CN114269767A - 利用嗜热蛋白体外转录和翻译重组的系统、方法及合成物 - Google Patents

利用嗜热蛋白体外转录和翻译重组的系统、方法及合成物 Download PDF

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CN114269767A
CN114269767A CN202080042247.3A CN202080042247A CN114269767A CN 114269767 A CN114269767 A CN 114269767A CN 202080042247 A CN202080042247 A CN 202080042247A CN 114269767 A CN114269767 A CN 114269767A
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迈克尔·亨伯特
亚历山大·科格林
查理·维拉纽瓦
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Abstract

本发明的另一个目的可包括一种重组无细胞表达系统，其中，反应混合物包含生物化合物、蛋白、酶、生物仿制药体外生物合成或小分子化学修饰所需的所有无细胞反应组分。

Description

利用嗜热蛋白体外转录和翻译重组的系统、方法及合成物
本申请要求2019年4月12日提交的第62/833,555号美国临时申请的优先权和权益。上述申请的整个说明书和附图通过本发明的整体引用，成为本发明的一部分。
技术领域
本发明涉及重组无细胞表达系统以及利用此系统实现生物材料体外高产的方法。
序列列表
本申请包含一个序列列表，此序列列表采用ASCII格式通过电子方式提交，并通过本发明的整体引用成为本发明的一部分。所述ASCII副本文件创建于2020年4月13日，文件名为90125.00096-Sequence-Listing_ST25.txt，文件大小为427千字节。
背景技术
无细胞表达系统(也称为体外转录/翻译系统、无细胞蛋白表达系统、无细胞翻译系统或无细胞生物合成系统)是一种使研究人员能够体外表达功能性蛋白或其他靶分子的分子生物学技术。此类系统可体外表达很难在体内产生的蛋白或其他小分子，还可高通量生产蛋白库，供蛋白进化、功能基因组学和结构研究使用。此类系统的另一个优势在于，需表达的靶蛋白通常可能对宿主细胞有毒，或通常与细胞表达不相容，使体内系统成为不适用但并非完全无效的蛋白表达载体。与基于细菌或组织培养细胞的体内技术相比，体外蛋白表达更快，因为这种表达无需进行基因转染、细胞培养或广泛蛋白纯化。
更具体地说，无细胞表达系统无需使用活细胞，即可产生靶分子和复合物(如RNA种属和蛋白)。典型的无细胞表达系统可利用细胞裂解物中发现的生物组分/机制，从包含一个或多个靶基因的DNA中产生靶分子。典型无细胞表达系统的常见组分可包括通常由细胞培养裂解物衍生的细胞提取物、一个能量源(如ATP)、一定量的氨基酸、辅因子(如镁)以及包含所需基因的核酸合成模板，此类模板通常采用质粒合成模板或线性表达(或合成)模板(LET或LST)的形式。可通过离心法或其他沉淀法裂解目标细胞并去除细胞壁、基因组DNA及其他碎片，获得细胞提取物。裂解物或细胞提取物的剩余部分可包含靶分子表达所需的必要细胞机制。
常见的无细胞表达系统涉及无细胞蛋白合成(CFPS)技术。为了生产一种或多种目标蛋白，典型的CFPS系统采用一系列必要的催化组分从微生物、植物或动物细胞的粗裂解物中产生能量并合成蛋白。粗裂解物包含DNA到RNA的转录、RNA到蛋白的翻译、蛋白折叠和能量代谢所需的必要元素(例如，核糖体、氨酰-tRNA合成酶、翻译起始因子和延伸因子、核糖体释放因子、核苷酸循环酶、代谢酶、分子伴侣和折叠酶等)。如今使用的常见细胞提取物由大肠杆菌(ECE)、兔网织红细胞(RRL)、麦芽(WGE)和昆虫细胞(ICE)，甚至哺乳动物细胞(MC)制成。
与常规体内蛋白表达方法相比，无细胞表达系统提供了诸多优势。无细胞系统可直接将细胞的大多数(即便不是全部)代谢资源专门用于生产一种蛋白。此外，体外缺少细胞壁和膜组分很有利，因为这样便可以控制合成环境。例如，为了反映所表达基因的密码子应用情况，可改变tRNA水平。与体内方法相比，由于不必太多关注细胞生长和活性情况，还可以更灵活地改变氧化还原电位、pH或离子强度。此外，很容易直接回收经纯化、正确折叠的蛋白产物。
尽管无细胞表达系统具有诸多有利方面，但是，之前的一些障碍限制其作为蛋白生产技术的应用。在基于大肠杆菌提取物的无细胞系统(在第7,118,883号美国专利中确定)和基于酵母提取物的无细胞系统(在第9,528,137号美国专利中确定)中特别提到这些障碍，其包括活性蛋白合成的反应持续时间短、蛋白生产率低、反应规模小、正确折叠包含多个二硫键的蛋白的能力有限，以及最初将其作为“黑箱”技术开发。因此，需要一种经济上可行的商用无细胞表达系统来增加产物产量、增强组分稳定性、提高蛋白生产率并延长反应时间。
如上所述，由于缺乏一致性、产量和规模化的可能性，无细胞系统并未广泛用于生产生物制剂。发明人之前报告了一种基于提取物的无细胞系统，这种系统利用示例性嗜热菌，通过热稳定蛋白代替大肠杆菌机制，改善了此类系统的应用，从而提高了生产率和产量，而且还包括一种新型能量再生系统。(第PCT/US201 8/012121号PCT申请对此类新型能量再生系统进行了一般说明，其说明、附图、示例、序列和权利要求通过本发明的整体引用，成为本发明的一部分。)
如下所述，发明人开发了一种完全重组体外转录/翻译系统，在一些实施例中，所述系统包含各种示例性嗜热菌中的肽基组分。如上所述，本商用无细胞系统基于添加来自大肠杆菌细胞提取物或基于重组大肠杆菌酶的必要转录/翻译机制。此外，还报告了各种其他提取物来源，包括利用嗜热菌改善体外蛋白生产，但目前尚未报告一种完全重组表达系统，包括基于嗜热蛋白的完全重组表达系统。
本创新性技术克服了传统无细胞表达系统的局限性，同时实现了一种真正极度高效、稳健的体外无细胞表达系统，可延长反应持续时间并提高产物产量(详细讨论如下)。具体地说，本发明包括一种基于嗜热菌的无细胞系统，此系统可重组表达转录/翻译所需的每种蛋白，从而实现连续流动，更好的控制并微调系统，并且不会遇到在基于提取物的成批系统中观察到的大量变量。此系统可用于初步研究应用中的小规模蛋白生产操作以及小型动物研究等中等规模应用。本发明允许在本文所述的新型生物反应器中采用连续流动方法实现大规模生产，并代替占地面积和人员需求更大的现有生产设施。
发明内容
本发明的一个目的涉及一种重组无细胞反应组分；包含体外转录/翻译机制、氨基酸、核苷酸、代谢组分所需的所有无细胞反应组分的反应混合物，这些组分可提供能量，对于蛋白合成必不可少。在一个优选实施例中，本文所确定的酶可来源于不同的嗜热菌，而传统无细胞系统的组分则来源于大肠杆菌或其他真核系统(如酵母)。这种嗜热源策略在体外翻译的所有步骤(tRNA加载、核糖体肽生物合成)中提供更高的稳定性，还可以改进重组表达系统的性能，并延长其运行时间。
发明人的嗜热源策略可产生一种重组无细胞表达系统，这种系统对pH和盐浓度变化的灵敏度较低，并且受ATP水解而导致磷酸盐浓度增加的影响较小。这种嗜热源策略的另一个好处在于，使创新性无细胞表达系统能够利用不同的tRNA集合(通过嗜热氨酰-tRNA合成酶识别)，从而在无细胞系统中首次实现全密码子覆盖。
本专利的另一个目的可包括一种重组无细胞表达系统，其中，反应混合物包含生物化合物、蛋白、酶、生物仿制药体外生物合成或小分子化学修饰所需的所有无细胞反应组分。
本发明的另一个目的可包括一种连续流动式生物反应器系统的各种方法、系统和装置，可用于对疫苗、生物制剂、蛋白、酶、生物仿制药进行体外转录、体外翻译和体外生物合成以及在连续流动操作中使用酶对小分子进行生物合成或化学修饰。
本发明的另一个目的可包括一个或多个分离核苷酸编码序列，这些序列可形成重组无细胞表达反应混合物的一部分。在一个优选实施例中，一个或多个核苷酸编码序列可来自嗜热菌或其他细菌。在一个优选实施例中，核苷酸编码序列可包括但不限于：起始因子核苷酸编码序列、延伸因子核苷酸编码序列、释放因子核苷酸编码序列、核糖体循环因子核苷酸编码序列、氨酰-tRNA合成酶核苷酸编码序列和甲硫氨酰-tRNA转甲酰基酶核苷酸编码序列。其他核苷酸编码序列可包括RNA聚合酶核苷酸编码序列以及在本文所纳入的参考文件第PCT/US201 8/012121号PCT申请(“121申请”)中确定的核苷酸编码序列，所述申请涉及本文所纳入的无机多磷酸盐能量再生系统。
本发明的另一个目的可包括产生包含一个或多个分离核苷酸编码序列的表达载体，所述分离核苷酸编码序列可连接用于转化细菌细胞的启动子序列。在某些实施例中，可对核苷酸编码序列进行优化，以在所选细菌中进行表达。
本发明的另一个目的可包括表达本文所确定的核苷酸编码序列，产生可进一步分离并包含在重组无细胞表达反应混合物中的蛋白。在一个优选实施例中，所表达的蛋白可包括但不限于：起始因子蛋白、延伸因子蛋白、释放因子蛋白、核糖体循环因子蛋白、氨酰-tRNA合成酶蛋白和甲硫氨酰-tRNA转甲酰基酶蛋白。其他核苷酸编码序列可包括RNA聚合酶蛋白以及在“121申请”中确定的蛋白和化合物，所述申请涉及本文所纳入的无机多磷酸盐能量再生系统。
本发明的另一个目的可包括连续流动式重组无细胞表达装置。在本优选实施例中，此类连续流动式重组无细胞表达装置可包括将中空纤维和中空纤维生物反应器作为交换介质，对生物制剂、蛋白、酶、生物仿制药进行体外转录、体外翻译和体外生物合成以及在连续流动操作中使用酶对小分子进行生物合成或化学修饰。
本发明的其他目的可包括以下一项或多项优选实施例：
1.一种重组无细胞表达系统，包含：
-一种至少包含以下组分的核心重组蛋白混合物：
-多个起始因子(IF)；
-多个延伸因子(EF)；
-多个肽释放因子(RF)；
-至少一个核糖体循环因子(RRF)；
-多个氨酰-tRNA合成酶(RS)；以及
-至少一个甲硫氨酰-tRNA转甲酰基酶(MTF)；
-至少一个核酸合成模板；
-一种包含体外大分子合成所需的无细胞反应组分的反应混合物；以及
-其中，上述组分位于一个可进行大分子无细胞表达的生物反应器中。
2.根据实施例1所述的系统，其中，所述核心重组蛋白混合物包括一种由细菌衍生的核心重组蛋白混合物。
3.根据实施例2所述的系统，其中，所述由细菌衍生的核心重组蛋白混合物包括一种核心重组蛋白混合物，其中，至少一种组分由嗜热菌衍生而来。
4.根据实施例2和3中任一项所述的系统，其中，所述嗜热菌包括芽孢杆菌嗜热菌或地芽孢杆菌嗜热菌。
5.根据实施例4所述的系统，其中，从包括以下几项的一组中选择所述地芽孢杆菌嗜热菌：地表地芽孢杆菌和嗜热脂肪地芽孢杆菌。
6.根据实施例1所述的系统，其中，所述由细菌衍生的核心重组蛋白混合物包括一种核心重组蛋白混合物，其中，至少一种组分由非嗜热菌或非嗜热菌和嗜热菌的组合衍生而来。
7.根据实施例6所述的系统，其中，所述非嗜热菌包括大肠杆菌。
8.根据实施例1所述的系统，其中，所述多个起始因子(IF)包括由嗜热菌衍生的多个起始因子。
9.根据实施例1和8中任一项所述的系统，其中，所述由嗜热菌衍生的多个起始因子包括IF1、IF2、IF3或其任何片段或变体。
10.根据实施例1、8和9中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个起始因子：SEQ ID NO.2、4、6、70、72和74，或序列一致性至少达90％的一个序列。
11.根据实施例1所述的系统，其中，所述多个延伸因子(EF)包括由嗜热菌衍生的多个延伸因子。
12.根据实施例1和11中任一项所述的系统，其中，所述由嗜热菌衍生的多个延伸因子包括EF-G、EF-Tu、EF-Ts、EF-4、EF-P或其任何片段或变体。
13.根据实施例1、11和12中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个延伸因子：SEQ ID NO.8、10、12、14、16、76、78、80、82和84，或序列一致性至少达90％的一个序列。
14.根据实施例1所述的系统，其中，所述多个肽释放因子(RF)包括由嗜热菌或芽孢杆菌衍生的多个肽释放因子。
15.根据实施例1和14中任一项所述的系统，其中，所述由嗜热菌衍生的多个肽释放因子包括RF1、RF2和RF3或其任何片段或变体。
16.根据实施例1、14和15中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个肽释放因子：SEQ ID NO.18、20、22、86和88，或序列一致性至少达90％的一个序列。
17.根据实施例1所述的系统，其中，所述核糖体循环因子(RRF)包括由嗜热菌衍生的核糖体循环因子。
18.根据实施例1和17中任一项所述的系统，其中，所述核糖体循环因子由地芽孢杆菌衍生而来。
19.根据实施例1、17和18中任一项所述的系统，其中，所述核糖体循环因子包括包含以下氨基酸序列的核糖体循环因子：SEQ ID NO.14和90，或序列一致性至少达90％的一个序列。
20.根据实施例1所述的系统，其中，所述多个氨酰-tRNA合成酶(RS)包括由嗜热菌或大肠杆菌衍生的多个氨酰-tRNA合成酶。
21.根据实施例1和20中任一项所述的系统，其中，所述多个氨酰-tRNA合成酶包括AlaRS、ArgRS、AsnRS、AspRS、CysRS、GlnRS、GluRS、GlyRS、HisRS、IleRS、LeuRS、LysRS、MetRS、PheRS(a)、PheRS(b)、ProRS、SerRS、ThrRS、TrpRS、TyrRS和ValRS或其任何片段或变体。
22.根据实施例1、20和21中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个氨酰-tRNA合成酶：SEQ ID NO.26、28、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、94、96、98、100、102、104、106、108、110、112、114、116、118、120、122、124、126、128和130，或序列一致性至少达90％的一个序列。
23.根据实施例1所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶(MTF)包括由嗜热菌衍生的甲硫氨酰-tRNA转甲酰基酶。
24.根据实施例1和23中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶由地芽孢杆菌衍生而来。
25.根据实施例1、23和24中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶包括包含以下氨基酸序列的甲硫氨酰-tRNA转甲酰基酶：SEQ ID NO.68和132，或序列一致性至少达90％的一个序列。
26.根据实施例1所述的系统，其中，所述核酸合成模板包括DNA模板。
27.根据实施例26所述的系统，其中，所述DNA模板包括线性DNA模板，其包含：
-至少一个可连接启动子的靶序列，其中，所述靶序列可进行密码子优化；
-至少一个核糖体结合位点(RBS)；
-至少一个表达产物酶切位点；以及
-至少一个标签。
28.根据实施例1所述的系统，其中，所述核酸合成模板包括RNA模板。
29.根据实施例1所述的系统，其中，所述反应混合物包含以下一种或多种组分：
-.一定量的核糖体，以及一定量的由嗜热菌衍生的核糖体(可选)；
-.一定量的RNase抑制剂；
-.一定量的RNA聚合酶；
-.一定量的tRNA，以及一定量的由嗜热菌衍生的tRNA(可选)；
-.一种缓冲液；以及
-.一定量的氨基酸。
30.根据实施例29所述的系统，其中，所述反应混合物进一步包含以下一种或多种组分：
-Tris乙酸盐；
-Mg(OAc)2；
-K+谷氨酸；
-氨基乙酸盐；
-NaCl；
-KC1；
-MgCk；
-DTT；
-辛基-b-糖苷；
-NAD；
-NADP；
-山梨糖醇；
-FADH；
-CoA；
-PLP；以及
-SAM。
31.根据实施例1和29中任一项所述的系统，进一步包含一个能量源：
32.根据实施例32所述的系统，其中，所述能量源包括一定量的核苷三磷酸(NTP)。
33.根据实施例32所述的系统，其中，所述核苷三磷酸包括从包含以下几项的一组中选择的一种或多种核苷三磷酸：腺嘌呤三磷酸(ATP)、鸟苷三磷酸(GTP)、尿苷三磷酸(UTP)和胞苷三磷酸(CTP)。
34.根据实施例31、32和33中任一项所述的系统，其中，所述能量源包括一个无机多磷酸盐能量再生系统。
35.根据实施例34所述的系统，其中，所述无机多磷酸盐能量再生系统包括：
-一个细胞三磷酸腺苷(ATP)能量再生系统，包含：
-一定量的腺苷激酶(GstAdK)；
-一定量的多磷酸盐激酶(Taq PPK)；
-一定量的无机多磷酸盐(PPi)；以及
-一定量的一磷酸腺苷(AMP)；
-其中，所述AdK酶和PPK酶发挥协同作用，从PPi和AMP中再生细胞能量。
36.根据实施例1所述的系统，其中，所述生物反应器包括一个连续流动式生物反应器。
37.一种重组无细胞表达反应混合物，包含：
-多个起始因子(IF)；
-多个延伸因子(EF)；
-多个释放因子(RF)；
-至少一个核糖体循环因子(RRF)；
-多个氨酰-tRNA合成酶(RS)；以及
-至少一个甲硫氨酰-tRNA转甲酰基酶(MTF)；
38.根据实施例37所述的系统，其中，所述多个起始因子(IF)包括由嗜热菌衍生的多个起始因子。
39.根据实施例37和38中任一项所述的系统，其中，所述由嗜热菌衍生的多个起始因子包括IF1、IF2、IF3或其任何片段或变体。
40.根据实施例37、38和39中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个起始因子：SEQ ID NO.2、4、6、70、72和74，或序列一致性至少达90％的一个序列。
41.根据实施例37所述的系统，其中，所述多个延伸因子(EF)包括由嗜热菌衍生的多个延伸因子。
42.根据实施例37和41中任一项所述的系统，其中，所述由嗜热菌衍生的多个延伸因子包括EF-G、EF-Tu、EF-Ts、EF-4、EF-P或其任何片段或变体。
43.根据实施例37、41和42中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个延伸因子：SEQ ID NO.8、10、12、14、16、76、78、80、82和84，或序列一致性至少达90％的一个序列。
44.根据实施例37所述的系统，其中，所述多个肽释放因子(RF)包括由嗜热菌或芽孢杆菌衍生的多个释放因子。
45.根据实施例37和44中任一项所述的系统，其中，所述多个肽释放因子包括RF1、RF2和RF3或其任何片段或变体。
46.根据实施例37、44和45中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个肽释放因子：SEQ ID NO.18、20、22、86和88，或序列一致性至少达90％的一个序列。
47.根据实施例37所述的系统，其中，所述核糖体循环因子(RRF)包括由嗜热菌衍生的核糖体循环因子。
48.根据实施例37和47中任一项所述的系统，其中，所述核糖体循环因子由地芽孢杆菌衍生而来。
49.根据实施例37、47和48中任一项所述的系统，其中，所述核糖体循环因子包括包含以下氨基酸序列的核糖体循环因子：SEQ ID NO.14和90，或序列一致性至少达90％的一个序列。
50.根据实施例37所述的系统，其中，所述多个氨酰-tRNA合成酶(RS)包括多个氨酰-tRNA合成酶，其中，至少一个氨酰-tRNA合成酶由嗜热菌衍生而来。
51.根据实施例37和50中任一项所述的系统，其中，所述多个氨酰-tRNA合成酶包括AlaRS、ArgRS、AsnRS、AspRS、CysRS、GlnRS、GluRS、GlyRS、HisRS、IleRS、LeuRS、LysRS、MetRS、PheRS(a)、PheRS(b)、ProRS、SerRS、ThrRS、TrpRS、TyrRS和ValRS或其任何片段或变体。
52.根据实施例37、50和51中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个氨酰-tRNA合成酶：SEQ ID NO.26、28、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、94、96、98、100、102、104、106、108、110、112、114、116、118、120、122、124、126、128和130，或序列一致性至少达90％的一个序列。
53.根据实施例37所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶(MTF)包括由嗜热菌衍生的甲硫氨酰-tRNA转甲酰基酶。
54.根据实施例37和53中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶由地芽孢杆菌衍生而来。
55.根据实施例37、53和54中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶包括包含以下氨基酸序列的甲硫氨酰-tRNA转甲酰基酶：SEQ ID NO.68和132，或序列一致性至少达90％的一个序列。
56.一种分离核苷酸，包括从包含以下几项的一组中选择的一种核苷酸：
-SEQ ID NO.1、3、5、69、71和73；
-SEQ ID NO.7、9、11、13、15、75、77、79、81和83；
-SEQ ID NO.17、19、21、85和87；
-SEQ ID NO.23和89；以及
-SEQ ID NO.25、27、29、31、33、35、37、39、41、43、45、47、49、51、53、55、57、59、61、63、65、67、91、93、95、97、99、101、103、105、107、109、111、113、115、117、119、121、123、125、127、129和131。
57.一种表达载体，包含至少一个实施例56所述的核苷酸序列(可连接启动子)。
58.一种由实施例57所述的一种表达载体转化的细菌。
59.根据实施例58所述的转化细菌，其中，所述细菌包括大肠杆菌。
60.一种肽，包含从包括以下几项的一组中选择的一个氨基酸序列：
-SEQ ID NO.2、4、6、70、72和74；
-SEQ ID NO.8、10、12、14、16、76、78、80、82和84；
-SEQ ID NO.18、20、22、86、88；
-SEQ ID NO.14和90；
-SEQ ID NO.26、28、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、94、96，SEQ ID NO.98、100、102、104、106、108、110、112、114、116、118、120、122、124、126、128和130；以及
-SEQ ID NO.68和132，或其任何片段或变体。
61.一种无细胞表达系统，其使用实施例60所述的至少一种肽。
通过下文所述的具体实施方式、附图和权利要求，创新性技术的其他目的显而易见。
附图说明
附图纳入并构成说明书的一部分，显示了本发明的一个或多个实施例，并结合说明解释了创新性技术的某些方面。附图仅旨在说明本发明的一个或多个优选实施例，不应视为对本发明的限制。
图1：显示了合成酶：AlaRS、ArgRS、AsnRS、AspRS、CysRS、GlnRS(Ec)、GluRS、GlyRS、HisRS和IleRS以及无tRNA对照品的氨酰-tRNA合成酶动力学活性检测结果。
图2：显示了合成酶：LeuRS、LysRS、MetRS、PheRS、ProRS、SerRS、ThrRS、TrpRS、TyrRS和ValRS以及无tRNA对照品的氨酰-tRNA合成酶动力学活性检测结果。
图3A：显示了利用示例性tRNA(来自大肠杆菌)的氨酰-tRNA合成酶活性检测的结果。
图3B：显示了利用tRNA(来自示例性嗜热菌嗜热脂肪地芽孢杆菌)的氨酰-tRNA合成酶活性检测的结果。
图4：显示了利用本文所述的重组无细胞表达系统生产绿色荧光蛋白(muGFP，SEQID NO.134)无细胞表达产物的情况。
图5：在其一个实施例中用于无细胞生产和连续交换的中空纤维反应器示意图。
图6A-B：在其一个实施例中用于无细胞生产和连续交换的中空纤维反应器图像。
图7：使用pET151/D-TOPO载体选择合成基因，此类合成基因向所表达的蛋白添加N端标签。在此载体中表达的所有基因均从蛋白序列逆翻译为DNA，并进行密码子优化，以在大肠杆菌中进行表达。在下文确定的特异性序列中，可省略N端标签。
图8：使用pET24a(+)载体选择合成基因，此类合成基因向所表达的蛋白添加C端6XHis标签。在此载体中表达的所有基因从蛋白序列逆翻译为DNA，并进行密码子优化，以在大肠杆菌中进行表达。在下文确定的特异性序列中，可省略C端标签。
图9：设计并使用pNAT载体选择克隆和/或合成基因，此类克隆和/或合成基因向所表达的蛋白添加N端FLAG标签和/或C端6X His标签。在此载体中表达的所有基因从蛋白序列逆翻译为DNA，并进行密码子优化，以在大肠杆菌中进行表达。在下文确定的特异性序列中，可省略标签。
图10：设计并使用pNAT 2.0载体选择克隆和/或合成基因，此类克隆和/或合成基因向所表达的蛋白添加N端或C端6X His标签。在此载体中表达的所有基因均从蛋白序列逆翻译为DNA，并进行密码子优化，以在大肠杆菌中进行表达。在下文确定的特异性序列中，可省略标签。
图11：显示了经纯化的氨酰-tRNA合成酶(aaRS)AlaRS、ArgRS、AsnRS、AspRS、CysRS、GlnRS(Ec)、GluRS、GlyRS、HisRS、IleRS和LeuRS的SDS-PAGE结果。
图12：显示了经纯化的氨酰-tRNA合成酶(aaRS)LysRS、MetRS、PheBRS、ProRS、SerRS、ThrRS、TrpRS、TyrRS和ValRS以及经纯化的甲硫氨酰-tRNA转甲酰基酶(MTF)的SDS-PAGE结果。
图13：显示了经纯化的翻译因子IF-1、IF-2、IF-3、EF-G、EF-Ts、EF-Tu、EF-P、RF-1、RF-2、RF-3和RRF的SDS-PAGE结果。
图14：显示了经纯化的翻译因子EF-4的SDS-PAGE结果。
图15：显示了利用本文所述的重组无细胞表达系统实时生产荧光蛋白(muGFP；SEQID NO.134)产物的情况。
图16：显示了在逆纯化后但未过滤掉核糖体时使用无细胞蛋白表达反应的抗FLAG抗体执行蛋白免疫印迹法的情况。显示了利用本文所述的重组无细胞表达系统对蛋白无细胞表达产物(特别是不稳定增强型绿色荧光蛋白(deGFP，SEQ ID NO.135))进行特异性检测的情况。
图17：(A)显示了利用示例性tRNA(来自大肠杆菌)的三种独立氨酰-tRNA合成酶AMP生产活性检测的结果。(B)显示了AMP标准曲线。
具体实施方式
本发明特别适用于在无细胞环境中按需生产多肽等治疗性大分子，而所述治疗性大分子可直接输送给患者。因此，主要结合治疗性蛋白的生产对本发明进行了说明。但是，本发明也可用于生产任何类型的蛋白，包括有毒蛋白、含放射性标记氨基酸和非天然氨基酸的蛋白等。此外，本发明特别适用于通过无细胞表达按需生产蛋白，因此，主要在无细胞蛋白表达的情况下对本发明进行了说明。
本发明包括可以不同方式组合的各个方面。以下说明列出了各种要素，并对本发明的一些实施例进行了说明。这些要素与初始实施例一起列出；但是，应理解，它们可以任何方式和数量组合，以创建其他实施例。各种说明示例和优选实施例不应视为将本发明限制为仅明确说明的系统、技术和应用。此外，应理解，本说明旨在支持并包含各种实施例、系统、技术、方法、装置和应用的说明和权利要求以及任意数量的公开要素、每个单独要素和本申请或任何后续申请中所有要素的任何及所有不同的排列和组合。
本文所述的创新性技术可包括一种新型重组无细胞表达系统。在一个优选实施例中，本发明可包括产生一种反应混合物，所述反应混合物包含可促进体外表达活性的多个核心部分。示例性核心蛋白可包括以下几项：
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一个或多个起始因子(IF)。起始因子可在肽合成的过程中形成起始复合物。例如，在某些实施例中，IF1、IF2和IF3可用作反应混合物中的起始因子。例如，IF3促进将核糖体解离成30S和50S亚基(即，开始翻译通常所需的步骤)，并在形成起始复合物的步骤中，阻止将除甲酰甲硫氨酰-tRNA之外的tRNA插入P位置。IF2结合甲酰甲硫氨酰-tRNA，并将甲酰甲硫氨酰-tRNA转移到30S亚基的P位置，从而形成起始复合物。IFl可增强IF2和IF3的功能。在本发明中，优选使用来自一种或多种细菌的起始因子，更优选嗜热菌，例如，从菌群芽孢杆菌和/或地芽孢杆菌(如地表地芽孢杆菌或嗜热脂肪地芽孢杆菌)中获得的起始因子。可从包括以下几项的一组中选择本发明一个或多个IF的示例性氨基酸序列：
IF1(SEQ ID NO.2和70)
IF2(SEQ ID NO.4和72)
IF3(SEQ ID NO.6和74)
因此，在本发明的一个实施例中，上述一个或多个氨基酸序列包含至少一个IF，所述IF包含一个氨基酸序列，而此类氨基酸序列由SEQ ID NO.1-2、4、6、69-70、72和74氨基酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个IF的至少一个编码区编码的蛋白片段或其变体可通常包含一个氨基酸序列，此类氨基酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的氨基酸序列优选本文公开的SEQ ID NO.1-2、4、6、69-70、72和74。
在本发明中，优选使用在一种或多种细菌中表达和/或分离的起始因子，更优选表达高水平蛋白的细菌，如大肠杆菌。可从包括以下几项的一组中选择本发明一个或多个IF的示例性核苷酸序列：
IF1(SEQ ID NO.1和69)
IF2(SEQ ID NO.3和71)
IF3(SEQ ID NO.5和73)
显而易见，可对核苷酸序列进行密码子优化，以在一种或多种细菌或其他蛋白表达系统(如酵母等)中进行表达。例如，在本实施例中，已对示例性核苷酸序列SEQ ID NO.1、3和5进行密码子优化，以在大肠杆菌中进行表达。
因此，在本发明的一个实施例中，上述一个或多个核苷酸序列包含至少一个编码至少一个IF的编码区，所述IF包含一个核苷酸序列，而此类核苷酸序列由SEQ ID NO.1、3、5、69、71和73核苷酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个IF的至少一个编码区编码的蛋白片段或其变体可通常包含一个核苷酸序列，此类核苷酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的核苷酸序列优选本文公开的SEQ ID NO.1、3、5、69、71和73。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一个或多个延伸因子。EF-Tu等延伸因子可分为2类，即GTP型和GDP型。GTP型EF-Tu结合氨酰-tRNA，并将其转移到核糖体的A位置。当从核糖体中释放EF-Tu时，GTP水解成GDP。另一个延伸因子EF-Ts结合GDP型EF-Tu，并促进将其转化为GTP型。在肽链延长的过程中，另一个延伸因子EF-G可在肽键形成后促进易位。在本发明中，优选使用来自细菌的EF，更优选嗜热菌，例如，从菌群芽孢杆菌和/或地芽孢杆菌(如地表地芽孢杆菌或嗜热脂肪地芽孢杆菌)中获得的EF。可从包括以下几项的一组中选择本发明一个或多个EF的示例性氨基酸序列：
