 445

Phe Ile Leu Gly Ala Tyr Asp Pro Glu Thr Gly Glu Phe Leu Glu Val

450 455 460

Gly Lys Val Gly Ser Gly Phe Thr Asp Asp Asp Leu Val Glu Phe Thr

465 470 475 480

Lys Met Leu Arg Pro Leu Ile Ile Lys Glu Glu Gly Lys Arg Val Trp

485 490 495

Ile Gln Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu Ile Gln Lys

500 505 510

Ser Pro Lys Tyr Arg Ser Gly Phe Ala Leu Arg Phe Pro Arg Tyr Val

515 520 525

Ala Leu Arg Glu Asp Lys Gly Pro Glu Asp Ala Asp Thr Ile Glu Arg

530 535 540

Ile Ala Gln Leu Tyr Glu Leu Gln Glu Arg Met Lys Gly Lys Val

545 550 555

<210>19

<211>561

<212>PRT

<213>未知

<220>

<223>激烈热球菌(Pyrococcus sp.pfuriosus)

<400>19

Met Arg Tyr Leu Glu Leu Ala Gln Leu Tyr Gln Lys Leu Glu Lys Thr

1 5 10 15

Thr Met Lys Leu Ile Lys Thr Arg Leu Val Ala Asp Phe Leu Lys Lys

20 25 30

Val Pro Asp Asp His Leu Glu Phe Ile Pro Tyr Leu Ile Leu Gly Glu

35 40 45

Val Phe Pro Glu Trp Asp Glu Arg Glu Leu Gly Val Gly Glu Lys Leu

50 55 60

Leu Ile Lys Ala Val Ala Met Ala Thr Gly Ile Asp Ala Lys Glu Ile

65 70 75 80

Glu Glu Ser Val Lys Asp Thr Gly Asp Leu Gly Glu Ser Ile Ala Leu

85 90 95

Ala Val Lys Lys Lys Lys Gln Lys Ser Phe Phe Ser Gln Pro Leu Thr

100 105 110

Ile Lys Arg Val Tyr Gln Thr Leu Val Lys Val Ala Glu Thr Thr Gly

115 120 125

Glu Gly Ser Gln Asp Lys Lys Val Lys Tyr Leu Ala Asp Leu Phe Met

130 135 140

Asp Ala Glu Pro Leu Glu Ala Lys Tyr Leu Ala Arg Thr Ile Leu Gly

145 150 155 160

Thr Met Arg Thr Gly Val Ala Glu Gly Leu Leu Arg Asp Ala Ile Ala

165 170 175

Met Ala Phe His Val Lys Val Glu Leu Val Glu Arg Ala Tyr Met Leu

180 185 190

Thr Ser Asp Phe Gly Tyr Val Ala Lys Ile Ala Lys Leu Glu Gly Asn

195 200 205

Glu Gly Leu Ala Lys Val Gln Val Gln Leu Gly Lys Pro Ile Lys Pro

210 215 220

Met Leu Ala Gln Gln Ala Ala Ser Ile Arg Asp Ala Leu Leu Glu Met

225 230 235 240

Gly Gly Glu Ala Glu Phe Glu Ile Lys Tyr Asp Gly Ala Arg Val Gln

245 250 255

Val His Lys Asp Gly Ser Lys Ile Ile Val Tyr Ser Arg Arg Leu Glu

260 265 270

Asn Val Thr Arg Ala Ile Pro Glu Ile Val Glu Ala Leu Lys Glu Ala

275 280 285

Ile Ile Pro Glu Lys Ala Ile Val Glu Gly Glu Leu Val Ala Ile Gly

290 295 300

Glu Asn Gly Arg Pro Leu Pro Phe Gln Tyr Val Leu Arg Arg Phe Arg

305 310 315 320

Arg Lys His Asn Ile Glu Glu Met Met Glu Lys Ile Pro Leu Glu Leu

325 330 335

Asn Leu Phe Asp Val Leu Tyr Val Asp Gly Gln Ser Leu Ile Asp Thr

340 345 350

Lys Phe Ile Asp Arg Arg Arg Thr Leu Glu Glu Ile Ile Lys Gln Asn

355 360 365

Glu Lys Ile Lys Val Ala Glu Asn Leu Ile Thr Lys Lys Val Glu Glu

370 375 380