EF-G(SEQ ID NO.8和76)
EF-Tu(SEQ ID NO.10和78)
EF-Ts(SEQ ID NO.12和80)
EF-4(SEQ ID NO.14和82)
EF-P(SEQ ID NO.16和84)
因此，在本发明的一个实施例中，上述一个或多个氨基酸序列包含至少一个EF，所述EF包含一个氨基酸序列，而此类氨基酸序列由SEQ ID NO.8、10、12、14、16、76、78、80、82和84氨基酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个EF的至少一个编码区编码的蛋白片段或其变体可通常包含一个氨基酸序列，此类氨基酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的氨基酸序列优选本文公开的SEQ ID NO.8、10、12、14、16、76、78、80、82和84。
在本发明中，优选使用在一种或多种细菌中表达和/或分离的EF，更优选表达高水平蛋白的细菌，如大肠杆菌。可从包括以下几项的一组中选择本发明一个或多个EF的示例性核苷酸序列：
EF-G(SEQ ID NO.7和75)
EF-Tu(SEQ ID NO.9和77)
EF-Ts(SEQ ID NO.11和79)
EF-4(SEQ ID NO.13和81)
EF-P(SEQ ID NO.15和83)
显而易见，可对核苷酸序列进行密码子优化，以在一种或多种细菌或其他蛋白表达系统(如酵母等)中进行表达。例如，在本实施例中，已对示例性核苷酸序列SEQ ID NO.7、9、11、13和15进行密码子优化，以在大肠杆菌中进行表达。
因此，在本发明的一个实施例中，上述一个或多个核苷酸序列包含至少一个编码至少一个EF的编码区，所述EF包含一个核苷酸序列，而此类核苷酸序列由SEQ ID NO.7、9、11、13、15、75、77、79和83核苷酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个EF的至少一个编码区编码的蛋白片段或其变体可通常包含一个核苷酸序列，此类核苷酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的核苷酸序列优选本文公开的SEQ ID NO.7、9、11、13、15、75、77、79和83。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一个或多个肽释放因子(RF)。RF可负责终止蛋白合成，释放经翻译的肽链，并循环核糖体，以开始后续mRNA翻译。在无释放因子的反应系统中合成蛋白时，反应在终止密码子前停止，因此很容易形成由核糖体、肽和mRNA组成的稳定三元复合物(多核糖体展示)。当终止密码子(UAA、UAG或UGA)位于核糖体的A位置时，释放因子RF1和RF2可进入A位置，并促进P位置处肽基tRNA的肽链解离。RF1识别终止密码子中的UAA和UAG，而RF2则识别UAA和UGA。在通过RF1和RF2解离肽链后，另一个终止因子RF3促进从核糖体中解离RF1和RF2。
在本发明中，优选使用来自细菌的RF，更优选嗜热菌，例如，从菌群芽孢杆菌和/或地芽孢杆菌(如地表地芽孢杆菌或嗜热脂肪地芽孢杆菌)中获得的RF。可从包括以下几项的一组中选择本发明一个或多个RF的示例性氨基酸序列：
RF1(SEQ ID NO.18和86)
RF2(SEQ ID NO.20和88)
RF3(SEQ ID NO.22)
因此，在本发明的一个实施例中，上述一个或多个氨基酸序列包含至少一个RF，所述RF包含一个氨基酸序列，而此类氨基酸序列由SEQ ID NO.18、20、22、86和88氨基酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个RF的至少一个编码区编码的蛋白片段或其变体可通常包含一个氨基酸序列，此类氨基酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的氨基酸序列优选本文公开的SEQ ID NO.18、20、22、86和88。
在本发明中，优选使用在一种或多种细菌中表达和/或分离的RF，更优选表达高水平蛋白的细菌，如大肠杆菌。可从包括以下几项的一组中选择本发明一个或多个RF的示例性核苷酸序列：
RF1(SEQ ID NO.17和85)
RF2(SEQ ID NO.19和87)
RF3(SEQ ID NO.21)
显而易见，可对核苷酸序列进行密码子优化，以在一种或多种细菌或其他蛋白表达系统(如酵母等)中进行表达。例如，在本实施例中，已对示例性核苷酸序列SEQ IDNO.17、19和21进行密码子优化，以在大肠杆菌中进行表达。
因此，在本发明的一个实施例中，上述一个或多个核苷酸序列包含至少一个编码至少一个RF的编码区，所述RF包含一个核苷酸序列，而此类核苷酸序列由SEQ ID NO.17、19、21、85和87核苷酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个RF的至少一个编码区编码的蛋白片段或其变体可通常包含一个核苷酸序列，此类核苷酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的核苷酸序列优选本文公开的SEQ ID NO.17、19、21、85和87。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一个或多个核糖体循环因子(RRF)，在蛋白合成及循环核糖体以进行后续蛋白合成后，所述核糖体循环因子可促进P位置剩余tRNA的解离。在本发明中，优选使用来自细菌的RRF，更优选嗜热菌，例如，从菌群芽孢杆菌和/或地芽孢杆菌(如地表地芽孢杆菌或嗜热脂肪地芽孢杆菌)中获得的RRF。可从包括以下几项的一组中选择本发明一个或多个RRF的示例性氨基酸序列：
RRF(SEQ ID NO.24和90)
因此，在本发明的一个实施例中，上述一个或多个氨基酸序列包含至少一个RRF，所述RRF包含一个氨基酸序列，而此类氨基酸序列由SEQ ID NO.23和90氨基酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个RRF的至少一个编码区编码的蛋白片段或其变体可通常包含一个氨基酸序列，此类氨基酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的氨基酸序列优选本文公开的SEQ ID NO.23和90。
在本发明中，优选使用在一种或多种细菌中表达和/或分离的RRF，更优选表达高水平蛋白的细菌，如大肠杆菌。可从包括以下几项的一组中选择本发明一个或多个RRF的示例性核苷酸序列：
RRF(SEQ ID NO.23和89)
显而易见，可对核苷酸序列进行密码子优化，以在一种或多种细菌或其他蛋白表达系统(如酵母等)中进行表达。例如，在本实施例中，已对示例性核苷酸序列SEQ ID NO.23进行密码子优化，以在大肠杆菌中进行表达。
因此，在本发明的一个实施例中，上述一个或多个核苷酸序列包含至少一个编码至少一个RF的编码区，所述RF包含一个核苷酸序列，而此类核苷酸序列由SEQ ID NO.23和89核苷酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个RF的至少一个编码区编码的蛋白片段或其变体可通常包含一个核苷酸序列，此类核苷酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的核苷酸序列优选本文公开的SEQ IDNO.23和89。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一个或多个氨酰-tRNA合成酶(RS)。存在ATP时，通过氨酰-tRNA合成酶，氨基酸可与tRNA共价结合，从而合成氨酰-tRNA。在本发明中，优选使用嗜热菌来源的氨酰-tRNA合成酶，例如，从菌群芽孢杆菌和/或地芽孢杆菌中获得的氨酰-tRNA合成酶，更具体地说，从菌种嗜热脂肪地芽孢杆菌中获得的氨酰-tRNA合成酶。其他实施例可包括使用来自非嗜热菌(如大肠杆菌)的氨酰-tRNA合成酶，这种应用与嗜热菌来源的氨酰-tRNA合成酶结合。从包括以下几项的一组中选择氨酰-tRNA合成酶的示例性核苷酸序列和氨基酸序列：
AlaRS (SEQ ID NO.26和SEQ ID NO.92)
ArgRS (SEQ ID NO.28和SEQ ID NO.94)
AsnRS (SEQ ID NO.30和SEQ ID NO.96)
AspRS (SEQ ID NO.32和SEQ ID NO.98)
CysRS (SEQ ID NO.34和SEQ ID NO.100)
GlnRS(Ec) (SEQ ID NO.36)
GluRS (SEQ ID NO.38和SEQ ID NO.102)
GlyRS (SEQ ID NO.40和SEQ ID NO.104)
HisRS (SEQ ID NO.42和SEQ ID NO.106)
IleRS (SEQ ID NO.44和SEQ ID NO.108)
LeuRS (SEQ ID NO.46和SEQ ID NO.110)
LysRS (SEQ ID NO.48和SEQ ID NO.112)
MetRS (SEQ ID NO.50和SEQ ID NO.114)
PheRS(a) (SEQ ID NO.52和SEQ ID NO.116)
PheRS(b) (SEQ ID NO.54和SEQ ID NO.118)
ProRS (SEQ ID NO.56和SEQ ID NO.120)
SerRS (SEQ ID NO.58和SEQ ID NO.122)
ThrRS (SEQ ID NO.60和SEQ ID NO.124)
TrpRS (SEQ ID NO.62和SEQ ID NO.126)
TyrRS (SEQ ID NO.64和SEQ ID NO.128)
ValRS (SEQ ID NO.66和SEQ ID NO.130)
因此，在本发明的一个实施例中，上述一个或多个氨基酸序列包含至少一个RS，所述RS包含一个氨基酸序列，而此类氨基酸序列由SEQ ID NO.26、28、30、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、92、94、96、98、100、102、104、106、108、110、112、114、116、118、120、122、134、126、128和130氨基酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个RS的至少一个编码区编码的蛋白片段或其变体可通常包含一个氨基酸序列，此类氨基酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的氨基酸序列优选本文公开的SEQ IDNO.26、28、30、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、92、94、96、98、100、102、104、106、108、110、112、114、116、118、120、122、134、126、128和130。
在本发明中，优选使用在一种或多种细菌中表达和/或分离的RS，更优选表达高水平蛋白的细菌，如大肠杆菌。可从包括以下几项的一组中选择本发明一个或多个RS的示例性核苷酸序列：
AlaRS (SEQ ID NO.25和SEQ ID NO.91)
ArgRS (SEQ ID NO.27和SEQ ID NO.93)
AsnRS (SEQ ID NO.29和SEQ ID NO.95)
AspRS (SEQ ID NO.31和SEQ ID NO.97)
CysRS (SEQ ID NO.33和SEQ ID NO.99)
GlnRS(Ec) (SEQ ID NO.35)
GluRS (SEQ ID NO.37和SEQ ID NO.101)
GlyRS (SEQ ID NO.39和SEQ ID NO.103)
HisRS (SEQ ID NO.41和SEQ ID NO.105)
IleRS (SEQ ID NO.43和SEQ ID NO.107)
LeuRS (SEQ ID NO.45和SEQ ID NO.109)
LysRS (SEQ ID NO.47和SEQ ID NO.111)
MetRS (SEQ ID NO.49和SEQ ID NO.113)
PheRS(a) (SEQ ID NO.51和SEQ ID NO.115)
PheRS(b) (SEQ ID NO.53和SEQ ID NO.117)
ProRS (SEQ ID NO.55和SEQ ID NO.119)
SerRS (SEQ ID NO.57和SEQ ID NO.121)
ThrRS (SEQ ID NO.59和SEQ ID NO.123)
TrpRS (SEQ ID NO.61和SEQ ID NO.125)
TyrRS (SEQ ID NO.63和SEQ ID NO.127)
ValRS (SEQ ID NO.65和SEQ ID NO.129)
显而易见，可对核苷酸序列进行密码子优化，以在一种或多种细菌或其他蛋白表达系统(如酵母等)中进行表达。例如，在本实施例中，已对示例性核苷酸序列SEQ IDNO.25、27、29、31、33、35、37、39、41、43、45、47、49、51、53、55、57、59、61、63和65进行密码子优化，以在大肠杆菌中进行表达。
因此，在本发明的一个实施例中，上述一个或多个核苷酸序列包含至少一个编码至少一个RS的编码区，所述RS包含一个核苷酸序列，而此类核苷酸序列由SEQ ID NO.25、27、29、31、33、35、37、39、41、43、45、47、49、51、53、55、57、59、61、63、65、91、93、95、97、99、101、103、105、107、109、111、113、115、117、119、121、123、125、127和129核苷酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个RS的至少一个编码区编码的蛋白片段或其变体可通常包含一个核苷酸序列，此类核苷酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的核苷酸序列优选本文公开的SEQ ID NO.25、27、29、31、33、35、37、39、41、43、45、47、49、51、53、55、57、59、61、63、65、91、93、95、97、99、101、103、105、107、109、111、113、115、117、119、121、123、125、127和129。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一个甲硫氨酰-tRNA转甲酰基酶(MTF)。N-甲酰甲硫氨酸携带一个甲酰基(连接到蛋氨酸末端的氨基上)，在原核蛋白合成系统中作为起始氨基酸。甲酰基通过MTF连接到甲硫氨酰-tRNA中的蛋氨酸上。也就是说，MTF将Nlv-甲酰四氢叶酸中的甲酰基转移到甲硫氨酰-tRNA(对应于起始密码子)的N端，从而提供甲酰甲硫氨酰-tRNA。由此连接的甲酰基被IF2识别，并作为蛋白合成的起始信号。在本发明中，优选使用来自细菌的MTF，更优选嗜热菌，例如，从菌群芽孢杆菌和/或地芽孢杆菌(如地表地芽孢杆菌或嗜热脂肪地芽孢杆菌)中获得的MTF。可从包括以下几项的一组中选择本发明一个或多个MTF的示例性氨基酸序列：
MTF(SEQ ID NO.68和132)
因此，在本发明的一个实施例中，上述一个或多个氨基酸序列包含至少一个MTF，所述MTF包含一个氨基酸序列，而此类氨基酸序列由SEQ ID NO.68和132氨基酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个MTF的至少一个编码区编码的蛋白片段或其变体可通常包含一个氨基酸序列，此类氨基酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的氨基酸序列优选本文公开的SEQ ID NO.68和132。
在本发明中，优选使用在一种或多种细菌中表达和/或分离的MTF，更优选表达高水平蛋白的细菌，如大肠杆菌。可从包括以下几项的一组中选择本发明一个或多个MTF的示例性核苷酸序列：
MTF(SEQ ID NO.67和131)
显而易见，可对核苷酸序列进行密码子优化，以在一种或多种细菌或其他蛋白表达系统(如酵母等)中进行表达。例如，在本实施例中，已对示例性核苷酸序列SEQ ID NO.67进行密码子优化，以在大肠杆菌中进行表达。
因此，在本发明的一个实施例中，上述一个或多个核苷酸序列包含至少一个编码至少一个MTF的编码区，所述MTF包含一个核苷酸序列，而此类核苷酸序列由SEQ ID NO.67和131核苷酸序列或其一个片段或变体编码。在这种情况下，由本发明所述一个或多个MTF的至少一个编码区编码的蛋白片段或其变体可通常包含一个核苷酸序列，此类核苷酸序列的序列一致性为至少5％、10％、20％、30％、40％、50％、60％、70％、80％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％，优选至少70％，更优选至少80％，甚至更优选至少85％，甚至更优选至少90％，最优选至少95％或甚至97％，相应天然全长蛋白或其变体的核苷酸序列优选本文公开的SEQ ID NO.67和131。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一定量的核糖体。核糖体是一种合成肽的颗粒，可与mRNA结合，并将氨酰-tRNA协调到A位置，将甲酰甲硫氨酰-tRNA或肽基tRNA协调到P位置，从而形成肽键。在本发明中，可使用各种来源的任何核糖体，但是，在一个优选实施例中，可从用于重组无细胞表达系统的嗜热菌中分离核糖体，优选从嗜热菌的细胞裂解物中分离核糖体，如从菌群芽孢杆菌和/或地芽孢杆菌(如地表地芽孢杆菌或嗜热脂肪地芽孢杆菌)中分离核糖体。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一定量的RNA聚合酶或其片段或变体，RNA聚合酶是一种将DNA序列转录成RNA并存在于各种生物体中的酶。例如，在一个优选实施例中，本发明可包括一种T7 RNA聚合酶，其氨基酸序列为SEQ ID NO.136。T7 RNA聚合酶来源于T7噬菌体，是一种可与称为T7启动子的特异性DNA序列结合，从而将下游DNA序列转录成RNA的酶。除T7 RNA聚合酶之外，还可以在本发明中使用各种RNA聚合酶。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一定量的RNase抑制剂。RNase酶促进将RNA分解成寡核苷酸。RNase抑制剂在本领域众所周知；因此，应包含在重组无细胞表达系统中的RNase抑制剂类型和数量在本领域普通技术人员的技能范围内。RNase抑制剂的非限制性示例包括哺乳动物核糖核酸酶抑制蛋白[如猪核糖核酸酶抑制剂和人核糖核酸酶抑制剂(如人胎盘糖核酸酶抑制剂和重组人核糖核酸酶抑制剂)]、金精三羧酸(ATA)及其盐[如(金精三羧酸三铵盐(铝试剂))]、腺苷5′-焦磷酸、2′-胞苷单磷酸游离酸(2′-CMP)、5′-二磷酸腺苷3′-磷酸(ppA-3′-p)、5′-二磷酸腺苷2′-磷酸(ppA-2′-p)、亮氨酸、聚乙烯磺酸、聚(天冬氨酸)、酪氨酸-谷氨酸聚合物、5′-磷酸-2′-脱氧尿苷3′-焦磷酸P′→5′-酯腺苷3′-磷酸(pdUppAp)及其类似物、衍生物和盐。
在一个实施例中，所述重组无细胞表达系统可包含一种反应混合物，所述反应混合物包含一定量的氨基酸、一个多核苷酸(如通常以质粒合成模板的形式编码一个靶序列的一个mRNA或DNA模板，或线性表达(或合成)模板(LET或LST))以及在“121申请”中确定的其他化合物和序列，所述申请涉及无机多磷酸盐能量再生系统，优选一种偶联AdK/PPK能量再生系统，其可能对积极促进体外表达反应必不可少。
如“121申请”(通过本发明的引用，成为本发明的一部分)中的图8所示，在另一个优选实施例中，可将经分离和纯化的GstAdK(“121申请”(通过本发明的引用，成为本发明的一部分)的SEQ ID NO.8)和/或TaqPPK(“121申请”(通过本发明的引用，成为本发明的一部分)的SEQ ID NO.11)加入此无细胞表达系统(含一定量的无机多磷酸盐)中。在一个实施例中，这种量的无机多磷酸盐可包括最佳多磷酸盐浓度范围。在本优选实施例中，通常将此类最佳多磷酸盐浓度范围定义为保持反应平衡稳定的无机多磷酸盐(PPi)浓度。在本优选实施例中，最佳多磷酸盐浓度范围可约为0.2-2mg/ml PPi。
如上所述，PPK可使用多磷酸盐和AMP合成ADP。在本优选实施例中，Gst AdK和PPK的偶联作用可将两个ADP转化为一个ATP和一个一磷酸腺苷(AMP)，从而去除系统中的腺苷二磷酸(ADP)：
Figure BDA0003398279510000231
这种反应足以快速地促进PPK的平衡反应，产生ADP：
Figure BDA0003398279510000232
在此系统中，存在高浓度AMP可进一步促进TaqPPK反应，产生ADP。
在一个优选实施例中，可在生物反应器系统中使用本发明的重组无细胞系统生产大分子。在本文中，“生物反应器”可以是任何形式的封闭装置，可在体外维持一个有利于产生大分子的环境。生物反应器可在成批、连续或半连续的基础上运行，例如，通过进料反应溶液运行。参照“121申请”(通过本发明的引用，成为本发明的一部分)中的图14，在本实施例中，本发明可进一步包括一种无细胞培养装置。在某些优选实施例中，这种细胞培养装置可培养嗜热菌。在一个优选实施例中，发酵罐可拆卸并且可单独进行高压灭菌。此外，这种无细胞培养装置可实现生物体有氧生长或厌氧生长。此外，无细胞表达生物反应器和无细胞培养装置适用于各种细胞培养，如微藻类、植物细胞等。
在一个实施例中，本发明可特别适用于连续交换式或连续流动式生物反应器(1)操作。在本优选实施例中，这种连续交换式重组生产装置可包括多个中空纤维和中空纤维生物反应器(作为交换介质)，对生物制剂、疫苗、蛋白、酶、生物仿制药进行体外转录、体外翻译和体外生物合成以及在连续流动操作中使用酶对小分子进行生物合成或化学修饰。
参照图5，连续流动式生物反应器装置可包括一个或多个中空纤维(2)和中空纤维生物反应器(2)作为交换介质，对生物制剂、蛋白、酶、生物仿制药进行体外转录、体外翻译和体外生物合成以及在连续流动操作中使用酶对小分子进行生物合成或化学修饰。在本实施例中，可向装置连续供应本文所述的底物，并进一步通过中空纤维反应器(2)内室与外室之间的浓度梯度去除反应产物，还可延长运行时间，并独立于批次生产生物和生物修饰材料，这些材料可从内室的“流通”液中分离。
如图5A和图5B所示，本文描述了示例性中空纤维反应器(2)的操作情况。在本实施例中，当将进料溶液推过反应器内室(3)时，由于纤维具有渗透性，因此可连续供应mRNA合成(核苷酸)底物、通用蛋白(氨基酸)、化合物体外生物合成或化学修饰底物以及本发明从“121申请”中纳入的ATP再生系统(用于提供ATP)和(通过核苷酸激酶(如NDPK))核糖体操作用GTP，而外室(4)则包含各种酶和因子，可促进体外转录、体外翻译以及连续交换中的体外生物合成反应。在图5B所示的闭环系统中分离和纯化所产生的蛋白、酶和大型生物制剂。这种闭环系统可防止和/或降低产物潜在污染、溢出或接触风险，从而减少需加工的体积，并减少任何种类生物制剂的生产空间足迹。直接增加反应容器的体积，可从研究规模的生物合成实现工业规模生产，从而减少开发工作，降低工艺缩放成本，并缩短开发时间。
在本文中，体外重组无细胞表达系指在含生物提取物和/或已定义无细胞反应组分的反应混合物或溶液中无细胞合成多肽。所述反应混合物可包含一个模板或基因模板，用于生产大分子，如DNA和mRNA等；待合成大分子的单体，如氨基酸和核苷酸等；以及进行合成所必需的辅因子、酶及其他试剂，如核糖体、tRNA、聚合酶、转录因子等。可采用批量、连续流动或半连续流动的方式进行所述重组无细胞合成反应和/或添加细胞三磷酸腺苷(ATP)能量再生系统组分(通过本发明的引用，成为本发明的一部分)。
可通过本发明表达的一些靶蛋白可包括但不限于：疫苗、真核肽、原核肽、细菌相关肽、真菌相关肽、酵母相关肽、人类相关肽、植物相关肽、毒素肽、血管活性肠肽、加压素肽、新型或人造工程肽、病毒相关肽、噬菌体相关蛋白、激素、抗体、细胞受体、细胞调节蛋白以及任何上述多肽的片段。
由于本发明涉及生产基因改造生物体并且涉及重组DNA技术，以下定义可协助说明本发明。
在本文中，术语“分离”、“纯化”或“生物纯”系指材料基本上不含在其天然状态或生产时通常包含的组分。在一个示例性实施例中，通常采用分析化学技术确定纯度和均匀性，如聚丙烯酰胺凝胶电泳或高效液相色谱法。基本上对作为制剂优势种的核酸或特定细菌进行纯化。在一个示例性实施例中，术语“纯化”表示核酸或蛋白本质上在电泳凝胶中产生一条谱带。分离核酸或蛋白的纯度通常以一个范围的形式表示。组分的纯度范围下限为约60％、约70％或约80％，纯度范围上限为约70％、约80％、约90％或约90％以上。
在优选实施例中，无细胞表达系统可输出一种产物，如可分离或纯化的肽或其片段等。在实施例中，分离或纯化靶蛋白，其中，通过有机溶剂沉淀(如甲醇、乙醇或丙酮沉淀)、有机盐或无机盐沉淀(如三氯乙酸(TCA)或硫酸铵沉淀)、非离子聚合物沉淀(如聚乙二醇(PEG)沉淀)、pH沉淀、温度沉淀、免疫沉淀、色谱分离(如吸收、离子交换、亲和及凝胶排阻色谱法)、聚焦层析、等电聚焦、高效液相色谱法(HPLC)、凝胶电泳、渗析、微滤等，从反应混合物的至少一种其他组分中至少部分分离所述靶蛋白。
在本文中，术语“活性”系指与全长(完整)蛋白相关的肽或其部分的功能活性或活性。功能活性包括但不限于催化活性或酶活性、抗原性(与多肽结合或与多肽竞争以结合抗多肽抗体的能力)、免疫原性、形成多聚体的能力以及与多肽受体或配体特异性结合的能力。优选地，在0℃-30℃的温度下，所产生蛋白的活性保留至少55％、60％、65％、70％、80％、85％、90％、95％或更多的初始活性至少3天。
在本文中，术语“核酸”系指核糖核苷酸或脱氧核糖核苷酸的聚合物。“核酸”聚合物通常以单链或双链的形式存在，但已知也可形成含两条或多条链的结构。术语“核酸”包括天然核酸聚合物以及含已知核苷酸类似物或修饰骨架残基或键的核酸，此类核酸类似物以合成、天然和非天然形式存在，具有与参考核酸相似的结合特性，且代谢方式与参考核苷酸类似。示例性类似物包括但不限于硫代磷酸酯、氨基磷酸酯、甲基膦酸酯、手性甲基膦酸酯、2-O-甲基核糖核苷酸和肽核酸(PNA)。“DNA”、“RNA”、“多核苷酸”、“多核苷酸序列”、“寡核苷酸”、“核苷酸”、“核酸”、“核酸分子”、“核酸序列”、“核酸片段”和“分离核酸片段”在本文中可互换使用。对于核酸，大小以千碱基(kb)或碱基对(bp)表示。通常通过琼脂糖或丙烯酰胺凝胶电泳、序列核酸或已公布的DNA序列进行估计。对于蛋白，大小以千道尔顿(kDa)或氨基酸残基数量表示。通过凝胶电泳、序列蛋白、衍生氨基酸序列或已公布的蛋白序列估计蛋白大小。
在本文中，术语“靶蛋白”通常系指含5个以上氨基酸的任何肽或蛋白。对于产生细菌细胞的细菌(如宿主菌(如大肠杆菌)中产生的人蛋白或酵母蛋白)，多肽可具有同源性或优选具有外源性，这意味着它们具有异源性，即异质性。优选使用哺乳动物多肽、病毒、细菌、真菌和人造工程多肽。
在本领域中，众所周知，不同的生物体优先利用不同的密码子产生多肽。可在设计编码本发明所述蛋白和嵌合体的核酸分子时利用此类“密码子应用”偏好，以在特定宿主细胞系统中优化表达。
可对本发明所述的所有核苷酸序列进行密码子优化，以在特定生物体中进行表达，或增加产量。密码子优化通常可提高目标基因的翻译效率，从而改善蛋白表达。通过优化自定义设计基因中的密码子应用，也可增加基因功能。在密码子优化实施例中，一个种类中的低频密码子可由高频密码子代替，例如，低频密码子UUA可由亮氨酸的高频密码子CUG代替。密码子优化可增加mRNA稳定性，从而改变蛋白翻译或蛋白折叠速率。此外，密码子优化可自定义转录和翻译控制、修饰核糖体结合位点或稳定mRNA降解位点。
除非另有说明，否则特定核酸序列还隐式包含其适当修饰的变体(例如，简并密码子替换)、互补序列、反向互补序列以及明确指明的序列。具体地说，可通过产生序列，实现简并密码子替换，在此类序列中，一个或多个所选(或所有)密码子的第三位置被混合碱基和/或脱氧肌苷残基替代(见Batzer等人，《核酸研究》，19：5081(1991年)；Ohtsuka等人，《生物化学杂志》，260：2605-2608(1985年)；以及Rossolini等人，《分子细胞探针》，8：91-98(1994))。除核苷酸密码子(编码氨基酸)的简并性质之外，改变多核苷酸会在给定位点产生化学等效的氨基酸，但不会影响已编码多肽的功能特性，这在本领域众所周知。“保守氨基酸替换”是一种预计对参考多肽特性干扰最少的替换。换句话说，保守氨基酸替换基本上保留参考蛋白的结构和功能。因此，氨基酸丙氨酸(一种疏水性氨基酸)的密码子可由编码另一种低疏水性残基(如甘氨酸)或高疏水性残基(如缬氨酸、亮氨酸或异亮氨酸)的密码子替代。同样，改变会导致一个负电荷残基替代另一个负电荷残基，如天冬氨酸替代谷氨酸，或一个正电荷残基替代另一个正电荷残基，如赖氨酸替代精氨酸或组氨酸，此类改变预计也可产生功能等效蛋白或多肽。本领域的一般技术人员均了解示例性保守氨基酸替换事宜。保守氨基酸替换通常保持(a)替换区域多肽骨架的结构，例如，β折叠或α螺旋构象；(b)替换位点分子的电荷或疏水性；和/或(c)大量侧链。
可使用任何同源性比较软件(计算双序列比对)确定同源性(例如，同源性百分比，序列一致性+序列相似性)。在本文中，在两个核酸或多肽序列的情况下，“序列一致性”或“一致性”包括引用两个序列(在比对时相同)中的残基。如果在引用蛋白时使用序列一致性百分比，则应认识到，不同残基位置的保守氨基酸替换通常也不同，其中，氨基酸残基被具有相似化学特性(例如电荷或疏水性)的其他氨基酸残基替代，因此不会改变分子的功能特性。如果序列的保守替换不同，则序列一致性可向上调整，以校正替换的保守性质。此类保守替换不同的各个序列应具有“序列相似性”或“相似性”。本领域的技术人员均熟知进行这种调整的方式。其中通常涉及将保守替换评为一部分不匹配，而非完全不匹配，从而提高序列一致性百分比。因此，如果相同氨基酸的得分为1，非保守替换的得分为0，则保守替换的得分在0-1之间。例如，根据Henikoff S和Henikoff JG算法计算保守替换的得分。[蛋白块中的氨基酸替换矩阵。《美国国家科学院院刊》1992期，89(22)：10915-9]。
根据一个特定实施例，同系物序列至少与本文提供的序列(核酸或氨基酸序列)60％、65％、70％、75％、80％、85％、90％、95％、99％相同或甚至完全相同。50％-99％的SEQID NO.1-22同系物序列可包括在本发明的某些实施例中。
在本文中，术语“引物”系指在适当条件下能够作为DNA合成起始点的寡核苷酸。此类条件包括可实现以下目的的条件：在适当温度下，在适当缓冲液中存在四种不同的核苷三磷酸和一种延伸剂(例如，一种DNA聚合酶或逆转录酶)时，诱导合成与核酸链互补的引物延伸产物。
引物优选单链DNA。引物的适当长度取决于引物的预期用途，但通常范围为约6个到约225个核苷酸，包括中等范围，如15个到35个核苷酸、18到75个核苷酸和25个到150个核苷酸。短引物分子通常需要更低的温度，才能使用模板形成充分稳定的杂化复合物。引物不需要反映核酸模板的准确序列，但必须充分互补，以使用模板进行杂化。给定靶序列扩增用适当引物的设计在本领域众所周知，在本文所引用的文献中对此进行了说明。
在本文中，“聚合酶”系指催化核苷酸聚合作用的一种酶。“DNA聚合酶”催化脱氧核糖核苷酸的聚合作用。已知的DNA聚合酶包括强烈火球菌(Pfu)DNA聚合酶、大肠杆菌DNA聚合酶I、T7 DNA聚合酶和水生栖热菌(Taq)DNA聚合酶等。“RNA聚合酶”催化核糖核苷酸的聚合作用。DNA聚合酶的上述示例也称为DNA依赖性DNA聚合酶。RNA依赖性DNA聚合酶也在DNA聚合酶的范围内。逆转录酶(包括由逆转录病毒编码的病毒聚合酶)是RNA依赖性DNA聚合酶的一个示例。RNA聚合酶(“RNAP”)的已知示例包括T3 RNA聚合酶、T7 RNA聚合酶、SP6 RNA聚合酶和大肠杆菌RNA聚合酶等。RNA聚合酶的上述示例也称为DNA依赖性RNA聚合酶。可通过本领域众所周知的方式确定上述任何一种酶的聚合酶活性。
在本文中，术语“反应混合物”、“无细胞反应混合物”或“重组无细胞反应混合物”系指含有执行给定反应所需试剂的一种溶液。无细胞表达系统“反应混合物”或“反应溶液”通常含有粗提取物或部分纯化提取物(如细菌、植物细胞、微藻类、真菌或哺乳动物细胞的提取物)、核苷酸翻译模板以及一种从翻译模板中促进无细胞蛋白合成的适当反应缓冲液。一方面，CF反应混合物可包含一个外源性RNA翻译模板。在其他方面，CF反应混合物可包含一个编码开放读码框架(可连接DNA依赖性RNA聚合酶的启动子元件)的DNA表达模板。在这些其他方面，CF反应混合物也可包含一种DNA依赖性RNA聚合酶，直接转录编码开放读码框架的RNA翻译模板。在这些其他方面，CF反应混合物可包含其他NTP和二价阳离子辅因子。如果反应混合物含有执行反应所需的所有试剂，则将其称为完全反应混合物，如果仅含有一小部分的必要试剂，则将其称为不完全反应混合物。本领域的普通技术人员理解，为方便起见，同时也为了确保储存稳定性，或根据具体应用对组分浓度进行调整，反应组分通常以单独溶液的形式储存，每种溶液分别包含总组分的一小部分，将在反应前组合反应组分，以形成完全反应混合物。此外，本领域的普通技术人员也理解，为了实现商业化，将单独包装反应组分，有用的商用试剂盒可包含本发明反应组分的任何一小部分。此外，这些普通技术人员理解，反应混合物中的一些组分在用于某些实施例时，不一定会产生无细胞表达产物。术语“无细胞表达产物”可以是通过无细胞表达系统生产的任何生物制品。
术语“约”或“大约”系指在一个值或多个值具有统计意义的范围内，如规定浓度、长度、分子量、pH值、时间范围、温度、压力或体积。此类值或范围可在一个数量级内，通常在给定值或范围的20％内，更通常在10％内，甚至更通常在5％内。“约”或“大约”所涵盖的允许偏差取决于所研究的特定系统。除非另有说明，否则，术语“包括(comprising)”、“具有(having)”和“包含(including)”应视为开放式术语(即，系指“包括但不限于”)。
除非本文另有说明，否则，本文中对值范围的复述仅用作单独引用范围内每个单独值的一种速记法，并且包括定义此范围的终点界限，每个单独值均被纳入本说明书中，如同在本文中单独列举一样。
当涉及细胞或核酸、蛋白或载体时，术语“重组”或“基因修饰”系指通过引入异源核酸或蛋白或改变天然核酸或蛋白，对细胞、生物体、核酸、蛋白或载体进行修饰，或细胞由经此类修饰的细胞衍生而来。因此，重组细胞可表达未在原生(非重组或野生型)形式的细胞内发现的基因，或表达异常表达、过表达、低表达或完全未表达的原生基因。
在本文中，术语“转化”或“基因修饰”系指将一个或多个核酸分子转移到细胞中。当稳定复制核酸分子时，通过转导到细菌、细胞或生物体的核酸分子，对微生物进行“转化”或“基因修饰”。在本文中，术语“转化”或“基因修饰”包含可将核酸分子引入细胞或生物体(如细菌)中的所有技术。
在本文中，术语“启动子”系指DNA的一个区域，此区域可能是开始转录的上游，也可能参与RNA聚合酶及其他蛋白的识别和结合，以启动转录。启动子可连接编码序列(在细胞中表达)，或启动子可连接编码信号序列的核苷酸序列，此类信号序列可连接编码序列(在细胞中表达)。
当涉及调控序列和编码序列时，术语“可连接”系指调控序列影响所连接的编码序列的表达。“调控序列”或“控制元件”系指影响转录的时间以及水平和数量、RNA加工或稳定性或相关编码序列翻译的核苷酸序列。调控序列可包括启动子；翻译增强序列；内含子；增强子；茎-环结构；阻遏子或结合序列；终止序列；聚腺苷酸化识别序列等。特定调控序列可位于其所连编码序列的上游和/或下游。此外，可连接编码序列的特定调控序列可位于双链核酸分子的相关互补链上。
在本文中，术语“基因组”系指在细胞核内发现的染色体DNA，也系指在细胞的亚细胞组分内发现的细胞器DNA。应用于细菌时，术语“基因组”系指细菌细胞内的染色体和质粒。在本发明的一些实施例中，可将DNA分子引入细菌中，从而将DNA分子整合到细菌的基因组中。在这些及其他实施例中，DNA分子可进行染色体整合，或定位为稳定质粒或位于稳定质粒中。
术语“基因”或“序列”系指可连接适当调控序列(能够以某种方式调控基因产物(如多肽或功能性RNA)的表达)的编码区。基因在编码区(开放读码框架(ORF))前(上游)后(下游)包含DNA的未翻译调控区(如启动子、增强子、阻遏子)，如适用，还在单独编码区(即外显子)之间包含介入序列(即内显子)。在本文中，术语“结构基因”系指转录成mRNA，然后翻译成氨基酸序列(具有特异性多肽特性)的一种DNA序列。
在本文中，术语“表达”或“编码序列表达”(例如，基因或转基因)系指一个过程，通过这一过程，将核酸转录单位(包括基因组DNA或cDNA)的已编码信息转化成细胞的可操作、不可操作或结构部分。基因表达可能受到外部信号的影响；例如，细胞、组织或生物体接触一种可增加或减少基因表达的药剂。也可在DNA-RNA-蛋白通路中的任何位置调控基因表达。例如，通过控制转录、翻译、RNA运输和加工、中间分子(如mRNA)的降解或在蛋白分子产生后通过蛋白分子的活化、失活、区室化或降解或其组合，实现基因表达调控。可采用本领域的任何已知方法，在RNA水平或蛋白水平上测量基因表达，包括但不限于诺瑟杂交法、RT-PCR、蛋白免疫印迹法或体外、在体或体内蛋白活性检测。
术语“载体”系指将DNA、RNA、蛋白或多肽引入宿主中的某种手段。引入宿主中的多核苷酸、蛋白和多肽可具有治疗性或预防性；可编码或可以是一种抗原；本质上可调控等。存在各种类型的载体，包括病毒、质粒、噬菌体、粘粒和细菌。
“表达载体”是一种能够在所选宿主细胞或生物体中进行复制的核酸。表达载体可作为自主结构进行复制，或可全部或部分整合到宿主细胞染色体或细胞器核酸中，或用作向细胞输送异质DNA的穿梭载体，从而与宿主细胞基因组一起复制。因此，表达载体是一种多核苷酸，能够在所选宿主细胞、细胞器或生物体中进行复制，如质粒、病毒、人工染色体、核酸片段，表达载体上的某些基因(包括目标基因)在细胞、细胞器或生物体内转录并翻译成多肽或蛋白；或是本领域已知的任何适当结构，其中包含“表达盒”。相比之下，如本文示例所述，“盒”是一种多核苷酸，含有本发明所述表达载体的一部分。使用此盒有助于组装表达载体。表达载体是一种复制子，如质粒、噬菌体、病毒、嵌合病毒或粘粒，包含可连接表达控制序列的所需多核苷酸序列。
术语“表达产物”涉及在本文所述无细胞表达系统中表达的蛋白，因此可与含5个以上氨基酸的任何肽或蛋白互换使用，并通常系指此类肽或蛋白。对于从中衍生无细胞提取物的生物体(如无细胞提取物中产生的人蛋白、植物蛋白、病毒蛋白、酵母蛋白等)，多肽可具有同源性或外源性，这意味着它们具有异源性，即异质性。在一些实施例中，术语“衍生”系指从细菌中提取或表达和分离。例如，在一个实施例中，蛋白可由嗜热菌衍生而来，这可能意味着蛋白对嗜热菌具有内源性，并从所述细菌中分离，或在不同细菌中进行异源性表达，并分离为单独的蛋白或细胞提取物。
“无细胞提取物”或“裂解物”可由各种生物体和或细胞衍生而来，包括细菌、嗜热菌、耐热菌、古菌、厚壁菌门、真菌、藻类、微藻类、植物细胞培养物和植物悬浮培养物。
在本文中，除非上下文另有明确说明，否则单数形式“一个(a)”、“一个(an)”和“所述(the)”也包括复数指称对象。因此，对“细胞”的引用包括多个此类细胞，对“培养物”的引用包括对本领域技术人员已知的一种或多个培养物及其等同物的引用。除非另有明确说明，否则本文所用的所有技术和科学术语均具有本发明所属领域普通技术人员公知的相同含义。
通过引用以下示例，更容易理解本发明，这些示例仅旨在说明本发明所述实施例的某些方面。这些示例不旨在限制本发明，因为本领域的技术人员将从上述教导和以下示例中认识到，其他技术和方法也可满足权利要求，在不脱离本发明范围的情况下，均可使用。的确，尽管已经参照本发明的优选实施例具体显示和描述了本发明，但是本领域的技术人员应理解，在不脱离所附权利要求所涵盖的本发明范围的情况下，可以对其形式和细节进行各种修改。
示例
示例1：重组无细胞表达系统的蛋白合成和克隆。
发明人优选从所选嗜热菌中合成多个核心重组蛋白，并将它们克隆到所选表达载体中，用于在重组无细胞表达系统中使用。在本实施例中，发明人合成多个核心重组嗜热起始因子(IF)，并将它们克隆到所选表达载体中。在本实施例中，发明人合成多个核心重组嗜热延伸因子(EF)，并将它们克隆到所选表达载体中。在本实施例中，发明人合成多个核心重组释放因子(RF)，并将它们克隆到所选表达载体中。在本实施例中，发明人合成至少一个核心重组核糖体循环因子(RRF)，并将它们克隆到所选表达载体中。在本实施例中，发明人合成多个核心重组氨酰-tRNA合成酶(RS)，并将它们克隆到所选表达载体中。在本实施例中，发明人合成至少一个核心重组甲硫氨酰-tRNA转甲酰基酶(MTF)，并将它们克隆到所选表达载体中。
如表1所示，在一个优选实施例中，发明人合成、克隆、在大肠杆菌中表达并纯化至少十二(12)个不同的重组因子(包括核苷酸和/或氨基酸序列)以及至少二十二(22)个重组合成酶(包括核苷酸和/或氨基酸序列(SEQ ID NO.1-132))，形成示例性核心重组蛋白混合物，其中包含至少三十四(34)个可应用于创新性重组无细胞表达系统的蛋白。发明人将这些核心蛋白克隆到表达载体中，例如，pET151/D-TOPO(pET151)、pET24a(+)或pNAT(如图7-8和图9所示)。
发明人进一步获得一种重组无细胞反应混合物，其中包含三十四(34)个已确定蛋白中的一个或多个蛋白以及来自示例性嗜热菌的所选分离核糖体和tRNA。接下来，发明人向重组无细胞反应混合物中添加了一定量的RNA聚合酶(特别是T7 RNA聚合酶)以及示例性氨基酸和缓冲液。如上所述，发明人进一步获得一种重组无细胞反应混合物，其中包含在第PCT/US201 8/012121号PCT申请(“121申请”)中确定的无机多磷酸盐能量再生系统的一种或多种组分。
示例2：产生示例性重组无细胞反应混合物。
在一个实施例中，发明人获得一种能够体外转录和翻译的重组无细胞反应混合物，从包括以下几项的一组中选择所述重组无细胞反应混合物：
-.包含表1中确定的至少三十三(33)个嗜热核心蛋白的一种反应混合物；
-表1中确定的大肠杆菌的一(1)个蛋白；
-来自嗜热菌的tRNA；
-从所选嗜热菌中分离的一定量核糖体；
-.一定量的氨基酸；
-一定量的核苷三磷酸(NTP)，如ATP、CTP、GTP、TTP；
-一定量的反应缓冲液；以及
-在“121申请”(已纳入本发明中)的权利要求、附图、序列和说明书中确定的无机多磷酸盐能量再生系统或能量再生系统的一种或多种组分。
示例3：重组氨酰-tRNA合成酶的活性。
发明人确认了每个纯化氨酰-tRNA合成酶(RS)的活性。氨酰-tRNA合成酶反应过程通常分为两个步骤：
步骤1：活化 氨基酸+ATP＝＞氨酰-AMP+PPi
步骤2：转移 氨酰-AMP+tRNA＝＞氨酰-tRNA+AMP
可使用EnzCheck焦磷酸盐试剂盒对获得的PPi进行测量。利用这一大纲，发明人使用商用焦磷酸盐检测试剂盒(EnzCheck焦磷酸盐检测试剂盒、分子探针、E-6654(通过本发明的引用，成为本发明的一部分))进行了动力学检测。这种商用检测可采用分光光度法间接测量焦磷酸盐的酶法生产情况。在30μl的总量下设置每个RS反应，最终浓度如表2所示。如表3所示，使用12.5μl的RS反应混合物设置焦磷酸盐检测的50μl反应。在96孔板中设置焦磷酸盐检测，并以2分钟的间隔在读板器(设置用于读取360nm处的吸光度)上自动读取。这些动力学测量用作所有RS蛋白活性和功能的定性首要检测。
根据制造商说明书进行检测，绘制每个RS的吸光度随时间变化的图表。如图1和图2所示，每个RS均显示了良好的活性(无tRNA作为对照)，由于ATP水解成ADP+Pi，产生了无机焦磷酸盐，使用EnzCheck检测试剂盒可间接检测Pi。即使在低吸光度变化的情况下，图1和图2中的数据也与有关RS的已发表报告以及有关ATP使用的其他酶动力学的所示图表(由生产指南提供)类似。为清晰起见，对于图1和图2，仅在每个图上绘制10个RS，但源于同样的实验。
可使用AMP-GloTM试剂盒测量从氨酰-tRNA合成酶反应中获得的AMP。发明人使用商用AMP检测试剂盒(AMP-GloTM检测试剂盒、Promega V5012(通过本发明的引用，成为本发明的一部分))进行检测。这种商用检测通过发光反应间接测量AMP的酶法生产情况。可使用已纳入的一项标准来校准和计算AMP的产量。这种检测是一种定量终点测量检测。在100μl的总量下设置每个RS反应，最终浓度如图4所示，并在37℃下运行一小时。然后，根据制造商说明书重复进行AMP检测，并使用标准曲线计算产生的AMP(图17B)。图17A显示了利用示例性tRNA(来自大肠杆菌)的三种独立氨酰-tRNA合成酶AMP生产活性检测的结果。标准AMP曲线如图17B所示。
示例4：确认重组氨酰-tRNA合成酶的活性。
发明人还对每个克隆RS的活性进行了额外确认，即，使用商用试剂盒(Cayman，孔雀石绿磷酸盐检测试剂盒，#10009325(通过本发明的引用，成为本发明的一部分))进行孔雀绿磷酸盐检测。所产生的焦磷酸盐将与孔雀石绿形成复合物，导致颜色发生变化，可通过吸光度来衡量这一点。可使用已纳入的一项标准来校准和计算PPi的产量。这种检测是一种定量终点测量检测。根据制造商说明书进行所有反应，并使用标准曲线(在图上显示为小入口)计算所产生的PPi。
如下表4所示，每个RS反应的最终浓度包括总体积150μl。在此检测中，使用来自大肠杆菌的示例性tRNA。图3A表明，与不含反应缓冲液(无ATP)以及一个RS(AsnRS+Arg)的错误氨基酸的对照品相比，所有RS均具有良好活性。以相同的摩尔浓度使用每个RS，并在使用试剂盒测量PPi浓度前，培养60分钟。对每个柱进行本底/空白测量校正，这些柱表示重复测量的平均值。如图3B所示，根据上文中的说明，使用来自芽孢杆菌嗜热菌(如地表地芽孢杆菌或嗜热脂肪地芽孢杆菌)的tRNA重复进行相同检测。
示例5：示例性蛋白的重组无细胞表达。
发明人演示了在本发明的重组无细胞表达系统中生产两个示例性GFP肽(SEQ IDNO.134-135)的情况。如表6所示，产生对照和模板重组无细胞表达混合物。图11-14显示了分离以下表6中确定的核心重组蛋白的情况。如图4所示，重组无细胞表达系统对所添加的模板DNA进行转录，并将获得的mRNA翻译成蛋白(如图4中的条带所示)。如图15进一步所示，发明人演示了利用本文所述的重组无细胞表达系统实时生产荧光蛋白(muGFP；SEQ IDNO.134)产物的情况。如图16进一步所示，发明人演示了利用本文所述的重组无细胞表达系统生产荧光蛋白(deGFP，SEQ ID NO.135)产物的情况。此外，发明人演示了通过逆纯化从所产生的GFP肽中去除重组无细胞表达系统翻译组分的情况。如图16具体所示，在逆纯化后，使用无细胞蛋白表达反应的抗FLAG抗体执行蛋白免疫印迹法。
表格
表1：重组无细胞表达系统的示例性核心蛋白
Figure BDA0003398279510000341
表2：焦磷酸盐检测RS反应混合物浓度。
反应缓冲液 RS反应混合物(30μl)
50mM HEPES 1mM ATP
150mM NaCl 20μg tRNA
10mM KCl 2mM氨基酸
5mM MgSO4 7μg RS
2mM DTT 1x反应缓冲液
ddH2O
表3：50μl焦磷酸盐检测反应。
焦磷酸盐检测(50μl)
1x反应缓冲液
0.4mMMESG底物
1U嘌呤核苷磷酸化酶
0.03U无机焦磷酸盐酶
12.5μl RS反应混合物
ddH2O
表4：AMP检测RS反应混合物浓度
反应缓冲液 RS反应混合物(100μl)
50mM HEPES 50μM ATP
150mM NaCl 100μg tRNA
10mM KCl 1mM氨基酸
5mM MgSO4 5μg RS
2mM DTT 1x反应缓冲液
ddH2O
表5：重组无细胞蛋白表达反应混合物
Figure BDA0003398279510000351
表6：蛋白、载体和标签组合列表
Figure BDA0003398279510000352
Figure BDA0003398279510000361
Figure BDA0003398279510000371
表7：与地表地芽孢杆菌91A1菌株序列的序列一致性
Figure BDA0003398279510000372
Figure BDA0003398279510000381
参考文献
以下参考文献通过本发明的整体引用，成为本发明的一部分：
[1]Carlson，Erik D.等人。“无细胞蛋白合成：应用的时代到来了”。《生物技术进展》，30.5(2012)：1185-1194。PMC.Web.2018年1月1日。
[2]Lloyd，A.J.，Thomann，H.U.，Ibba，M.和Soil，D.(1995年)。用于测量氨酰-tRNA合成酶活性的广泛适用连续分光光度检测法。《核酸研究》，23(15)，2886-2892。
序列列表
SEQ ID NO.1
DNA
IF-1-GbIF-1-EcOpt
芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000391
SEQ ID NO.2
氨基酸
IF-1-GbIF-1-EcOpt
芽孢杆菌
MAKDDVIEVEGTVIETLPNAMFRVELENGHTVLAHVSGKIRMHFIRILPGDRVTVELSPYDLTRGRITYRYK
SEQ ID NO.3
DNA
IF-2-GsIF-2-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000392
Figure BDA0003398279510000401
SEQ ID NO.4
氨基酸
IF-2-GsIF-2-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000411
SEQ ID NO.5
DNA
IF-3-GbIF-3-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000412
SEQ ID NO.6
氨基酸
IF-3-GbIF-3-EcOpt
地芽孢杆菌
Figure BDA0003398279510000421
SEQ ID NO.7
DNA
EF-G-GsEF-G-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000422
Figure BDA0003398279510000431
SEQ ID NO.8
氨基酸
EF-G-GsEF-G-EcOpt
地芽孢杆菌
Figure BDA0003398279510000432
Figure BDA0003398279510000441
SEQ ID NO.9
DNA
EF-Tu-GsEF-Tu-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000442
SEQ ID NO.10
氨基酸
EF-Tu-GsEF-Tu-EcOpt
地芽孢杆菌
Figure BDA0003398279510000451
SEQ ID NO.11
DNA
EF-Ts-GsEF-Ts-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000452
SEQ ID NO.12
氨基酸
EF-Ts-GsEF-Ts-EcOpt
地芽孢杆菌
Figure BDA0003398279510000461
SEQ ID NO.13
DNA
EF-4-GsEF-4-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000462
Figure BDA0003398279510000471
SEQ ID NO.14
氨基酸
EF-4-GsEF-4-EcOpt
地芽孢杆菌
Figure BDA0003398279510000472
Figure BDA0003398279510000481
SEQ ID NO.15
DNA
EF-P-GsEF-P-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000482
SEQ ID NO.16
氨基酸
EF-P-GsEF-P-EcOpt
地芽孢杆菌
Figure BDA0003398279510000483
ADGTYVSRA
SEQ ID NO.17
DNA RF-1
标题： GsRF-1-EcOpt
来源： 嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000484
Figure BDA0003398279510000491
SEQ ID NO.18
氨基酸
RF-1-GsRF-1-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000492
SEQ ID NO.19
DNA
RF-2-GsRF-2-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000501
SEQ ID NO.20
氨基酸
RF-2-GsRF-2-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000502
Figure BDA0003398279510000511
SEQ ID NO.21
DNA
RF-3-BX1-RF-3-EcOpt
芽孢杆菌XI(大肠杆菌密码子优化)
Figure BDA0003398279510000512
Figure BDA0003398279510000521
SEQ ID NO.22
氨基酸
RF-3
BX1-RF-3-EcOpt
芽孢杆菌XI
Figure BDA0003398279510000522
SEQ ID NO.23
DNA
RRF-GbRRF-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000523
Figure BDA0003398279510000531
SEQ ID NO.24
氨基酸
RRF
GbRRF-EcOpt
地芽孢杆菌
Figure BDA0003398279510000532
SEQ ID NO.25
DNA
AlaRS-GsAlaRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000533
Figure BDA0003398279510000541
Figure BDA0003398279510000551
SEQ ID NO.26
氨基酸
AlaRS-GsAlaRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000552
SEQ ID NO.27
DNA
ArgRS-GsArgRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000553
Figure BDA0003398279510000561
SEQ ID NO.28
氨基酸
ArgRS-GsArgRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000571
SEQ ID NO.29
DNA
AsnRS-GsAsnRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000572
Figure BDA0003398279510000581
SEQ ID NO.30
氨基酸
AsnRS-GsAsnRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000582
SEQ ID NO.31
DNA
AspRS-GsAspRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000583
Figure BDA0003398279510000591
SEQ ID NO.32
氨基酸
AspRS-GsAspRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000601
SEQ ID NO.33
DNA
CysRS-GsCysRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000602
Figure BDA0003398279510000611
SEQ ID NO.34
氨基酸
CysRS-GsCysRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000612
SEQ ID NO.35
DNA
GlnRS-EcGlnRS-EcOpt
大肠杆菌
Figure BDA0003398279510000613
Figure BDA0003398279510000621
SEQ ID NO.36
氨基酸
GlnRS-EcGlnRS-EcOpt
大肠杆菌
Figure BDA0003398279510000631
SEQ ID NO.37
DNA
GluRS-GsGluRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000632
Figure BDA0003398279510000641
SEQ ID NO.38
氨基酸
GluRS-GsGluRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000642
SEQ ID NO.39
DNA
GlyRS-GsGlyRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000643
Figure BDA0003398279510000651
SEQ ID NO.40
氨基酸
GlyRS-GsGlyRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000652
Figure BDA0003398279510000661
SEQ ID NO.41
DNA
HisRS-GsHisRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000662
Figure BDA0003398279510000671
SEQ ID NO.42
氨基酸
HisRS-GsHisRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000672
SEQ ID NO.43
DNA
IIeRS-GsIIeRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000673
Figure BDA0003398279510000681
Figure BDA0003398279510000691
SEQ ID NO.44
氨基酸
IIeRS-GsIIeRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000692
SEQ ID NO.45
DNA
LeuRS-GsLeuRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000701
Figure BDA0003398279510000711
SEQ ID NO.46
氨基酸
LeuRS-GsLeuRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000712
Figure BDA0003398279510000721
SEQ ID NO.47
DNA
IysRS-GsIysRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000722
Figure BDA0003398279510000731
SEQ ID NO.48
氨基酸
IysRS-GsIysRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000732
SEQ ID NO.49
DNA
MetRS-GsMetRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000733
Figure BDA0003398279510000741
SEQ ID NO.50
氨基酸
MetRS-GsMetRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000742
Figure BDA0003398279510000751
SEQ ID NO.51
DNA
Phe-aRS-GsPhe-aRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000752
Figure BDA0003398279510000761
SEQ ID NO.52
氨基酸
Phe-aRS-GsPhe-aRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000762