Ala Glu Ala Phe Tyr Lys Arg Ala Leu Glu Met Gly His Glu Gly Leu

385 390 395 400

Met Ala Lys Arg Leu Asp Ala Val Tyr Glu Pro Gly Asn Arg Gly Lys

405 410 415

Lys Trp Leu Lys Ile Lys Pro Thr Met Glu Asn Leu Asp Leu Val Ile

420 425 430

Ile Gly Ala Glu Trp Gly Glu Gly Arg Arg Ala His Leu Phe Gly Ser

435 440 445

Phe Ile Leu Gly Ala Tyr Asp Pro Glu Thr Gly Glu Phe Leu Glu Val

450 455 460

Gly Lys Val Gly Ser Gly Phe Thr Asp Asp Asp Leu Val Glu Phe Thr

465 470 475 480

Lys Met Leu Lys Pro Leu Ile Ile Lys Glu Glu Gly Lys Arg Val Trp

485 490 495

Leu Gln Pro Lys Val Val Ile Glu Val Thr Tyr Gln Glu Ile Gln Lys

500 505 510

Ser Pro Lys Tyr Arg Ser Gly Phe Ala Leu Arg Phe Pro Arg Phe Val

515 520 525

Ala Leu Arg Asp Asp Lys Gly Pro Glu Asp Ala Asp Thr Ile Glu Arg

530 535 540

Ile Ala Gln Leu Tyr Glu Leu Gln Glu Lys Met Lys Gly Lys Val Glu

545 550 555 560

Ser

<210>20

<211>31

<212>DNA

<213>未知

<220>

<223>引物

<220>

<221>misc_feature

<222>(15)..(15)

<223>r＝g or a

<220>

<221>misc_feature

<222>(20)..(20)

<223>y＝t/u or c

<220>

<221>misc_feature

<222>(24)..(24)

<223>m＝a or c

<220>

<221>misc_feature

<222>(25)..(25)

<223>r＝g or a

<220>

<221>misc_feature

<222>(26)..(26)

<223>s＝g or c

<400>20

cggtggtgca tatgrgcgay atgmrstact c 31

<210>21

<211>40

<212>DNA

<213>未知

<220>

<223>引物

<220>

<221>misc_feature

<222>(17)..(17)

<223>y＝t/u or c

<400>21

ataaactcta gattacytct tcgccttgaa cctctcctgg 40

<210>22

<211>39

<212>DNA

<213>未知

<220>

<223>引物

<220>

<221>misc_feature

<222>(20)..(20)

<223>y＝t/u or c

<400>22

cggtggtgca tatgggcgay atgaggtact ccgagctgg 39

<210>23

<211>23

<212>DNA

<213>未知

<220>

<223>引物

<400>23

cgaacgtcgc gcagagaaac agg 23

<210>24

<211>20

<212>DNA

<213>未知

<220>

<223>引物

<400>24

cctgctctgc cgcttcacgc 20

Claims

1.基本上纯的重组蛋白质，其具有DNA连接酶活性，并且与SEQID NO：13具有至少91％的氨基酸序列同一性。

2.基本上纯的蛋白质，其具有DNA连接酶活性，由选自下列的DNA序列编码：

(a)与SEQ ID NO：2基本上相同的序列；

(b)与SEQ ID NO：2基本上互补的序列；

(c)在严格条件下，与SEQ ID NO：2杂交的序列；和

(d)编码SEQ ID NO：13的序列。

3.根据权利要求1所述的蛋白质，其中在大约100℃的温度下温育30分钟后，保留了至少25％的连接酶活性。

4.根据权利要求2所述的蛋白质，其中在大约100℃的温度下温育30分钟后，保留了至少25％的连接酶活性。

5.根据权利要求1、2、3或4所述的蛋白质，其在连接过程中利用ATP而不是NAD⁺作为辅因子。

6.编码DNA连接酶的DNA，所述DNA具有选自下列的序列：

(a)与SEQ ID NO：2基本上相同的序列；

(b)与SEQ ID NO：2基本上互补的序列；

(c)在严格条件下，与SEQ ID NO：2杂交的序列；和

(d)编码SEQ ID NO：13的序列。

7.载体，含有权利要求6所述DNA。

8.宿主细胞，其能表达权利要求1所述的蛋白质。

9.连接磷酸二酯键的方法，包括：

(a)选择根据权利要求1或2所述的连接酶；

(b)将所述连接酶与DNA混合，所述DNA在所述DNA的至少一条链上含有切点；和

(c)在所述切点连接所述磷酸二酯键。

10.根据权利要求9所述的方法，其中所述连接酶是从古细菌分离物得到的热稳定连接酶。

11.根据权利要求9所述的方法，其中所述古细菌分离物为嗜热球菌属某种(Thermococcus sp.)(菌株9°N-7)。