SEQ ID NO.53
DNA
Phe-bRS-GsPhe-bRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000763
Figure BDA0003398279510000771
Figure BDA0003398279510000781
SEQ ID NO.54
氨基酸
Phe-bRS-GsPhe-bRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000782
SEQ ID NO.55
DNA
ProRS-GsProRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000783
Figure BDA0003398279510000791
SEQ ID NO.56
氨基酸
ProRS-GsProRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000792
Figure BDA0003398279510000801
SEQ ID NO.57
DNA
SerRS-GsSerRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000802
Figure BDA0003398279510000811
SEQ ID NO.58
氨基酸
SerRS-GsSerRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000812
SEQ ID NO.59
DNA
ThrRS-GsThrRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000813
Figure BDA0003398279510000821
SEQ ID NO.60
氨基酸
ThrRS-GsThrRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000831
SEQ ID NO.61
DNA
TrpRS-GsTrpRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000832
Figure BDA0003398279510000841
SEQ ID NO.62
氨基酸
TrpRS-GsTrpRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000842
SEQ ID NO.63
DNA
TyrRS-GsTyrRS-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000843
Figure BDA0003398279510000851
SEQ ID NO.64
氨基酸
TyrRS-GsTyrRS-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000852
SEQ ID NO.65
DNA
ValRS-GsVaIRS-EcOpt
地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000853
Figure BDA0003398279510000861
Figure BDA0003398279510000871
SEQ ID NO.66
氨基酸
ValRS-GsValRS-EcOpt
地芽孢杆菌
Figure BDA0003398279510000872
Figure BDA0003398279510000881
SEQ ID NO.67
DNA
MTF-GsMTF-EcOpt
嗜热脂肪地芽孢杆菌(大肠杆菌密码子优化)
Figure BDA0003398279510000882
SEQ ID NO.68
氨基酸
MTF-GsMTF-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510000883
Figure BDA0003398279510000891
SEQ ID NO.69
DNA
IF-1-GsuIF-1
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000892
SEQ ID NO.70
氨基酸
IF-1-GsuIF-1
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000893
SEQ ID NO.71
DNA
IF-2-GsuIF-2
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000894
Figure BDA0003398279510000901
Figure BDA0003398279510000911
SEQ ID NO.72
氨基酸
IF-2-GsuIF-2
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000912
SEQ ID NO.73
DNA
IF-3-GsuIF-3
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000913
Figure BDA0003398279510000921
SEQ ID NO.74
氨基酸
IF-3-GsuIF-3
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000922
SEQ ID NO.75
DNA
EF-G-GsuEF-G
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000923
Figure BDA0003398279510000931
SEQ ID NO.76
氨基酸
EF-G-GsuEF-G
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000932
Figure BDA0003398279510000941
SEQ ID NO.77
DNA
EF-Tu-GsuEF-Tu
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000942
Figure BDA0003398279510000951
SEQ ID NO.78
氨基酸
EF-Tu-GsuEF-Tu
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000952
SEQ ID NO.79
DNA
EF-Ts-GsuEF-Ts
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000953
Figure BDA0003398279510000961
SEQ ID NO.80
氨基酸
EF-Ts-GsuEF-Ts
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000962
SEQ ID NO.81
DNA
EF-4-GsuEF-4
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000963
Figure BDA0003398279510000971
SEQ ID NO.82
氨基酸
EF-4-GsuEF-4
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000972
Figure BDA0003398279510000981
SEQ ID NO.83
DNA
EF-P-GsuEF-P
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000982
SEQ ID NO.84
氨基酸
EF-P-GsuEF-P
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000983
SEQ ID NO.85
DNA
RF-1-GsuRF-1
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000991
SEQ ID NO.86
氨基酸
RF-1-GsuRF-1
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510000992
Figure BDA0003398279510001001
SEQ ID NO.87
DNA
RF-2-Gsu-RF2
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001002
SEQ ID NO.88
氨基酸
RF-2-GsuRF-2
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001003
Figure BDA0003398279510001011
SEQ ID NO.89
DNA
RRF-GsuRRF
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001012
SEQ ID NO.90
氨基酸
RRF-GsuRRF
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001013
SEQ ID NO.91
DNA
AlaRS-GsuAlaRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001021
Figure BDA0003398279510001031
SEQ ID NO.92
氨基酸
AlaRS-GsuAlaRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001032
Figure BDA0003398279510001041
SEQ ID NO.93
DNA
ArgRS-GsuArgRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001042
Figure BDA0003398279510001051
SEQ ID NO.94
氨基酸
ArgRS-GsuArgRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001052
SEQ ID NO.95
DNA
AsnRS-GsuAsnRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001061
SEQ ID NO.96
氨基酸
AsnRS-GsuAsnRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001062
Figure BDA0003398279510001071
SEQ ID NO.97
DNA
AspRS-GsuAspRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001072
Figure BDA0003398279510001081
SEQ ID NO.98
氨基酸
AspRS-GsuAspRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001082
SEQ ID NO.99
DNA
CysRS-GsuCysRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001083
Figure BDA0003398279510001091
SEQ ID NO.100
氨基酸
CysRS-GsuCysRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001092
Figure BDA0003398279510001101
SEQ ID NO.101
DNA
GluRS-GsuGluRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001102
Figure BDA0003398279510001111
SEQ ID NO.102
氨基酸
GluRS-GsuGluRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001112
SEQ ID NO.103
DNA
GlyRS-GsuGlyRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001113
Figure BDA0003398279510001121
SEQ ID NO.104
氨基酸
GlyRS-GsuGlyRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001122
SEQ ID NO.105
DNA
HisRS-GsuHisRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001131
SEQ ID NO.106
氨基酸
HisRS-GsuHisRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001132
Figure BDA0003398279510001141
SEQ ID NO.107
DNA
IleRS-GsuIleRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001142
Figure BDA0003398279510001151
SEQ ID NO.108
氨基酸
IIeRS-GsuIIeRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001161
SEQ ID NO.109
DNA
LeuRS-GsuLeuRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001162
Figure BDA0003398279510001171
Figure BDA0003398279510001181
SEQ ID NO.110
氨基酸
LeuRS-GsuLeuRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001182
SEQ ID NO.111
DNA
LysRS-GsuLysRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001191
SEQ ID NO.112
氨基酸
LysRS-GsuLysRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001201
SEQ ID NO.113
DNA
MetRS-GsuMetRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001202
Figure BDA0003398279510001211
SEQ ID NO.114
氨基酸
MetRS-GsuMetRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001212
Figure BDA0003398279510001221
SEQ ID NO.115
DNA
Phe-aRS-GsuPhe-aRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001222
SEQ ID NO.116
氨基酸
Phe-aRS-GsuPhe-aRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001231
SEQ ID NO.117
DNA
Phe-bRS-GsuPhe-bRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001232
Figure BDA0003398279510001241
SEQ ID NO.118
氨基酸
Phe-bRS-GsuPhe-bRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001251
SEQ ID NO.119
DNA
ProRS-GsuPrORS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001252
Figure BDA0003398279510001261
SEQ ID NO.120
氨基酸
PrORS-GsuPrORSt
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001262
Figure BDA0003398279510001271
SEQ ID NO.121
DNA
SerRS-GsuSerRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001272
Figure BDA0003398279510001281
SEQ ID NO.122
氨基酸
SerRS-GsuSerRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001282
SEQ ID NO.123
DNA
ThrRS-GsuThrRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001283
Figure BDA0003398279510001291
SEQ ID NO.124
氨基酸
ThrRS-GsuThrRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001301
SEQ ID NO.125
DNA
TrpRS-GsuTrrpRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001302
Figure BDA0003398279510001311
SEQ ID NO.126
氨基酸
TrpRS-GsTrpRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001312
SEQ ID NO.127
DNA
TyrRS-GsuTyrRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001313
Figure BDA0003398279510001321
SEQ ID NO.128
氨基酸
TyrRS-GsuTyrRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001322
SEQ ID NO.129
DNA
ValRS-GsuValRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001323
Figure BDA0003398279510001331
Figure BDA0003398279510001341
SEQ ID NO.130
氨基酸
ValRS-GsuValRS
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001342
Figure BDA0003398279510001351
SEQ ID NO.131
DNA
MTF-GsuMTF
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001352
SEQ ID NO.132
氨基酸
MTF-GsuMTF
地表地芽孢杆菌DSM 13552(91A1)
Figure BDA0003398279510001361
SEQ ID NO.133
氨基酸
RF-1-Mut-GsRF-1-EcOpt
嗜热脂肪地芽孢杆菌
Figure BDA0003398279510001362
SEQ ID NO.134
氨基酸
muGFP+His6标签+C标签
维多利亚多管发光水母
Figure BDA0003398279510001363
SEQ ID NO.135
氨基酸
deGFP
维多利亚多管发光水母
Figure BDA0003398279510001371
SEQ ID NO.136
氨基酸
T7 RNA聚合酶
T7噬菌体
Figure BDA0003398279510001372
序列表
<110> 自然工具箱公司(NATURE'S TOOLBOX, INC)
迈克尔·亨伯特(HUMBERT, Michael)
亚历山大·科格林(KOGLIN, Alexander)
查理·维拉纽瓦(VILLANUEVA,Charlie)
<120> 利用嗜热蛋白的重组体外转录和翻译系统、方法及合成物
<130> LHB2167528P
<150> 62/833,555
<151> 2019-04-12
<160> 136
<170> SIPOSequenceListing 1.0
<210> 1
<211> 219
<212> DNA
<213> Bacillaceae
<400> 1
atggccaaag atgatgtgat tgaagttgaa ggcaccgtta ttgaaaccct gccgaatgca 60
atgtttcgtg ttgaactgga aaatggtcat accgttctgg cacatgttag cggtaaaatt 120
cgcatgcact ttattcgtat tctgcctggt gatcgtgtta ccgttgaact gagcccgtac 180
gatctgaccc gtggtcgtat tacctatcgt tataaatga 219
<210> 2
<211> 72
<212> PRT
<213> Bacillaceae
<400> 2
Met Ala Lys Asp Asp Val Ile Glu Val Glu Gly Thr Val Ile Glu Thr
1 5 10 15
Leu Pro Asn Ala Met Phe Arg Val Glu Leu Glu Asn Gly His Thr Val
20 25 30
Leu Ala His Val Ser Gly Lys Ile Arg Met His Phe Ile Arg Ile Leu
35 40 45
Pro Gly Asp Arg Val Thr Val Glu Leu Ser Pro Tyr Asp Leu Thr Arg
50 55 60
Gly Arg Ile Thr Tyr Arg Tyr Lys
65 70
<210> 3
<211> 2220
<212> DNA
<213> Geobacillus stearothermophilus
<400> 3
atgagcaaaa tgcgcgttta tgagtacgcc aaaaaacaga atgttccgag caaagatgtg 60
atccacaaac tgaaagaaat gaacatcgaa gtgaacaacc atatggcaat gctggaagca 120
gatgttgttg aaaaactgga tcatcagtat cgtccgaata ccggcaaaaa agaagaaaaa 180
aaagccgaga agaaaaccga gaaaccgaaa cgtccgacac cagcaaaagc agcagatttt 240
gcagatgaag aaatcttcga tgatagcaaa gaagcagcca aaatgaaacc ggcaaagaaa 300
aaaggtgcac cgaaaggtaa agaaaccaaa aaaaccgaag cacagcagca agagaaaaaa 360
ctgctgcagg cagcgaaaaa gaaaggcaaa ggtccggcaa aagggaaaaa acaggcagca 420
ccggcagcca aacaggcacc gcagcctgcg aaaaaagaaa aagaactgcc gaaaaaaatc 480
acctttgaag gtagcctgac cgttgcagaa ctggcaaaaa aactgggtcg tgaaccgagc 540
gaaattatca aaaaactgtt tatgctgggt gtgatggcca ccattaatca ggatctggat 600
aaagatgcca ttgaactgat ttgcagcgat tatggtgttg aggttgaaga aaaagtgacc 660
atcgatgaaa ccaactttga agccattgaa attgttgatg caccggaaga tctggttgaa 720
cgtccgcctg ttgttaccat tatgggtcat gttgatcatg gtaaaaccac actgctggat 780
gcaattcgtc atagcaaagt taccgaacaa gaagcaggcg gtattacaca gcatattggt 840
gcatatcagg ttaccgtgaa cgataagaaa atcacgtttc tggatacacc gggtcatgaa 900
gcatttacca ccatgcgtgc acgtggtgca caggtgaccg atattgttat tctggttgtt 960
gcagcagatg atggcgttat gccgcagacc gttgaagcaa ttaatcatgc aaaagccgca 1020
aacgttccga ttattgttgc catcaacaaa atcgataaac cggaagcaaa tccggatcgt 1080
gttatgcaag aactgatgga atataatctg gttccggaag aatggggtgg tgataccatt 1140
ttttgtaaac tgagcgccaa aaccaaagaa ggtctggacc atctgctgga aatgattctg 1200
ctggttagcg aaatggaaga actgaaagcc aatccgaatc gtcgtgcagt tggcaccgtt 1260
attgaagcca aactggacaa aggtcgtggt ccggttgcga ccctgctgat tcaggcaggc 1320
accctgcgtg ttggtgatcc gattgttgtg ggcaccacct atggtcgtgt tcgtgcaatg 1380
gttaatgata gcggtcgtcg tgttaaagaa gcaaccccga gcatgccggt tgaaattacc 1440
ggtctgcatg aagttccgca ggcaggcgat cgttttatgg tttttgaaga tgagaaaaag 1500
gcacgccaga ttgccgaagc acgtgcacag cgtcagctgc aagaacagcg tagcgttaaa 1560
acccgtgtta gcctggatga cctgtttgag cagattaaac agggtgaaat gaaagagctg 1620
aacctgattg ttaaagccga tgttcagggt agcgttgaag ccctggttgc agcactgcag 1680
aaaattgatg ttgaaggtgt tcgcgtgaaa attatccatg cagccgttgg tgcaattacc 1740
gaaagcgata ttagcctggc aaccgcaagc aatgcaattg tgattggttt taatgttcgt 1800
ccggatgcaa atgcaaaacg tgcagcagaa agtgaaaaag tggatattcg tctgcaccgc 1860
attatctata acgtgatcga agaaattgag gcagccatga aaggtatgct ggatccggaa 1920
tatgaagaga aagttattgg tcaggcagaa gttcgtcaga cctttaaagt tagcaaagtg 1980
ggtacaattg ccggttgtta tgttaccgat ggtaaaatta cccgtgatag taaagttcgt 2040
ctgattcgtc agggtattgt tgtgtatgaa ggtgaaattg atagcctgaa acgctataaa 2100
gatgatgttc gtgaagttgc ccagggttat gaatgtggtc tgaccattaa aaacttcaac 2160
gacattaaag agggcgacgt tatcgaagcc tatatcatgc aagaagttgc acgcgcataa 2220
<210> 4
<211> 739
<212> PRT
<213> Geobacillus stearothermophilus
<400> 4
Met Ser Lys Met Arg Val Tyr Glu Tyr Ala Lys Lys Gln Asn Val Pro
1 5 10 15
Ser Lys Asp Val Ile His Lys Leu Lys Glu Met Asn Ile Glu Val Asn
20 25 30
Asn His Met Ala Met Leu Glu Ala Asp Val Val Glu Lys Leu Asp His
35 40 45
Gln Tyr Arg Pro Asn Thr Gly Lys Lys Glu Glu Lys Lys Ala Glu Lys
50 55 60
Lys Thr Glu Lys Pro Lys Arg Pro Thr Pro Ala Lys Ala Ala Asp Phe
65 70 75 80
Ala Asp Glu Glu Ile Phe Asp Asp Ser Lys Glu Ala Ala Lys Met Lys
85 90 95
Pro Ala Lys Lys Lys Gly Ala Pro Lys Gly Lys Glu Thr Lys Lys Thr
100 105 110
Glu Ala Gln Gln Gln Glu Lys Lys Leu Leu Gln Ala Ala Lys Lys Lys
115 120 125
Gly Lys Gly Pro Ala Lys Gly Lys Lys Gln Ala Ala Pro Ala Ala Lys
130 135 140
Gln Ala Pro Gln Pro Ala Lys Lys Glu Lys Glu Leu Pro Lys Lys Ile
145 150 155 160
Thr Phe Glu Gly Ser Leu Thr Val Ala Glu Leu Ala Lys Lys Leu Gly
165 170 175
Arg Glu Pro Ser Glu Ile Ile Lys Lys Leu Phe Met Leu Gly Val Met
180 185 190
Ala Thr Ile Asn Gln Asp Leu Asp Lys Asp Ala Ile Glu Leu Ile Cys
195 200 205
Ser Asp Tyr Gly Val Glu Val Glu Glu Lys Val Thr Ile Asp Glu Thr
210 215 220
Asn Phe Glu Ala Ile Glu Ile Val Asp Ala Pro Glu Asp Leu Val Glu
225 230 235 240
Arg Pro Pro Val Val Thr Ile Met Gly His Val Asp His Gly Lys Thr
245 250 255
Thr Leu Leu Asp Ala Ile Arg His Ser Lys Val Thr Glu Gln Glu Ala
260 265 270
Gly Gly Ile Thr Gln His Ile Gly Ala Tyr Gln Val Thr Val Asn Asp
275 280 285
Lys Lys Ile Thr Phe Leu Asp Thr Pro Gly His Glu Ala Phe Thr Thr
290 295 300
Met Arg Ala Arg Gly Ala Gln Val Thr Asp Ile Val Ile Leu Val Val
305 310 315 320
Ala Ala Asp Asp Gly Val Met Pro Gln Thr Val Glu Ala Ile Asn His
325 330 335
Ala Lys Ala Ala Asn Val Pro Ile Ile Val Ala Ile Asn Lys Ile Asp
340 345 350
Lys Pro Glu Ala Asn Pro Asp Arg Val Met Gln Glu Leu Met Glu Tyr
355 360 365
Asn Leu Val Pro Glu Glu Trp Gly Gly Asp Thr Ile Phe Cys Lys Leu
370 375 380
Ser Ala Lys Thr Lys Glu Gly Leu Asp His Leu Leu Glu Met Ile Leu
385 390 395 400
Leu Val Ser Glu Met Glu Glu Leu Lys Ala Asn Pro Asn Arg Arg Ala
405 410 415
Val Gly Thr Val Ile Glu Ala Lys Leu Asp Lys Gly Arg Gly Pro Val
420 425 430
Ala Thr Leu Leu Ile Gln Ala Gly Thr Leu Arg Val Gly Asp Pro Ile
435 440 445
Val Val Gly Thr Thr Tyr Gly Arg Val Arg Ala Met Val Asn Asp Ser
450 455 460
Gly Arg Arg Val Lys Glu Ala Thr Pro Ser Met Pro Val Glu Ile Thr
465 470 475 480
Gly Leu His Glu Val Pro Gln Ala Gly Asp Arg Phe Met Val Phe Glu
485 490 495
Asp Glu Lys Lys Ala Arg Gln Ile Ala Glu Ala Arg Ala Gln Arg Gln
500 505 510
Leu Gln Glu Gln Arg Ser Val Lys Thr Arg Val Ser Leu Asp Asp Leu
515 520 525
Phe Glu Gln Ile Lys Gln Gly Glu Met Lys Glu Leu Asn Leu Ile Val
530 535 540
Lys Ala Asp Val Gln Gly Ser Val Glu Ala Leu Val Ala Ala Leu Gln
545 550 555 560
Lys Ile Asp Val Glu Gly Val Arg Val Lys Ile Ile His Ala Ala Val
565 570 575
Gly Ala Ile Thr Glu Ser Asp Ile Ser Leu Ala Thr Ala Ser Asn Ala
580 585 590
Ile Val Ile Gly Phe Asn Val Arg Pro Asp Ala Asn Ala Lys Arg Ala
595 600 605
Ala Glu Ser Glu Lys Val Asp Ile Arg Leu His Arg Ile Ile Tyr Asn
610 615 620
Val Ile Glu Glu Ile Glu Ala Ala Met Lys Gly Met Leu Asp Pro Glu
625 630 635 640
Tyr Glu Glu Lys Val Ile Gly Gln Ala Glu Val Arg Gln Thr Phe Lys
645 650 655
Val Ser Lys Val Gly Thr Ile Ala Gly Cys Tyr Val Thr Asp Gly Lys
660 665 670
Ile Thr Arg Asp Ser Lys Val Arg Leu Ile Arg Gln Gly Ile Val Val
675 680 685
Tyr Glu Gly Glu Ile Asp Ser Leu Lys Arg Tyr Lys Asp Asp Val Arg
690 695 700
Glu Val Ala Gln Gly Tyr Glu Cys Gly Leu Thr Ile Lys Asn Phe Asn
705 710 715 720
Asp Ile Lys Glu Gly Asp Val Ile Glu Ala Tyr Ile Met Gln Glu Val
725 730 735
Ala Arg Ala
<210> 5
<211> 522
<212> DNA
<213> Geobacillus
<400> 5
atgatcagca aggactttat catcaatgag cagattcgtg cacgtgaagt tcgtctgatt 60
gatcagaatg gtgaacagct gggtatcaaa agcaaacaag aagcactgga aattgcagca 120
cgtcgtaatc tggatctggt tctggtggca ccgaatgcaa aaccgcctgt ttgtcgtatt 180
atggattatg gcaaatttcg cttcgagcag cagaaaaaag aaaaagaggc acgcaaaaag 240
cagaaagtga tcaatgttaa agaagtgcgt ctgagcccga ccattgaaga acatgatttt 300
aacaccaaac tgcgcaacgc acgcaaattt ctggaaaaag gtgataaagt gaaagccacc 360
attcgtttta aaggtcgtgc aatcacccat aaagaaattg gtcagcgtgt tctggatcgt 420
tttagcgaag catgtgcaga tattgcagtt gttgaaaccg caccgaaaat ggatggtcgt 480
aatatgtttc tggtgctggc tccgaaaaac gacaacaaat aa 522
<210> 6
<211> 173
<212> PRT
<213> Geobacillus
<400> 6
Met Ile Ser Lys Asp Phe Ile Ile Asn Glu Gln Ile Arg Ala Arg Glu
1 5 10 15
Val Arg Leu Ile Asp Gln Asn Gly Glu Gln Leu Gly Ile Lys Ser Lys
20 25 30
Gln Glu Ala Leu Glu Ile Ala Ala Arg Arg Asn Leu Asp Leu Val Leu
35 40 45
Val Ala Pro Asn Ala Lys Pro Pro Val Cys Arg Ile Met Asp Tyr Gly
50 55 60
Lys Phe Arg Phe Glu Gln Gln Lys Lys Glu Lys Glu Ala Arg Lys Lys
65 70 75 80
Gln Lys Val Ile Asn Val Lys Glu Val Arg Leu Ser Pro Thr Ile Glu
85 90 95
Glu His Asp Phe Asn Thr Lys Leu Arg Asn Ala Arg Lys Phe Leu Glu
100 105 110
Lys Gly Asp Lys Val Lys Ala Thr Ile Arg Phe Lys Gly Arg Ala Ile
115 120 125
Thr His Lys Glu Ile Gly Gln Arg Val Leu Asp Arg Phe Ser Glu Ala
130 135 140
Cys Ala Asp Ile Ala Val Val Glu Thr Ala Pro Lys Met Asp Gly Arg
145 150 155 160
Asn Met Phe Leu Val Leu Ala Pro Lys Asn Asp Asn Lys
165 170
<210> 7
<211> 2079
<212> DNA
<213> Geobacillus
<400> 7
atggcacgtg aattcagcct ggaaaaaacc cgtaatattg gtattatggc ccatatcgat 60
gcaggtaaaa ccaccaccac cgaacgtatt ctgttttata ccggtcgtgt gcataaaatt 120
ggtgaagttc atgaaggtgc agcaaccatg gattggatgg aacaagaaca agagcgtggt 180
attaccatta ccagcgcagc caccaccgca cagtggaaag gtcatcgtat taacattatt 240
gatacaccgg gtcacgttga ttttaccgtt gaagttgaac gtagcctgcg tgttctggat 300
ggtgcaatta ccgtgctgga tgcacagagc ggtgttgaac cgcagaccga aaccgtttgg 360
cgtcaggcaa ccacctatgg tgttccgcgt attgtttttg tgaacaagat ggataaaatc 420
ggtgccgatt tcctgtatag cgttaaaacc ctgcatgatc gtctgcaggc aaatgcacat 480
ccggttcagc tgccgattgg tgcagaagat cagtttagcg gtattattga tctggttgaa 540
atgtgcgcct atcactatca tgatgaactg ggcaaaaaca tcgaacgcat tgatattccg 600
gaagaatatc gtgatatggc cgaagagtat cacaacaaac tgattgaagc agttgcagaa 660
ctggatgaag aactgatgat gaaatatctg gaaggcgaag aaattaccgc agaggaactg 720
aaagcagcaa ttcgtaaagc aaccattagc gtggaatttt ttccggtttt ttgtggtagc 780
gccttcaaaa acaaaggtgt gcagctgctg ctggatggcg ttgttgatta tctgccgagt 840
ccggtggata ttcctgcaat tcgtggtgtt gttccggata ccgaagaaga agttacacgc 900
gaagcaagtg atgatgcacc gtttgcagca ctggccttta aaatcatgac cgatccgtat 960
gttggtaagc tgacctttat tcgtgtttat agcggcaccc tggatagcgg tagctatgtt 1020
atgaatacca ccaaaggtaa acgtgaacgt attggtcgtc tgctgcagat gcatgcaaat 1080
catcgtcaag aaatcagcaa agtttatgcc ggtgatattg cagcagcagt tggtctgaaa 1140
gataccacaa ccggtgatac cctgtgtgat gaaaaacatc cggtgattct ggaaagcatg 1200
cagtttccgg aaccggttat tagcgttgca attgaaccga aaagcaaagc cgatcaggat 1260
aaaatgagcc aggcactgca gaaactgcaa gaagaggatc cgacctttcg tgcacatacc 1320
gatccggaaa ccggtcagac cattattagt ggtatgggtg aactgcatct ggatatcatt 1380
gttgatcgta tgcgtcgcga atttaaagtt gaagcaaatg ttggtgcacc gcaggttgca 1440
tatcgtgaaa cctttcgtaa aagcgcacag gttgaaggca aatttatccg tcagagtggt 1500
ggtcgtggtc agtatggtca tgtttggatt gaattttcac cgaacgaacg cggtaaaggc 1560
tttgaatttg aaaatgcaat tgttggtggt gtggtgccga aagaatatgt tccggcagtt 1620
caggcaggtc tggaagaggc aatgcagaat ggtgttctgg caggttatcc ggttgttgat 1680
attaaagcca aactgttcga tggcagctat cacgatgttg atagcagcga aatggcattc 1740
aaaattgcag caagcctggc actgaaaaat gccgcaacca aatgtgatcc tgttctgctg 1800
gaaccgatta tgaaagtgga agttgttatc cctgaggaat atctgggtga tattatgggc 1860
gatattacca gccgtcgtgg tcgcattgaa ggtatggaag cacgtggtaa tgcccaggtt 1920
gttcgtgcaa tggttccgct ggcagaaatg tttggttatg caaccagcct gcgtagcaat 1980
acccaaggtc gtggcacctt tagcatggtt tttgatcatt atgaagaggt gcccaaaaac 2040
attgccgatg agatcatcaa aaaaaacaag ggcgaataa 2079
<210> 8
<211> 692
<212> PRT
<213> Geobacillus
<400> 8
Met Ala Arg Glu Phe Ser Leu Glu Lys Thr Arg Asn Ile Gly Ile Met
1 5 10 15
Ala His Ile Asp Ala Gly Lys Thr Thr Thr Thr Glu Arg Ile Leu Phe
20 25 30
Tyr Thr Gly Arg Val His Lys Ile Gly Glu Val His Glu Gly Ala Ala
35 40 45
Thr Met Asp Trp Met Glu Gln Glu Gln Glu Arg Gly Ile Thr Ile Thr
50 55 60
Ser Ala Ala Thr Thr Ala Gln Trp Lys Gly His Arg Ile Asn Ile Ile
65 70 75 80
Asp Thr Pro Gly His Val Asp Phe Thr Val Glu Val Glu Arg Ser Leu
85 90 95
Arg Val Leu Asp Gly Ala Ile Thr Val Leu Asp Ala Gln Ser Gly Val
100 105 110
Glu Pro Gln Thr Glu Thr Val Trp Arg Gln Ala Thr Thr Tyr Gly Val
115 120 125
Pro Arg Ile Val Phe Val Asn Lys Met Asp Lys Ile Gly Ala Asp Phe
130 135 140
Leu Tyr Ser Val Lys Thr Leu His Asp Arg Leu Gln Ala Asn Ala His
145 150 155 160
Pro Val Gln Leu Pro Ile Gly Ala Glu Asp Gln Phe Ser Gly Ile Ile
165 170 175
Asp Leu Val Glu Met Cys Ala Tyr His Tyr His Asp Glu Leu Gly Lys
180 185 190
Asn Ile Glu Arg Ile Asp Ile Pro Glu Glu Tyr Arg Asp Met Ala Glu
195 200 205
Glu Tyr His Asn Lys Leu Ile Glu Ala Val Ala Glu Leu Asp Glu Glu
210 215 220
Leu Met Met Lys Tyr Leu Glu Gly Glu Glu Ile Thr Ala Glu Glu Leu
225 230 235 240
Lys Ala Ala Ile Arg Lys Ala Thr Ile Ser Val Glu Phe Phe Pro Val
245 250 255
Phe Cys Gly Ser Ala Phe Lys Asn Lys Gly Val Gln Leu Leu Leu Asp
260 265 270
Gly Val Val Asp Tyr Leu Pro Ser Pro Val Asp Ile Pro Ala Ile Arg
275 280 285
Gly Val Val Pro Asp Thr Glu Glu Glu Val Thr Arg Glu Ala Ser Asp
290 295 300
Asp Ala Pro Phe Ala Ala Leu Ala Phe Lys Ile Met Thr Asp Pro Tyr
305 310 315 320
Val Gly Lys Leu Thr Phe Ile Arg Val Tyr Ser Gly Thr Leu Asp Ser
325 330 335
Gly Ser Tyr Val Met Asn Thr Thr Lys Gly Lys Arg Glu Arg Ile Gly
340 345 350
Arg Leu Leu Gln Met His Ala Asn His Arg Gln Glu Ile Ser Lys Val
355 360 365
Tyr Ala Gly Asp Ile Ala Ala Ala Val Gly Leu Lys Asp Thr Thr Thr
370 375 380
Gly Asp Thr Leu Cys Asp Glu Lys His Pro Val Ile Leu Glu Ser Met
385 390 395 400
Gln Phe Pro Glu Pro Val Ile Ser Val Ala Ile Glu Pro Lys Ser Lys
405 410 415
Ala Asp Gln Asp Lys Met Ser Gln Ala Leu Gln Lys Leu Gln Glu Glu
420 425 430
Asp Pro Thr Phe Arg Ala His Thr Asp Pro Glu Thr Gly Gln Thr Ile
435 440 445
Ile Ser Gly Met Gly Glu Leu His Leu Asp Ile Ile Val Asp Arg Met
450 455 460
Arg Arg Glu Phe Lys Val Glu Ala Asn Val Gly Ala Pro Gln Val Ala
465 470 475 480
Tyr Arg Glu Thr Phe Arg Lys Ser Ala Gln Val Glu Gly Lys Phe Ile
485 490 495
Arg Gln Ser Gly Gly Arg Gly Gln Tyr Gly His Val Trp Ile Glu Phe
500 505 510
Ser Pro Asn Glu Arg Gly Lys Gly Phe Glu Phe Glu Asn Ala Ile Val
515 520 525
Gly Gly Val Val Pro Lys Glu Tyr Val Pro Ala Val Gln Ala Gly Leu
530 535 540
Glu Glu Ala Met Gln Asn Gly Val Leu Ala Gly Tyr Pro Val Val Asp
545 550 555 560
Ile Lys Ala Lys Leu Phe Asp Gly Ser Tyr His Asp Val Asp Ser Ser
565 570 575
Glu Met Ala Phe Lys Ile Ala Ala Ser Leu Ala Leu Lys Asn Ala Ala
580 585 590
Thr Lys Cys Asp Pro Val Leu Leu Glu Pro Ile Met Lys Val Glu Val
595 600 605
Val Ile Pro Glu Glu Tyr Leu Gly Asp Ile Met Gly Asp Ile Thr Ser
610 615 620
Arg Arg Gly Arg Ile Glu Gly Met Glu Ala Arg Gly Asn Ala Gln Val
625 630 635 640
Val Arg Ala Met Val Pro Leu Ala Glu Met Phe Gly Tyr Ala Thr Ser
645 650 655
Leu Arg Ser Asn Thr Gln Gly Arg Gly Thr Phe Ser Met Val Phe Asp
660 665 670
His Tyr Glu Glu Val Pro Lys Asn Ile Ala Asp Glu Ile Ile Lys Lys
675 680 685
Asn Lys Gly Glu
690
<210> 9
<211> 1188
<212> DNA
<213> Geobacillus
<400> 9
atggccaaag ccaaatttga acgtaccaaa ccgcatgtta atattggcac cattggtcat 60
gttgatcatg gtaaaaccac actgaccgca gcaattacca ccgttctggc aaaacagggt 120
aaagccgaag caaaagcata tgatcagatt gatgcagcac cggaagaacg tgaacgtggt 180
attaccatta gcaccgcaca tgttgaatat gaaaccgatg cacgtcatta tgcccatgtt 240
gattgtccgg gtcatgcaga ttatgtgaaa aatatgatta ccggtgcagc acagatggat 300
ggtgcaattc tggttgttag cgcagcagat ggtccgatgc cgcagacacg tgaacatatt 360
ctgctgagcc gtcaggttgg tgttccgtat attgttgtgt ttctgaacaa atgcgatatg 420
gtggatgatg aagaactgct ggaactggtt gaaatggaag ttcgtgatct gctgtccgaa 480
tatgattttc cgggtgatga agttccggtt attaaaggta gcgcactgaa agcactggaa 540
ggtgatccgc agtgggaaga aaaaatcatt gaactgatga atgccgtgga tgagtatatt 600
ccgacaccgc agcgtgaagt tgataaaccg tttatgatgc cgatcgaaga tgtgtttagc 660
attaccggtc gtggcaccgt tgcaaccggt cgcgttgaac gtggcaccct gaaagttggt 720
gatccggttg aaattattgg tctgagtgat gaaccgaaaa ccaccaccgt taccggtgtt 780
gaaatgtttc gtaaactgtt agatcaggcc gaagccggtg ataatattgg tgcactgctg 840
cgtggtgttt cacgtgatga ggtggaacgt ggtcaggttc tggcgaaacc tggtagcatt 900
acaccgcata ccaaattcaa agcacaggtt tatgttctga ccaaagaaga aggcggtcgt 960
cataccccgt tttttagcaa ttatcgtccg cagttttatt tccgtaccac cgatgttacc 1020
ggtattatta ccctgccgga aggtgtggaa atggttatgc ctggtgataa cgttgaaatg 1080
accgtggaac tgattgcacc gattgcaatt gaagaaggca ccaaatttag cattcgtgaa 1140
ggtggtcgta ccgttggtgc aggtagcgtt agcgaaatta tcgaataa 1188
<210> 10
<211> 395
<212> PRT
<213> Geobacillus
<400> 10
Met Ala Lys Ala Lys Phe Glu Arg Thr Lys Pro His Val Asn Ile Gly
1 5 10 15
Thr Ile Gly His Val Asp His Gly Lys Thr Thr Leu Thr Ala Ala Ile
20 25 30
Thr Thr Val Leu Ala Lys Gln Gly Lys Ala Glu Ala Lys Ala Tyr Asp
35 40 45
Gln Ile Asp Ala Ala Pro Glu Glu Arg Glu Arg Gly Ile Thr Ile Ser
50 55 60
Thr Ala His Val Glu Tyr Glu Thr Asp Ala Arg His Tyr Ala His Val
65 70 75 80
Asp Cys Pro Gly His Ala Asp Tyr Val Lys Asn Met Ile Thr Gly Ala
85 90 95
Ala Gln Met Asp Gly Ala Ile Leu Val Val Ser Ala Ala Asp Gly Pro
100 105 110
Met Pro Gln Thr Arg Glu His Ile Leu Leu Ser Arg Gln Val Gly Val
115 120 125
Pro Tyr Ile Val Val Phe Leu Asn Lys Cys Asp Met Val Asp Asp Glu
130 135 140
Glu Leu Leu Glu Leu Val Glu Met Glu Val Arg Asp Leu Leu Ser Glu
145 150 155 160
Tyr Asp Phe Pro Gly Asp Glu Val Pro Val Ile Lys Gly Ser Ala Leu
165 170 175
Lys Ala Leu Glu Gly Asp Pro Gln Trp Glu Glu Lys Ile Ile Glu Leu
180 185 190
Met Asn Ala Val Asp Glu Tyr Ile Pro Thr Pro Gln Arg Glu Val Asp
195 200 205
Lys Pro Phe Met Met Pro Ile Glu Asp Val Phe Ser Ile Thr Gly Arg
210 215 220
Gly Thr Val Ala Thr Gly Arg Val Glu Arg Gly Thr Leu Lys Val Gly
225 230 235 240
Asp Pro Val Glu Ile Ile Gly Leu Ser Asp Glu Pro Lys Thr Thr Thr
245 250 255
Val Thr Gly Val Glu Met Phe Arg Lys Leu Leu Asp Gln Ala Glu Ala
260 265 270
Gly Asp Asn Ile Gly Ala Leu Leu Arg Gly Val Ser Arg Asp Glu Val
275 280 285
Glu Arg Gly Gln Val Leu Ala Lys Pro Gly Ser Ile Thr Pro His Thr
290 295 300
Lys Phe Lys Ala Gln Val Tyr Val Leu Thr Lys Glu Glu Gly Gly Arg
305 310 315 320
His Thr Pro Phe Phe Ser Asn Tyr Arg Pro Gln Phe Tyr Phe Arg Thr
325 330 335
Thr Asp Val Thr Gly Ile Ile Thr Leu Pro Glu Gly Val Glu Met Val
340 345 350
Met Pro Gly Asp Asn Val Glu Met Thr Val Glu Leu Ile Ala Pro Ile
355 360 365
Ala Ile Glu Glu Gly Thr Lys Phe Ser Ile Arg Glu Gly Gly Arg Thr
370 375 380
Val Gly Ala Gly Ser Val Ser Glu Ile Ile Glu
385 390 395
<210> 11
<211> 885
<212> DNA
<213> Geobacillus
<400> 11
atggcaatta ccgcacagat ggttaaagaa ctgcgtgaaa aaaccggtgc aggtatgatg 60
gattgtaaaa aagcactgac cgaaaccaat ggcgatatgg aaaaagcaat tgattggctg 120
cgcgaaaaag gtattgcaaa agcagcaaaa aaagccgatc gtattgcagc agaaggtatg 180
gcatatattg cagttgaagg taataccgca gttatcctgg aagttaatag cgaaaccgat 240
tttgtggcaa aaaacgaagc atttcagacc ctggtgaaag agctggcagc acatctgctg 300
aaacagaaac cggcaagcct ggatgaagca ctgggtcaga ccatggataa tggtagcacc 360
gttcaggatt atatcaatga agccattgcc aaaatcggcg aaaaaatcac cctgcgtcgt 420
tttgcagttg ttaataaagc agatggtgaa acctttggtg cctatctgca tatgggtggt 480
cgtattggtg ttctgaccct gctggcaggt aatgcaagcg aagatgttgc aaaagatgtg 540
gcaatgcata ttgcagccct gcatccgaaa tatgttagcc gtgatgatgt tccgcaagaa 600
gaaattgcac acgaacgtga agttctgaaa cagcaggcac tgaatgaagg caaaccggaa 660
aaaattgtgg aaaagatggt tgaaggtcgc ctgaacaaat tctatgaaga tgtttgtctg 720
ctggaacagg cctttgttaa aaatccggat gttaccgttc gtcagtatgt tgaaagcaat 780
ggtgccaccg ttaaacagtt tattcgttat gaagttggtg agggcttaga aaaacgccag 840
gataattttg ccgaagaagt tatgagccag gttcgcaaac agtaa 885
<210> 12
<211> 294
<212> PRT
<213> Geobacillus
<400> 12
Met Ala Ile Thr Ala Gln Met Val Lys Glu Leu Arg Glu Lys Thr Gly
1 5 10 15
Ala Gly Met Met Asp Cys Lys Lys Ala Leu Thr Glu Thr Asn Gly Asp
20 25 30
Met Glu Lys Ala Ile Asp Trp Leu Arg Glu Lys Gly Ile Ala Lys Ala
35 40 45
Ala Lys Lys Ala Asp Arg Ile Ala Ala Glu Gly Met Ala Tyr Ile Ala
50 55 60
Val Glu Gly Asn Thr Ala Val Ile Leu Glu Val Asn Ser Glu Thr Asp
65 70 75 80
Phe Val Ala Lys Asn Glu Ala Phe Gln Thr Leu Val Lys Glu Leu Ala
85 90 95
Ala His Leu Leu Lys Gln Lys Pro Ala Ser Leu Asp Glu Ala Leu Gly
100 105 110
Gln Thr Met Asp Asn Gly Ser Thr Val Gln Asp Tyr Ile Asn Glu Ala
115 120 125
Ile Ala Lys Ile Gly Glu Lys Ile Thr Leu Arg Arg Phe Ala Val Val
130 135 140
Asn Lys Ala Asp Gly Glu Thr Phe Gly Ala Tyr Leu His Met Gly Gly
145 150 155 160
Arg Ile Gly Val Leu Thr Leu Leu Ala Gly Asn Ala Ser Glu Asp Val
165 170 175
Ala Lys Asp Val Ala Met His Ile Ala Ala Leu His Pro Lys Tyr Val
180 185 190
Ser Arg Asp Asp Val Pro Gln Glu Glu Ile Ala His Glu Arg Glu Val
195 200 205
Leu Lys Gln Gln Ala Leu Asn Glu Gly Lys Pro Glu Lys Ile Val Glu
210 215 220
Lys Met Val Glu Gly Arg Leu Asn Lys Phe Tyr Glu Asp Val Cys Leu
225 230 235 240
Leu Glu Gln Ala Phe Val Lys Asn Pro Asp Val Thr Val Arg Gln Tyr
245 250 255
Val Glu Ser Asn Gly Ala Thr Val Lys Gln Phe Ile Arg Tyr Glu Val
260 265 270
Gly Glu Gly Leu Glu Lys Arg Gln Asp Asn Phe Ala Glu Glu Val Met
275 280 285
Ser Gln Val Arg Lys Gln
290
<210> 13
<211> 1827
<212> DNA
<213> Geobacillus
<400> 13
atgaaccgtg aggaacgtct gaaacgtcag gagcgtattc gtaacttcag catcattgcg 60
cacatcgacc acggtaaaag caccctggcg gatcgtatcc tggagaaaac cggtgcgctg 120
agcgagcgtg aactgcgtga acagaccctg gacatgatgg atctggagcg tgaacgtggt 180
atcaccatta agctgaacgc ggtgcaactg acctataagg cgaaaaacgg cgaggaatac 240
atcttccacc tgattgacac cccgggccac gtggatttta cctatgaagt tagccgtagc 300
ctggcggcgt gcgaaggtgc gattctggtg gttgatgcgg cgcagggtat tgaggcgcaa 360
accctggcga acgtgtacct ggcgattgac aacaacctgg aaatcctgcc ggttatcaac 420
aaaattgatc tgccgagcgc ggagccggaa cgtgtgcgtc aggagatcga agacgttatt 480
ggtctggatg cgagcgaggc ggtgctggcg agcgcgaagg ttggtatcgg cattgaggaa 540
atcctggagc aaattgtgga aaaaattccg gcgccgagcg gtgacccgga tgcgccgctg 600
aaggcgctga tctttgacag cctgtacgat ccgtatcgtg gcgtggttgc gtacgtgcgt 660
attgttgacg gtaccgttaa gccgggccag cgtatcaaaa tgatgagcac cggcaaggag 720
ttcgaagtga ccgaggtggg cgtttttacc ccgaagcaaa aaatcgttga cgaactgacc 780
gtgggtgatg ttggctatct gaccgcgagc attaagaacg tgaaagatac ccgtgttggt 840
gacaccatta ccgatgcgga gcgtccggcg gcggaaccgc tgccgggtta ccgtaaactg 900
aacccgatgg ttttctgcgg catgtatccg atcgacaccg cgcgttacaa cgatctgcgt 960
gaggcgctgg aaaagctgca gctgaacgac gcggcgctgc acttcgagcc ggaaaccagc 1020
caagcgctgg gtttcggctt tcgttgcggt tttctgggcc tgctgcacat ggagatcatt 1080
caggaacgta tcgagcgtga atttcacatc gatctgatta ccaccgcgcc gagcgtggtt 1140
tataaagtgc acctgaccga cggtaccgag gtgagcgttg ataacccgac caacatgccg 1200
gacccgcaaa aaatcgatcg tattgaggaa ccgtatgtga aggcgaccat tatggttccg 1260
aacgactacg tgggcccggt tatggaactg tgccagggta aacgtggcac cttcgtggac 1320
atgcaatacc tggatgagaa gcgtgttatg ctgatctatg acattccgct gagcgaaatc 1380
gtttacgact tctttgatgc gctgaagagc aacaccaaag gttacgcgag ctttgattat 1440
gagctgattg gctaccgtcc gagcaacctg gtgaaaatgg acatcctgct gaacggtgaa 1500
aagattgatg cgctgagctt catcgttcac cgtgaggcgg cgtatgaacg tggcaaagtg 1560
attgttgaga agctgaaaga cctgatcccg cgtcagcaat ttgaagtgcc ggttcaggcg 1620
gcgattggta acaaaatcat tgcgcgtagc accatcaagg cgctgcgtaa aaacgtgctg 1680
gcgaagtgct acggtggcga tgttagccgt aagcgtaaac tgctggagaa gcagaaagaa 1740
ggtaagaaac gtatgaaaca gattggtagc gttgaggtgc cgcaagaagc gttcatggcg 1800
gtgctgaaga tcgacgatca aaagaaa 1827
<210> 14
<211> 609
<212> PRT
<213> Geobacillus
<400> 14
Met Asn Arg Glu Glu Arg Leu Lys Arg Gln Glu Arg Ile Arg Asn Phe
1 5 10 15
Ser Ile Ile Ala His Ile Asp His Gly Lys Ser Thr Leu Ala Asp Arg
20 25 30
Ile Leu Glu Lys Thr Gly Ala Leu Ser Glu Arg Glu Leu Arg Glu Gln
35 40 45
Thr Leu Asp Met Met Asp Leu Glu Arg Glu Arg Gly Ile Thr Ile Lys
50 55 60
Leu Asn Ala Val Gln Leu Thr Tyr Lys Ala Lys Asn Gly Glu Glu Tyr
65 70 75 80
Ile Phe His Leu Ile Asp Thr Pro Gly His Val Asp Phe Thr Tyr Glu
85 90 95
Val Ser Arg Ser Leu Ala Ala Cys Glu Gly Ala Ile Leu Val Val Asp
100 105 110
Ala Ala Gln Gly Ile Glu Ala Gln Thr Leu Ala Asn Val Tyr Leu Ala
115 120 125
Ile Asp Asn Asn Leu Glu Ile Leu Pro Val Ile Asn Lys Ile Asp Leu
130 135 140
Pro Ser Ala Glu Pro Glu Arg Val Arg Gln Glu Ile Glu Asp Val Ile
145 150 155 160
Gly Leu Asp Ala Ser Glu Ala Val Leu Ala Ser Ala Lys Val Gly Ile
165 170 175
Gly Ile Glu Glu Ile Leu Glu Gln Ile Val Glu Lys Ile Pro Ala Pro
180 185 190
Ser Gly Asp Pro Asp Ala Pro Leu Lys Ala Leu Ile Phe Asp Ser Leu
195 200 205
Tyr Asp Pro Tyr Arg Gly Val Val Ala Tyr Val Arg Ile Val Asp Gly
210 215 220
Thr Val Lys Pro Gly Gln Arg Ile Lys Met Met Ser Thr Gly Lys Glu
225 230 235 240
Phe Glu Val Thr Glu Val Gly Val Phe Thr Pro Lys Gln Lys Ile Val
245 250 255
Asp Glu Leu Thr Val Gly Asp Val Gly Tyr Leu Thr Ala Ser Ile Lys
260 265 270
Asn Val Lys Asp Thr Arg Val Gly Asp Thr Ile Thr Asp Ala Glu Arg
275 280 285
Pro Ala Ala Glu Pro Leu Pro Gly Tyr Arg Lys Leu Asn Pro Met Val
290 295 300
Phe Cys Gly Met Tyr Pro Ile Asp Thr Ala Arg Tyr Asn Asp Leu Arg
305 310 315 320
Glu Ala Leu Glu Lys Leu Gln Leu Asn Asp Ala Ala Leu His Phe Glu
325 330 335
Pro Glu Thr Ser Gln Ala Leu Gly Phe Gly Phe Arg Cys Gly Phe Leu
340 345 350
Gly Leu Leu His Met Glu Ile Ile Gln Glu Arg Ile Glu Arg Glu Phe
355 360 365
His Ile Asp Leu Ile Thr Thr Ala Pro Ser Val Val Tyr Lys Val His
370 375 380
Leu Thr Asp Gly Thr Glu Val Ser Val Asp Asn Pro Thr Asn Met Pro
385 390 395 400
Asp Pro Gln Lys Ile Asp Arg Ile Glu Glu Pro Tyr Val Lys Ala Thr
405 410 415
Ile Met Val Pro Asn Asp Tyr Val Gly Pro Val Met Glu Leu Cys Gln
420 425 430
Gly Lys Arg Gly Thr Phe Val Asp Met Gln Tyr Leu Asp Glu Lys Arg
435 440 445
Val Met Leu Ile Tyr Asp Ile Pro Leu Ser Glu Ile Val Tyr Asp Phe
450 455 460
Phe Asp Ala Leu Lys Ser Asn Thr Lys Gly Tyr Ala Ser Phe Asp Tyr
465 470 475 480
Glu Leu Ile Gly Tyr Arg Pro Ser Asn Leu Val Lys Met Asp Ile Leu
485 490 495
Leu Asn Gly Glu Lys Ile Asp Ala Leu Ser Phe Ile Val His Arg Glu
500 505 510
Ala Ala Tyr Glu Arg Gly Lys Val Ile Val Glu Lys Leu Lys Asp Leu
515 520 525
Ile Pro Arg Gln Gln Phe Glu Val Pro Val Gln Ala Ala Ile Gly Asn
530 535 540
Lys Ile Ile Ala Arg Ser Thr Ile Lys Ala Leu Arg Lys Asn Val Leu
545 550 555 560
Ala Lys Cys Tyr Gly Gly Asp Val Ser Arg Lys Arg Lys Leu Leu Glu
565 570 575
Lys Gln Lys Glu Gly Lys Lys Arg Met Lys Gln Ile Gly Ser Val Glu
580 585 590
Val Pro Gln Glu Ala Phe Met Ala Val Leu Lys Ile Asp Asp Gln Lys
595 600 605
Lys
<210> 15
<211> 555
<212> DNA
<213> Geobacillus
<400> 15
atgatcagcg tgaacgactt ccgtaccggt ctgaccatcg aagttgatgg cgagatttgg 60
cgtgtgctgg aattccagca cgttaagccg ggtaaaggcg cggcgtttgt gcgtagcaag 120
ctgcgtaacc tgcgtaccgg tgcgatccaa gaacgtacct tccgtgcggg cgagaaggtg 180
aaccgtgcgc agattgacac ccgtaaaatg caatacctgt atgcgaacgg tgaccagcac 240
gtttttatgg atatggagac ctacgaacag atcgagctgc cggcgaaaca aattgagtat 300
gaactgaagt tcctgaaaga aaacatggaa gtgtttatca tgatgtacca aggtgaaacc 360
atcggcattg agctgccgaa caccgttgag ctgaaggtgg ttgagaccga accgggtatt 420
aaaggtgata ccgcgagcgg tggcagcaag ccggcgaaac tggaaaccgg cctggtggtt 480
caggtgccgt tctttgttaa cgagggtgac accctgatca ttaacaccgc ggatggcacc 540
tatgttagcc gtgcg 555
<210> 16
<211> 185
<212> PRT
<213> Geobacillus
<400> 16
Met Ile Ser Val Asn Asp Phe Arg Thr Gly Leu Thr Ile Glu Val Asp
1 5 10 15
Gly Glu Ile Trp Arg Val Leu Glu Phe Gln His Val Lys Pro Gly Lys
20 25 30
Gly Ala Ala Phe Val Arg Ser Lys Leu Arg Asn Leu Arg Thr Gly Ala
35 40 45
Ile Gln Glu Arg Thr Phe Arg Ala Gly Glu Lys Val Asn Arg Ala Gln
50 55 60
Ile Asp Thr Arg Lys Met Gln Tyr Leu Tyr Ala Asn Gly Asp Gln His
65 70 75 80
Val Phe Met Asp Met Glu Thr Tyr Glu Gln Ile Glu Leu Pro Ala Lys
85 90 95
Gln Ile Glu Tyr Glu Leu Lys Phe Leu Lys Glu Asn Met Glu Val Phe
100 105 110
Ile Met Met Tyr Gln Gly Glu Thr Ile Gly Ile Glu Leu Pro Asn Thr
115 120 125
Val Glu Leu Lys Val Val Glu Thr Glu Pro Gly Ile Lys Gly Asp Thr
130 135 140
Ala Ser Gly Gly Ser Lys Pro Ala Lys Leu Glu Thr Gly Leu Val Val
145 150 155 160
Gln Val Pro Phe Phe Val Asn Glu Gly Asp Thr Leu Ile Ile Asn Thr
165 170 175
Ala Asp Gly Thr Tyr Val Ser Arg Ala
180 185
<210> 17
<211> 1077
<212> DNA
<213> Geobacillus stearothermophilus
<400> 17
atgtttgatc gtctggaagc agttgaacag cgttatgaaa aactgaatga actgctgatg 60
gaaccggatg ttattaacga tccgaaaaaa ctgcgcgatt atagcaaaga acaggcagat 120
ctggaagaaa ccgttcagac ctatcgtgag tataaaagcg ttcgtgaaca gctggccgaa 180
gcaaaagcaa tgctggaaga gaaactggaa cctgaactgc gtgaaatggt gaaagaagaa 240
attggcgaac tggaagaacg tgaagaagca ctggttgaga aactgaaagt tctgctgctg 300
ccgaaagatc cgaatgatga aaaaaacgtg atcatggaaa ttcgtgcagc agccggtggc 360
gaagaagcag cactgtttgc cggtgatctg tatcgtatgt atacccgtta tgcagaaagc 420
caaggttgga aaaccgaagt tattgaagca agcccgaccg gtttaggtgg ttataaagaa 480
atcatcttca tgatcaatgg caagggtgca tacagcaaac tgaaatttga aaatggtgca 540
catcgtgttc agcgtgttcc ggaaaccgaa agcggtggtc gtattcatac cagcaccgca 600
accgttgcat gtctgccgga aatggaagaa atcgaagtgg aaatcaacga gaaagatatt 660
cgcgttgata cctttgcaag cagcggtcct ggtggtcaga gcgttaatac caccatgagc 720
gcagttcgtc tgacccatat tccgaccggt attgttgtta cctgtcagga tgaaaaatcc 780
cagatcaaaa acaaagaaaa agccatgaaa gtgctgcgtg cccgtatcta tgataaatat 840
cagcaagagg cacgtgcgga atatgatcag acccgtaaac aggcagttgg caccggtgat 900
cgtagcgaac gtattcgtac ctataacttt ccgcagaatc gtgttaccga tcatcgtatt 960
ggtctgacca ttcaaaaact ggatcaggtt ctggatggtc atctggatga aattatcgaa 1020
gcactgattc tggatgacca ggcaaaaaag ctggaacagg caaatgatgc aagctaa 1077
<210> 18
<211> 358
<212> PRT
<213> Geobacillus stearothermophilus
<400> 18
Met Phe Asp Arg Leu Glu Ala Val Glu Gln Arg Tyr Glu Lys Leu Asn
1 5 10 15
Glu Leu Leu Met Glu Pro Asp Val Ile Asn Asp Pro Lys Lys Leu Arg
20 25 30
Asp Tyr Ser Lys Glu Gln Ala Asp Leu Glu Glu Thr Val Gln Thr Tyr
35 40 45
Arg Glu Tyr Lys Ser Val Arg Glu Gln Leu Ala Glu Ala Lys Ala Met
50 55 60
Leu Glu Glu Lys Leu Glu Pro Glu Leu Arg Glu Met Val Lys Glu Glu
65 70 75 80
Ile Gly Glu Leu Glu Glu Arg Glu Glu Ala Leu Val Glu Lys Leu Lys
85 90 95
Val Leu Leu Leu Pro Lys Asp Pro Asn Asp Glu Lys Asn Val Ile Met
100 105 110
Glu Ile Arg Ala Ala Ala Gly Gly Glu Glu Ala Ala Leu Phe Ala Gly
115 120 125
Asp Leu Tyr Arg Met Tyr Thr Arg Tyr Ala Glu Ser Gln Gly Trp Lys
130 135 140
Thr Glu Val Ile Glu Ala Ser Pro Thr Gly Leu Gly Gly Tyr Lys Glu
145 150 155 160
Ile Ile Phe Met Ile Asn Gly Lys Gly Ala Tyr Ser Lys Leu Lys Phe
165 170 175
Glu Asn Gly Ala His Arg Val Gln Arg Val Pro Glu Thr Glu Ser Gly
180 185 190
Gly Arg Ile His Thr Ser Thr Ala Thr Val Ala Cys Leu Pro Glu Met
195 200 205
Glu Glu Ile Glu Val Glu Ile Asn Glu Lys Asp Ile Arg Val Asp Thr
210 215 220
Phe Ala Ser Ser Gly Pro Gly Gly Gln Ser Val Asn Thr Thr Met Ser
225 230 235 240
Ala Val Arg Leu Thr His Ile Pro Thr Gly Ile Val Val Thr Cys Gln
245 250 255
Asp Glu Lys Ser Gln Ile Lys Asn Lys Glu Lys Ala Met Lys Val Leu
260 265 270
Arg Ala Arg Ile Tyr Asp Lys Tyr Gln Gln Glu Ala Arg Ala Glu Tyr
275 280 285
Asp Gln Thr Arg Lys Gln Ala Val Gly Thr Gly Asp Arg Ser Glu Arg
290 295 300
Ile Arg Thr Tyr Asn Phe Pro Gln Asn Arg Val Thr Asp His Arg Ile
305 310 315 320
Gly Leu Thr Ile Gln Lys Leu Asp Gln Val Leu Asp Gly His Leu Asp
325 330 335
Glu Ile Ile Glu Ala Leu Ile Leu Asp Asp Gln Ala Lys Lys Leu Glu
340 345 350
Gln Ala Asn Asp Ala Ser
355
<210> 19
<211> 984
<212> DNA
<213> Geobacillus stearothermophilus
<400> 19
atggcagcac cgaatttttg ggatgatcag aaagcagcac aggcagttat tagcgaagca 60
aatgcactga aagatctggt ggaagaattt agcagcctgg aagaacgttt tgataatctg 120
gaagttacct acgaactgct gaaagaagaa ccggacgacg aactgcaggc agaactggtt 180
gaagaggcaa aaaaactgat gaaagatttt agcgaatttg aactgcagct gctgctgaat 240
gaaccgtatg atcaaaataa tgccatcctg gaactgcatc ctggtgccgg tggcaccgaa 300
agccaggatt gggcaagcat gctgctgcgt atgtataccc gttgggcaga aaaaaaaggc 360
tttaaagttg aaaccctgga ttatctgcct ggtgaagaag caggtattaa aagcgttacc 420
ctgctgatta aaggccataa tgcatatggt tatctgaaag ccgaaaaagg tgttcatcgt 480
ctggttcgta ttagcccgtt tgatgcaagc ggtcgtcgtc ataccagctt tgttagctgt 540
gaagttgtgc cggaactgga tgataacatt gaaattgaaa ttcgccctga agaactgaag 600
attgatacct atcgtagcag cggtgcaggc ggtcagcatg ttaataccac cgatagcgca 660
gtgcgtatta cccatctgcc gaccggtatt gttgttacct gtcagagcga acgtagccag 720
attaaaaacc gtgaaaaagc catgaatatg ctgaaagcca aactgtacca gaagaaatta 780
gaagaacagc aggccgagct ggccgaactg cgtggtgaac agaaagaaat tggttggggt 840
aatcagattc gcagctatgt ttttcatccg tacagcctgg ttaaagatca tcgtaccaat 900
gttgaagttg gtaatgttca ggccgttatg gatggtgaaa ttgatgtttt tatcgatgca 960
tacctgcgtg ccaaactgaa ataa 984
<210> 20
<211> 327
<212> PRT
<213> Geobacillus stearothermophilus
<400> 20
Met Ala Ala Pro Asn Phe Trp Asp Asp Gln Lys Ala Ala Gln Ala Val
1 5 10 15
Ile Ser Glu Ala Asn Ala Leu Lys Asp Leu Val Glu Glu Phe Ser Ser
20 25 30
Leu Glu Glu Arg Phe Asp Asn Leu Glu Val Thr Tyr Glu Leu Leu Lys
35 40 45
Glu Glu Pro Asp Asp Glu Leu Gln Ala Glu Leu Val Glu Glu Ala Lys
50 55 60
Lys Leu Met Lys Asp Phe Ser Glu Phe Glu Leu Gln Leu Leu Leu Asn
65 70 75 80
Glu Pro Tyr Asp Gln Asn Asn Ala Ile Leu Glu Leu His Pro Gly Ala
85 90 95
Gly Gly Thr Glu Ser Gln Asp Trp Ala Ser Met Leu Leu Arg Met Tyr
100 105 110
Thr Arg Trp Ala Glu Lys Lys Gly Phe Lys Val Glu Thr Leu Asp Tyr
115 120 125
Leu Pro Gly Glu Glu Ala Gly Ile Lys Ser Val Thr Leu Leu Ile Lys
130 135 140
Gly His Asn Ala Tyr Gly Tyr Leu Lys Ala Glu Lys Gly Val His Arg
145 150 155 160
Leu Val Arg Ile Ser Pro Phe Asp Ala Ser Gly Arg Arg His Thr Ser
165 170 175
Phe Val Ser Cys Glu Val Val Pro Glu Leu Asp Asp Asn Ile Glu Ile
180 185 190
Glu Ile Arg Pro Glu Glu Leu Lys Ile Asp Thr Tyr Arg Ser Ser Gly
195 200 205
Ala Gly Gly Gln His Val Asn Thr Thr Asp Ser Ala Val Arg Ile Thr
210 215 220
His Leu Pro Thr Gly Ile Val Val Thr Cys Gln Ser Glu Arg Ser Gln
225 230 235 240
Ile Lys Asn Arg Glu Lys Ala Met Asn Met Leu Lys Ala Lys Leu Tyr
245 250 255
Gln Lys Lys Leu Glu Glu Gln Gln Ala Glu Leu Ala Glu Leu Arg Gly
260 265 270
Glu Gln Lys Glu Ile Gly Trp Gly Asn Gln Ile Arg Ser Tyr Val Phe
275 280 285
His Pro Tyr Ser Leu Val Lys Asp His Arg Thr Asn Val Glu Val Gly
290 295 300
Asn Val Gln Ala Val Met Asp Gly Glu Ile Asp Val Phe Ile Asp Ala
305 310 315 320
Tyr Leu Arg Ala Lys Leu Lys
325
<210> 21
<211> 1578
<212> DNA
<213> Bacillus sp. X1
<400> 21
atgggtaacg atttcaagaa agaagtgctg agccgtcgta cctttgcgat cattagccat 60
ccggatgcgg gcaagaccac cctgaccgag aaactgctgc tgttcggtgg cgcgatccgt 120
gatgcgggta ccgttaaggc gaagaaaacc ggcaaatacg cgaccagcga ctggatggaa 180
atcgagaaac agcgtggtat tagcgtgacc agcagcgtta tgcaattcga ttacaacggt 240
tatcgtgtga acattctgga caccccgggc caccaggact ttagcgaaga tacctatcgt 300
accctgatgg cggtggacag cgcggttatg atcattgata gcgcgaaggg catcgaggac 360
caaaccatta agctgttcaa agtgtgccgt atgcgtggta tcccgatttt cacctttatc 420
aacaagctgg accgtcaggg caaacaaccg ctggagctgc tggcggaact ggaggaagtt 480
ctgggtatcg agagctaccc gatgaactgg ccgattggta tgggcaaaga atttctgggc 540
atctatgatc gttactataa ccgtattgag cagttccgtg tgaacgagga agagcgtttt 600
atcccgctga acgaagacgg tgaaattgag ggcaaccaca agctggttag cagcggtctg 660
tacgagcaga ccctggaaga gatcatgctg ctgaacgagg cgggtaacga atttagcgcg 720
gagcgtgtgg cggcgggtca actgaccccg gttttctttg gtagcgcgct gaccaacttc 780
ggcgtgcaga cctttctgga aacctatctg caatttgctc cgccgccgaa ggcgcgtaac 840
agcagcatcg gcgagattga tccgctgagc gaagagttta gcggcttcgt ttttaaaatt 900
caggcgaaca tgaacccggc gcaccgtgac cgtatcgcgt tcgtgcgtat ttgcagcggc 960
aagtttgagc gtggcatgag cgttaacctg ccgcgtctgg gcaagcagct gaaactgacc 1020
caaagcacca gcttcatggc ggaagagcgt aacaccgtgg aagaggcggt tagcggtgac 1080
atcattggcc tgtacgatac cggtacctat cagatcggcg ataccctgac cgtgggcaaa 1140
aacgacttcc agtttgagcg tctgccgcaa ttcaccccgg aactgtttgt gcgtgttagc 1200
gcgaagaacg ttatgcgtca gaagagcttt tacaaaggtc tgcaccagct ggtgcaagaa 1260
ggcgcgattc aactgtacaa gaccgttaaa accgatgagt atctgctggg tgcggtgggc 1320
cagctgcaat tcgaagtttt tgagcaccgt atgaagaacg aatataacgc ggaagtgctg 1380
atggaacgtc tgggtagcaa aatcgcgcgt tggattgaaa acgacgaggt tgatgaaaac 1440
ctgagcagca gccgtagcct gctggtgaaa gaccgttacg atcactatgt tttcctgttt 1500
gagaacgact tcgcgctgcg ttggtttcag gaaaagaacc cgaccatcaa actgtacaac 1560
ccgatggacc aacacgat 1578
<210> 22
<211> 526
<212> PRT
<213> Bacillus sp. X1
<400> 22
Met Gly Asn Asp Phe Lys Lys Glu Val Leu Ser Arg Arg Thr Phe Ala
1 5 10 15
Ile Ile Ser His Pro Asp Ala Gly Lys Thr Thr Leu Thr Glu Lys Leu
20 25 30
Leu Leu Phe Gly Gly Ala Ile Arg Asp Ala Gly Thr Val Lys Ala Lys
35 40 45
Lys Thr Gly Lys Tyr Ala Thr Ser Asp Trp Met Glu Ile Glu Lys Gln
50 55 60
Arg Gly Ile Ser Val Thr Ser Ser Val Met Gln Phe Asp Tyr Asn Gly
65 70 75 80
Tyr Arg Val Asn Ile Leu Asp Thr Pro Gly His Gln Asp Phe Ser Glu
85 90 95
Asp Thr Tyr Arg Thr Leu Met Ala Val Asp Ser Ala Val Met Ile Ile
100 105 110
Asp Ser Ala Lys Gly Ile Glu Asp Gln Thr Ile Lys Leu Phe Lys Val
115 120 125
Cys Arg Met Arg Gly Ile Pro Ile Phe Thr Phe Ile Asn Lys Leu Asp
130 135 140
Arg Gln Gly Lys Gln Pro Leu Glu Leu Leu Ala Glu Leu Glu Glu Val
145 150 155 160
Leu Gly Ile Glu Ser Tyr Pro Met Asn Trp Pro Ile Gly Met Gly Lys
165 170 175
Glu Phe Leu Gly Ile Tyr Asp Arg Tyr Tyr Asn Arg Ile Glu Gln Phe
180 185 190
Arg Val Asn Glu Glu Glu Arg Phe Ile Pro Leu Asn Glu Asp Gly Glu
195 200 205
Ile Glu Gly Asn His Lys Leu Val Ser Ser Gly Leu Tyr Glu Gln Thr
210 215 220
Leu Glu Glu Ile Met Leu Leu Asn Glu Ala Gly Asn Glu Phe Ser Ala
225 230 235 240
Glu Arg Val Ala Ala Gly Gln Leu Thr Pro Val Phe Phe Gly Ser Ala
245 250 255
Leu Thr Asn Phe Gly Val Gln Thr Phe Leu Glu Thr Tyr Leu Gln Phe
260 265 270
Ala Pro Pro Pro Lys Ala Arg Asn Ser Ser Ile Gly Glu Ile Asp Pro
275 280 285
Leu Ser Glu Glu Phe Ser Gly Phe Val Phe Lys Ile Gln Ala Asn Met
290 295 300
Asn Pro Ala His Arg Asp Arg Ile Ala Phe Val Arg Ile Cys Ser Gly
305 310 315 320
Lys Phe Glu Arg Gly Met Ser Val Asn Leu Pro Arg Leu Gly Lys Gln
325 330 335
Leu Lys Leu Thr Gln Ser Thr Ser Phe Met Ala Glu Glu Arg Asn Thr
340 345 350
Val Glu Glu Ala Val Ser Gly Asp Ile Ile Gly Leu Tyr Asp Thr Gly
355 360 365
Thr Tyr Gln Ile Gly Asp Thr Leu Thr Val Gly Lys Asn Asp Phe Gln
370 375 380
Phe Glu Arg Leu Pro Gln Phe Thr Pro Glu Leu Phe Val Arg Val Ser
385 390 395 400
Ala Lys Asn Val Met Arg Gln Lys Ser Phe Tyr Lys Gly Leu His Gln
405 410 415
Leu Val Gln Glu Gly Ala Ile Gln Leu Tyr Lys Thr Val Lys Thr Asp
420 425 430
Glu Tyr Leu Leu Gly Ala Val Gly Gln Leu Gln Phe Glu Val Phe Glu
435 440 445
His Arg Met Lys Asn Glu Tyr Asn Ala Glu Val Leu Met Glu Arg Leu
450 455 460
Gly Ser Lys Ile Ala Arg Trp Ile Glu Asn Asp Glu Val Asp Glu Asn
465 470 475 480
Leu Ser Ser Ser Arg Ser Leu Leu Val Lys Asp Arg Tyr Asp His Tyr
485 490 495
Val Phe Leu Phe Glu Asn Asp Phe Ala Leu Arg Trp Phe Gln Glu Lys
500 505 510
Asn Pro Thr Ile Lys Leu Tyr Asn Pro Met Asp Gln His Asp
515 520 525
<210> 23
<211> 558
<212> DNA
<213> Geobacillus
<400> 23
atggccaaac aggttattca gcaggccaaa gaaaaaatgg ataaagccgt tcaggcattt 60
acccgtgaac tggcaagcat tcgtgcaggt cgtgcaaatg caggtctgct ggaaaaagtt 120
accgttgatt attatggtgt tccgacgccg attaatcagc tggcgagcat tagcgttccg 180
gaagcacgtc tgctggtgat tcagccgtat gataaaagcg caatcaaaga gatggaaaaa 240
gcaattctgg caagcgatct gggtctgacc ccgagcaatg atggtagcgt tattcgtctg 300
gttattccgc ctctgaccga agaacgtcgt cgcgaactgg cgaaactggt gaaaaaatac 360
agcgaagatg caaaagttgc cgtgcgtaat attcgtcgtg atgcaaatga tgagctgaaa 420
aagctggaaa agaatggcga aattaccgaa gatgaactgc gtagctatac cgatgaagtt 480
cagaaactga ccgatgatca tatcgcaaaa attgacgcca tcaccaaaga gaaagaaaaa 540
gaagtcatgg aagtttaa 558
<210> 24
<211> 185
<212> PRT
<213> Geobacillus
<400> 24
Met Ala Lys Gln Val Ile Gln Gln Ala Lys Glu Lys Met Asp Lys Ala
1 5 10 15
Val Gln Ala Phe Thr Arg Glu Leu Ala Ser Ile Arg Ala Gly Arg Ala
20 25 30
Asn Ala Gly Leu Leu Glu Lys Val Thr Val Asp Tyr Tyr Gly Val Pro
35 40 45
Thr Pro Ile Asn Gln Leu Ala Ser Ile Ser Val Pro Glu Ala Arg Leu
50 55 60
Leu Val Ile Gln Pro Tyr Asp Lys Ser Ala Ile Lys Glu Met Glu Lys
65 70 75 80
Ala Ile Leu Ala Ser Asp Leu Gly Leu Thr Pro Ser Asn Asp Gly Ser
85 90 95
Val Ile Arg Leu Val Ile Pro Pro Leu Thr Glu Glu Arg Arg Arg Glu
100 105 110
Leu Ala Lys Leu Val Lys Lys Tyr Ser Glu Asp Ala Lys Val Ala Val
115 120 125
Arg Asn Ile Arg Arg Asp Ala Asn Asp Glu Leu Lys Lys Leu Glu Lys
130 135 140
Asn Gly Glu Ile Thr Glu Asp Glu Leu Arg Ser Tyr Thr Asp Glu Val
145 150 155 160
Gln Lys Leu Thr Asp Asp His Ile Ala Lys Ile Asp Ala Ile Thr Lys
165 170 175
Glu Lys Glu Lys Glu Val Met Glu Val
180 185
<210> 25
<211> 2637
<212> DNA
<213> Geobacillus stearothermophilus
<400> 25
atgaaaaaac tgaccagcgc acaggttcgt cgcatgtttc tggaattttt tcaagaaaaa 60
ggtcatgccg ttgaaccgag cgcaagcctg attccggttg atgatccgag cctgctgtgg 120
attaatagcg gtgttgcaac cctgaaaaaa tactttgatg gtcgtattgt tccggaaaat 180
ccgcgtattt gtaatgccca gaaaagcatt cgtaccaacg atattgaaaa tgtgggtaaa 240
accgcacgcc atcacacctt ttttgaaatg ctgggcaatt ttagcatcgg cgattatttc 300
aaacgtgaag caattcattg ggcctgggaa tttctgacca gtgataaatg gattggtttt 360
gatccggaac gtctgagcgt taccgttcat ccggaagatg aagaagcata taacatttgg 420
cgcaatgaaa ttggtctgcc ggaagaacgt attattcgtc tggaaggtaa cttttgggat 480
attggtgaag gtccgagcgg tccgaatacc gaaatctttt atgatcgtgg tgaagccttt 540
ggtaatgatc cgaatgatcc tgaactgtat ccaggtggtg aaaatgatcg ttatctggaa 600
gtttggaatc tggtgtttag ccagtttaat cataatccgg atggcaccta tacaccgctg 660
ccgaaaaaaa acattgatac cggcatgggt ttagaacgta tgtgtagcat tctgcaggat 720
gttccgacca attttgaaac cgacctgttt ctgccgatta ttcgtgcaac cgagcagatt 780
gccggtgaac gttatggtga agatccggat aaagatgttg cctttaaagt gattgccgat 840
catattcgcg cagttacctt tgcaattggt gatggtgcac tgccgagcaa tgaaggtcgt 900
ggttatgttc tgcgtcgtct gctgcgtcgt gcagttcgtt atgcaaaaca tattggtatt 960
gaacgtccgt tcatgtatga actggttccg gttgttggtg aaatcatgca cgattattat 1020
cccgaggtta aagagaaagc cgattttatt gcacgtgtga ttcgtaccga agaagaacgt 1080
tttcacgaaa ccctgcatga aggtctggca attctggcag aagttattga aaaagcaaaa 1140
gaacagggtt ccgatgttat tccgggtgaa gaggcatttc gtctgtatga tacctatggt 1200
tttccgattg aactgaccga agaatatgca gccgaagcag gtatgaccgt tgatcatgca 1260
ggttttgaac gtgaaatgga acgtcagcgt gaacgtgccc gtgcagcacg tcaggatgtt 1320
gatagtatgc aggttcaagg tggtgttctg ggtgatatta aagatgaaag tcgctttgtg 1380
ggctatgatg agctggttgc agcaagcacc gttattgcaa ttgttaaaga tggtcgtctg 1440
gtggaagaag ttaaagcagg cgaagaagca cagattattg ttgatgttac cccgttttat 1500
gcagaaagcg gtggtcagat tgcagatcag ggtgtttttg aaagcgaaac cggcaccgca 1560
gttgtgaaag atgttcagaa agcaccgaat ggtcagcatc tgcatgcaat tattgtggaa 1620
catggcaccg ttaaaaaagg tagccgttat accgcacgtg ttgatgaagc aaaacgtatg 1680
cgtattgtga aaaatcatac cgcaacacat ctgctgcatc aggcactgaa agacgttctg 1740
ggtcgtcatg ttaatcaggc aggtagcctg gttgcaccgg atcgtctgcg ttttgacttt 1800
acccattttg gtcaggttaa acccgaagaa ctggaacgta ttgaagcgat tgttaatgag 1860
cagatttgga aaagcctgcc ggtggatatt ttctataaac cgctggaaga ggcaaaagca 1920
atgggtgcaa tggcactgtt tggtgaaaaa tatggtgata ttgtgcgtgt ggttaaagtg 1980
ggtgattata gcctggaact gtgtggtggt tgtcatgtgc cgaataccag cgccattggt 2040
ctgtttaaaa tcgttagcga aagcggtatt ggtgcaggca cccgtcgcat tgaagcagtt 2100
accggtgaag cagcatatcg ttttatgagc gaacagctgg ccattctgca agaagcagca 2160
cagaaactga aaaccagtcc gaaagaactg aatgcacgtc tggatggcct gtttgcagaa 2220
ctgaaagaat tagaacgcga aaatgaaagc ctggcagccc gtctggcaca tatggaagca 2280
gaacatctga cccgtcaggt aaaagatgtt aatggtgttc cggttctggc agcaaaagtt 2340
caggcaaatg atatgaatca gctgcgtgcc atggccgatg atctgaaaca aaaactgggt 2400
acagcagtta ttgttctggc aagcgcacaa ggtggtaaag ttcagctgat tgcagccgtt 2460
acagatgacc tggtaaaaaa aggttttcat gcgggtaaac tggttaaaga agttgcaagc 2520
cgttgcggtg gtggtggcgg tggtcgtccg gatctggcac aggcaggcgg taaagatccg 2580
agcaaagttg gtgaagcact gggttatgtt gaaacctggg ttaaaagcgt gagctaa 2637
<210> 26
<211> 878
<212> PRT
<213> Geobacillus stearothermophilus
<400> 26
Met Lys Lys Leu Thr Ser Ala Gln Val Arg Arg Met Phe Leu Glu Phe
1 5 10 15
Phe Gln Glu Lys Gly His Ala Val Glu Pro Ser Ala Ser Leu Ile Pro
20 25 30
Val Asp Asp Pro Ser Leu Leu Trp Ile Asn Ser Gly Val Ala Thr Leu
35 40 45
Lys Lys Tyr Phe Asp Gly Arg Ile Val Pro Glu Asn Pro Arg Ile Cys
50 55 60
Asn Ala Gln Lys Ser Ile Arg Thr Asn Asp Ile Glu Asn Val Gly Lys
65 70 75 80
Thr Ala Arg His His Thr Phe Phe Glu Met Leu Gly Asn Phe Ser Ile
85 90 95
Gly Asp Tyr Phe Lys Arg Glu Ala Ile His Trp Ala Trp Glu Phe Leu
100 105 110
Thr Ser Asp Lys Trp Ile Gly Phe Asp Pro Glu Arg Leu Ser Val Thr
115 120 125
Val His Pro Glu Asp Glu Glu Ala Tyr Asn Ile Trp Arg Asn Glu Ile
130 135 140
Gly Leu Pro Glu Glu Arg Ile Ile Arg Leu Glu Gly Asn Phe Trp Asp
145 150 155 160
Ile Gly Glu Gly Pro Ser Gly Pro Asn Thr Glu Ile Phe Tyr Asp Arg
165 170 175
Gly Glu Ala Phe Gly Asn Asp Pro Asn Asp Pro Glu Leu Tyr Pro Gly
180 185 190
Gly Glu Asn Asp Arg Tyr Leu Glu Val Trp Asn Leu Val Phe Ser Gln
195 200 205
Phe Asn His Asn Pro Asp Gly Thr Tyr Thr Pro Leu Pro Lys Lys Asn
210 215 220
Ile Asp Thr Gly Met Gly Leu Glu Arg Met Cys Ser Ile Leu Gln Asp
225 230 235 240
Val Pro Thr Asn Phe Glu Thr Asp Leu Phe Leu Pro Ile Ile Arg Ala
245 250 255
Thr Glu Gln Ile Ala Gly Glu Arg Tyr Gly Glu Asp Pro Asp Lys Asp
260 265 270
Val Ala Phe Lys Val Ile Ala Asp His Ile Arg Ala Val Thr Phe Ala
275 280 285
Ile Gly Asp Gly Ala Leu Pro Ser Asn Glu Gly Arg Gly Tyr Val Leu
290 295 300
Arg Arg Leu Leu Arg Arg Ala Val Arg Tyr Ala Lys His Ile Gly Ile
305 310 315 320
Glu Arg Pro Phe Met Tyr Glu Leu Val Pro Val Val Gly Glu Ile Met
325 330 335
His Asp Tyr Tyr Pro Glu Val Lys Glu Lys Ala Asp Phe Ile Ala Arg
340 345 350
Val Ile Arg Thr Glu Glu Glu Arg Phe His Glu Thr Leu His Glu Gly
355 360 365
Leu Ala Ile Leu Ala Glu Val Ile Glu Lys Ala Lys Glu Gln Gly Ser
370 375 380
Asp Val Ile Pro Gly Glu Glu Ala Phe Arg Leu Tyr Asp Thr Tyr Gly
385 390 395 400
Phe Pro Ile Glu Leu Thr Glu Glu Tyr Ala Ala Glu Ala Gly Met Thr
405 410 415
Val Asp His Ala Gly Phe Glu Arg Glu Met Glu Arg Gln Arg Glu Arg
420 425 430
Ala Arg Ala Ala Arg Gln Asp Val Asp Ser Met Gln Val Gln Gly Gly
435 440 445
Val Leu Gly Asp Ile Lys Asp Glu Ser Arg Phe Val Gly Tyr Asp Glu
450 455 460
Leu Val Ala Ala Ser Thr Val Ile Ala Ile Val Lys Asp Gly Arg Leu
465 470 475 480
Val Glu Glu Val Lys Ala Gly Glu Glu Ala Gln Ile Ile Val Asp Val
485 490 495
Thr Pro Phe Tyr Ala Glu Ser Gly Gly Gln Ile Ala Asp Gln Gly Val
500 505 510
Phe Glu Ser Glu Thr Gly Thr Ala Val Val Lys Asp Val Gln Lys Ala
515 520 525
Pro Asn Gly Gln His Leu His Ala Ile Ile Val Glu His Gly Thr Val
530 535 540
Lys Lys Gly Ser Arg Tyr Thr Ala Arg Val Asp Glu Ala Lys Arg Met
545 550 555 560
Arg Ile Val Lys Asn His Thr Ala Thr His Leu Leu His Gln Ala Leu
565 570 575
Lys Asp Val Leu Gly Arg His Val Asn Gln Ala Gly Ser Leu Val Ala
580 585 590
Pro Asp Arg Leu Arg Phe Asp Phe Thr His Phe Gly Gln Val Lys Pro
595 600 605
Glu Glu Leu Glu Arg Ile Glu Ala Ile Val Asn Glu Gln Ile Trp Lys
610 615 620
Ser Leu Pro Val Asp Ile Phe Tyr Lys Pro Leu Glu Glu Ala Lys Ala
625 630 635 640
Met Gly Ala Met Ala Leu Phe Gly Glu Lys Tyr Gly Asp Ile Val Arg
645 650 655
Val Val Lys Val Gly Asp Tyr Ser Leu Glu Leu Cys Gly Gly Cys His
660 665 670
Val Pro Asn Thr Ser Ala Ile Gly Leu Phe Lys Ile Val Ser Glu Ser
675 680 685
Gly Ile Gly Ala Gly Thr Arg Arg Ile Glu Ala Val Thr Gly Glu Ala
690 695 700
Ala Tyr Arg Phe Met Ser Glu Gln Leu Ala Ile Leu Gln Glu Ala Ala
705 710 715 720
Gln Lys Leu Lys Thr Ser Pro Lys Glu Leu Asn Ala Arg Leu Asp Gly
725 730 735
Leu Phe Ala Glu Leu Lys Glu Leu Glu Arg Glu Asn Glu Ser Leu Ala
740 745 750
Ala Arg Leu Ala His Met Glu Ala Glu His Leu Thr Arg Gln Val Lys
755 760 765
Asp Val Asn Gly Val Pro Val Leu Ala Ala Lys Val Gln Ala Asn Asp
770 775 780
Met Asn Gln Leu Arg Ala Met Ala Asp Asp Leu Lys Gln Lys Leu Gly
785 790 795 800
Thr Ala Val Ile Val Leu Ala Ser Ala Gln Gly Gly Lys Val Gln Leu
805 810 815
Ile Ala Ala Val Thr Asp Asp Leu Val Lys Lys Gly Phe His Ala Gly
820 825 830
Lys Leu Val Lys Glu Val Ala Ser Arg Cys Gly Gly Gly Gly Gly Gly
835 840 845
Arg Pro Asp Leu Ala Gln Ala Gly Gly Lys Asp Pro Ser Lys Val Gly
850 855 860
Glu Ala Leu Gly Tyr Val Glu Thr Trp Val Lys Ser Val Ser
865 870 875
<210> 27
<211> 1674
<212> DNA
<213> Geobacillus
<400> 27
atgaatattg tgggccagat caaagaaaaa atgaaagaag aaattcgtca ggcagcagtt 60
cgtgcaggtc tggcaagcgc agatgaactg ccggatgttc tgctggaagt tccgcgtgat 120
aaagcacatg gtgattatag caccaatatt gcaatgcagc tggcacgtat tgcaaaaaaa 180
ccgcctcgtg caattgccga agcaattgtt ggtcagctgg atcgtgaacg tatgagcgtt 240
gcccgtattg aaattgcagg tccgggtttt atcaacttct atatggataa tcgttacctg 300
accgcagttg ttccggcaat tctgcaggca ggtcaggcat atggtgaaag taatgttggt 360
aatggtgaga aagtccaggt tgaatttgtt agcgcaaatc cgaccggtga tctgcatctg 420
ggtcatgcac gtggtgcagc agttggtgat agcctgtgta atattctggc aaaagcaggt 480
tttgatgtga cccgtgaata ctatattaat gatgcaggca agcagatcta caatctggcc 540
aaaagcgttg aagcacgtta ttttcaggca ctgggtgttg atatgccgct gccggaagat 600
ggttattatg gtgatgatat tgtggaaatc ggcaaaaaac tggccgaaga atatggtgat 660
cgtttcgttg aaatggaaga agaggaacgt ctggcatttt ttcgtgatta tggtctgcgt 720
tatgagctgg aaaaaatcaa aaaagatctg gccgattttc gcgttccgtt tgatgtttgg 780
tatagcgaaa ccagcctgta tgaaagcggt aaaattgatg aagcactgag caccctgcgt 840
gaacgtggtt atatctatga acaggatggt gcaacctggt ttcgtagcac cgcatttgga 900
gatgataaag atcgtgttct gattaaacag gacggcacct atacctatct gctgccggat 960
attgcatatc atcaggataa actgcgtcgc ggttttaaga aactgattaa catttggggt 1020
gccgatcatc atggttatat tcctcgcatg aaagcagcaa ttgcagcact gggttatgat 1080
ccggaagcac tggaagttga aattattcag atggtgaatc tgtatcagaa tggcgaacgt 1140
gtgaaaatga gcaaacgtac cggtaaagca gttaccatgc gtgaactgat ggaagaggtt 1200
ggtgttgatg cagttcgtta tttctttgca atgcgtagcg gtgataccca tctggatttt 1260
gatatggatc tggcagttag ccagagcaat gaaaatccgg tttattatgt tcagtatgcc 1320
catgcgcgtg ttagcagcat tctgcgtcag gcggaagaac agcatattag ctatgatggt 1380
gatctggcac tgcatcatct ggttgaaacc gaaaaagaaa ttgagctgct gaaagtgctg 1440
ggtgattttc cggatgttgt tgcagaagca gcactgaaac gtatgccgca tcgtgttacc 1500
gcatatgcat ttgacctggc cagcgcactg catagctttt ataacgccga aaaagttctg 1560
gatctggaca acatcgaaaa aaccaaagca cgtctggccc tggttaaagc cgttcagatt 1620
acactgcaga atgcactggc cctgattggt gtgagcgcac cggaacaaat gtaa 1674
<210> 28
<211> 557
<212> PRT
<213> Geobacillus
<400> 28
Met Asn Ile Val Gly Gln Ile Lys Glu Lys Met Lys Glu Glu Ile Arg
1 5 10 15
Gln Ala Ala Val Arg Ala Gly Leu Ala Ser Ala Asp Glu Leu Pro Asp
20 25 30
Val Leu Leu Glu Val Pro Arg Asp Lys Ala His Gly Asp Tyr Ser Thr
35 40 45
Asn Ile Ala Met Gln Leu Ala Arg Ile Ala Lys Lys Pro Pro Arg Ala
50 55 60
Ile Ala Glu Ala Ile Val Gly Gln Leu Asp Arg Glu Arg Met Ser Val
65 70 75 80
Ala Arg Ile Glu Ile Ala Gly Pro Gly Phe Ile Asn Phe Tyr Met Asp
85 90 95
Asn Arg Tyr Leu Thr Ala Val Val Pro Ala Ile Leu Gln Ala Gly Gln
100 105 110
Ala Tyr Gly Glu Ser Asn Val Gly Asn Gly Glu Lys Val Gln Val Glu
115 120 125
Phe Val Ser Ala Asn Pro Thr Gly Asp Leu His Leu Gly His Ala Arg
130 135 140
Gly Ala Ala Val Gly Asp Ser Leu Cys Asn Ile Leu Ala Lys Ala Gly
145 150 155 160
Phe Asp Val Thr Arg Glu Tyr Tyr Ile Asn Asp Ala Gly Lys Gln Ile
165 170 175
Tyr Asn Leu Ala Lys Ser Val Glu Ala Arg Tyr Phe Gln Ala Leu Gly
180 185 190
Val Asp Met Pro Leu Pro Glu Asp Gly Tyr Tyr Gly Asp Asp Ile Val
195 200 205
Glu Ile Gly Lys Lys Leu Ala Glu Glu Tyr Gly Asp Arg Phe Val Glu
210 215 220
Met Glu Glu Glu Glu Arg Leu Ala Phe Phe Arg Asp Tyr Gly Leu Arg
225 230 235 240
Tyr Glu Leu Glu Lys Ile Lys Lys Asp Leu Ala Asp Phe Arg Val Pro
245 250 255
Phe Asp Val Trp Tyr Ser Glu Thr Ser Leu Tyr Glu Ser Gly Lys Ile
260 265 270
Asp Glu Ala Leu Ser Thr Leu Arg Glu Arg Gly Tyr Ile Tyr Glu Gln
275 280 285
Asp Gly Ala Thr Trp Phe Arg Ser Thr Ala Phe Gly Asp Asp Lys Asp
290 295 300
Arg Val Leu Ile Lys Gln Asp Gly Thr Tyr Thr Tyr Leu Leu Pro Asp
305 310 315 320
Ile Ala Tyr His Gln Asp Lys Leu Arg Arg Gly Phe Lys Lys Leu Ile
325 330 335
Asn Ile Trp Gly Ala Asp His His Gly Tyr Ile Pro Arg Met Lys Ala
340 345 350
Ala Ile Ala Ala Leu Gly Tyr Asp Pro Glu Ala Leu Glu Val Glu Ile
355 360 365
Ile Gln Met Val Asn Leu Tyr Gln Asn Gly Glu Arg Val Lys Met Ser
370 375 380
Lys Arg Thr Gly Lys Ala Val Thr Met Arg Glu Leu Met Glu Glu Val
385 390 395 400
Gly Val Asp Ala Val Arg Tyr Phe Phe Ala Met Arg Ser Gly Asp Thr
405 410 415
His Leu Asp Phe Asp Met Asp Leu Ala Val Ser Gln Ser Asn Glu Asn
420 425 430
Pro Val Tyr Tyr Val Gln Tyr Ala His Ala Arg Val Ser Ser Ile Leu
435 440 445
Arg Gln Ala Glu Glu Gln His Ile Ser Tyr Asp Gly Asp Leu Ala Leu
450 455 460
His His Leu Val Glu Thr Glu Lys Glu Ile Glu Leu Leu Lys Val Leu
465 470 475 480
Gly Asp Phe Pro Asp Val Val Ala Glu Ala Ala Leu Lys Arg Met Pro
485 490 495
His Arg Val Thr Ala Tyr Ala Phe Asp Leu Ala Ser Ala Leu His Ser
500 505 510
Phe Tyr Asn Ala Glu Lys Val Leu Asp Leu Asp Asn Ile Glu Lys Thr
515 520 525
Lys Ala Arg Leu Ala Leu Val Lys Ala Val Gln Ile Thr Leu Gln Asn
530 535 540
Ala Leu Ala Leu Ile Gly Val Ser Ala Pro Glu Gln Met
545 550 555
<210> 29
<211> 1329
<212> DNA
<213> Geobacillus
<400> 29
atggatgtga gcattattgg tggtaatcag tgtgttaaaa ccaccaccat tgccgaagtt 60
aatcagtatg ttggtcagca ggttaccatt ggtgcatggc tggcaaataa acgtagcagc 120
ggtaaaattg tttttctgca gctgcgtgat ggcaccggtt ttattcaggg tgttgttgaa 180
aaagccaatg ttagcgaaga ggtttttcag cgtgcaaaaa ccctgacaca agaaaccagc 240
ctgtatgtga ccggcaccgt tcgtattgat gaacgtagcc cgtttggtta tgaactgagc 300
gttgccgatc tgcaggttat tcaagaagca gttgattatc cgattacgcc gaaagaacat 360
ggtgttgaat ttctgatgga tcatcgtcat ctgtggctgc gtagccgtcg tcagcatgca 420
attatgaaaa ttcgcaacga aattatccgt gccacctatg aatttttcaa cgatcgtggt 480
tttgtgaaag tggatgcacc gattctgacc ggtagcgcac cggaaggcac caccgaactg 540
tttcatacca aatatttcga tgaggatgca tatctgagcc agagcggtca gctgtatatg 600
gaagcagcag caatggcact gggtaaagtt tttagctttg gtccgacctt tcgtgccgaa 660
aaaagcaaaa cccgtcgcca tctgattgaa ttttggatgg ttgaaccgga aatggccttt 720
tatgaatttg aagataatct gcgcctgcaa gaggaatatg ttagctatct ggttcagagc 780
gttctggaac gttgtcgtct ggaactgggt cgcctgggtc gtgatgttag caaactggaa 840
ttagttaaac cgccttttcc gcgtctgacc tatgatgaag caattaaact gctgcatgaa 900
aaaggcctga ccgatattga atggggtgat gattttggtg caccgcatga aaccgcaatt 960
gcagaaagct ttgataaacc ggtgtttatc acccattatc cgaccagcct gaaaccgttt 1020
tatatgcagc cggatccgaa tcgtccggat gttgttctgt gtgcagatct gattgctccg 1080
gaaggttatg gtgaaattat tggcggtagc gaacgcatcc atgattatga gctgctgaaa 1140
cgtcgcctgg aagaacatca tctgccgctg gaagcatatg aatggtatct ggatctgcgt 1200
aaatatggta gcgttccgca tagcggtttt ggtctgggtt tagaacgtac cgttgcatgg 1260
atttgcggtg ttgaacatgt gcgtgaaacc attccgtttc cacgtctgct gaatcgtctg 1320
tatccgtaa 1329
<210> 30
<211> 442
<212> PRT
<213> Geobacillus
<400> 30
Met Asp Val Ser Ile Ile Gly Gly Asn Gln Cys Val Lys Thr Thr Thr
1 5 10 15
Ile Ala Glu Val Asn Gln Tyr Val Gly Gln Gln Val Thr Ile Gly Ala
20 25 30
Trp Leu Ala Asn Lys Arg Ser Ser Gly Lys Ile Val Phe Leu Gln Leu
35 40 45
Arg Asp Gly Thr Gly Phe Ile Gln Gly Val Val Glu Lys Ala Asn Val
50 55 60
Ser Glu Glu Val Phe Gln Arg Ala Lys Thr Leu Thr Gln Glu Thr Ser
65 70 75 80
Leu Tyr Val Thr Gly Thr Val Arg Ile Asp Glu Arg Ser Pro Phe Gly
85 90 95
Tyr Glu Leu Ser Val Ala Asp Leu Gln Val Ile Gln Glu Ala Val Asp
100 105 110
Tyr Pro Ile Thr Pro Lys Glu His Gly Val Glu Phe Leu Met Asp His
115 120 125
Arg His Leu Trp Leu Arg Ser Arg Arg Gln His Ala Ile Met Lys Ile
130 135 140
Arg Asn Glu Ile Ile Arg Ala Thr Tyr Glu Phe Phe Asn Asp Arg Gly
145 150 155 160
Phe Val Lys Val Asp Ala Pro Ile Leu Thr Gly Ser Ala Pro Glu Gly
165 170 175
Thr Thr Glu Leu Phe His Thr Lys Tyr Phe Asp Glu Asp Ala Tyr Leu
180 185 190
Ser Gln Ser Gly Gln Leu Tyr Met Glu Ala Ala Ala Met Ala Leu Gly
195 200 205
Lys Val Phe Ser Phe Gly Pro Thr Phe Arg Ala Glu Lys Ser Lys Thr
210 215 220
Arg Arg His Leu Ile Glu Phe Trp Met Val Glu Pro Glu Met Ala Phe
225 230 235 240
Tyr Glu Phe Glu Asp Asn Leu Arg Leu Gln Glu Glu Tyr Val Ser Tyr
245 250 255
Leu Val Gln Ser Val Leu Glu Arg Cys Arg Leu Glu Leu Gly Arg Leu
260 265 270
Gly Arg Asp Val Ser Lys Leu Glu Leu Val Lys Pro Pro Phe Pro Arg
275 280 285
Leu Thr Tyr Asp Glu Ala Ile Lys Leu Leu His Glu Lys Gly Leu Thr
290 295 300
Asp Ile Glu Trp Gly Asp Asp Phe Gly Ala Pro His Glu Thr Ala Ile
305 310 315 320
Ala Glu Ser Phe Asp Lys Pro Val Phe Ile Thr His Tyr Pro Thr Ser
325 330 335
Leu Lys Pro Phe Tyr Met Gln Pro Asp Pro Asn Arg Pro Asp Val Val
340 345 350
Leu Cys Ala Asp Leu Ile Ala Pro Glu Gly Tyr Gly Glu Ile Ile Gly
355 360 365
Gly Ser Glu Arg Ile His Asp Tyr Glu Leu Leu Lys Arg Arg Leu Glu
370 375 380
Glu His His Leu Pro Leu Glu Ala Tyr Glu Trp Tyr Leu Asp Leu Arg
385 390 395 400
Lys Tyr Gly Ser Val Pro His Ser Gly Phe Gly Leu Gly Leu Glu Arg
405 410 415
Thr Val Ala Trp Ile Cys Gly Val Glu His Val Arg Glu Thr Ile Pro
420 425 430
Phe Pro Arg Leu Leu Asn Arg Leu Tyr Pro
435 440
<210> 31
<211> 1773
<212> DNA
<213> Geobacillus
<400> 31
atggaacgca cctattattg tggtgaagtt ccggaaaccg cagttggtga acgtgttgtt 60
ctgaaaggtt gggttcagaa acgtcgtgat ttaggtggtc tgatttttat cgatctgcgt 120
gatcgtaccg gtattgttca ggttgttgca agtccggatg ttagcgcaga agcactggca 180
gcagcagaac gtgttcgtag cgaatatgtt ctgagcgttg aaggcaccgt tgttgcccgt 240
gcaccggaaa cagttaatcc gaatattgca accggtcgca ttgaaattca ggcagaacgt 300
attgaaatta tcaacgaagc aaaaacccct ccgtttagca ttagtgatga taccgatgca 360
gccgaagatg ttcgtctgaa atatcgttat ctggatctgc gtcgtccggt tatgtttcag 420
accctggcac tgcgtcataa aatcaccaaa accgttcgtg attttctgga tagcgaacgc 480
tttctggaaa ttgaaacccc gatgctgacc aaaagcacac cggaaggtgc acgtgattat 540
ctggttccga gccgtgttca tccgggtgaa ttttatgcac tgccgcagag tccgcagatc 600
tttaaacagc tgctgatggt tggtggtgtg gaacgttatt atcagattgc acgttgtttt 660
cgtgatgagg acctgcgtgc agatcgtcag ccggaattta cccagattga tattgaaatg 720
agcttcatcg agcaagagga tatcattgat ctgaccgaac gtatgatggc agcagttgtt 780
aaagcagcaa aaggtattga tattccgcgt ccgtttccgc gtattaccta tgatgaagca 840
atgagctgtt atggtagcga taaaccggat attcgttttg gtctggaact ggttgatgtg 900
agcgaaattg ttcgtgatag cgcatttcag gtttttgcgc gtgcagttaa agaaggtggt 960
caggttaaag caattaatgc aaaaggtgca gcaccgcgtt atagccgtaa agatattgat 1020
gcactgggcg aatttgcagg tcgttatggt gccaaaggtc tggcatggct gaaagcagaa 1080
ggtgaagaac tgaaaggtcc gattgcaaaa ttctttaccg atgaagaaca ggcagccctg 1140
cgtcgtgcac tggccgttga agatggtgac ctgctgctgt ttgttgcaga tgaaaaagca 1200
attgttgcag cagcactggg tgcgctgcgt ctgaaactgg gtaaagaact gggtctgatt 1260
gatgaagcca aactggcatt tctgtgggtt accgattggc ctctgctgga atacgatgaa 1320
gaggaaggtc gctattacgc agcacatcat ccgtttacca tgccggtgcg tgatgatatc 1380
ccgctgctgg aaaccaatcc gagcgcagtt cgtgcacagg catatgatct ggttctgaat 1440
ggttatgaat taggtggtgg tagcctgcgt atttttgaac gtgatgtgca agaaaaaatg 1500
tttcgtgccc tgggttttag cgaagaagaa gcacgtcgtc agtttggttt tctgttagaa 1560
gcatttgaat atggcacccc tccgcatggt ggtattgcac tgggtttaga tcgtctggtt 1620
atgctgctgg caggtcgtac caatctgcgc gataccattg catttccgaa aaccgccagc 1680
gcaagctgtc tgctgaccga agcaccgggt cctgttagcg acaaacagct ggaagaactg 1740
catctggcag ttgttctgcc ggaaaatgaa taa 1773
<210> 32
<211> 590
<212> PRT
<213> Geobacillus
<400> 32
Met Glu Arg Thr Tyr Tyr Cys Gly Glu Val Pro Glu Thr Ala Val Gly
1 5 10 15
Glu Arg Val Val Leu Lys Gly Trp Val Gln Lys Arg Arg Asp Leu Gly
20 25 30
Gly Leu Ile Phe Ile Asp Leu Arg Asp Arg Thr Gly Ile Val Gln Val
35 40 45
Val Ala Ser Pro Asp Val Ser Ala Glu Ala Leu Ala Ala Ala Glu Arg
50 55 60
Val Arg Ser Glu Tyr Val Leu Ser Val Glu Gly Thr Val Val Ala Arg
65 70 75 80
Ala Pro Glu Thr Val Asn Pro Asn Ile Ala Thr Gly Arg Ile Glu Ile
85 90 95
Gln Ala Glu Arg Ile Glu Ile Ile Asn Glu Ala Lys Thr Pro Pro Phe
100 105 110
Ser Ile Ser Asp Asp Thr Asp Ala Ala Glu Asp Val Arg Leu Lys Tyr
115 120 125
Arg Tyr Leu Asp Leu Arg Arg Pro Val Met Phe Gln Thr Leu Ala Leu
130 135 140
Arg His Lys Ile Thr Lys Thr Val Arg Asp Phe Leu Asp Ser Glu Arg
145 150 155 160
Phe Leu Glu Ile Glu Thr Pro Met Leu Thr Lys Ser Thr Pro Glu Gly
165 170 175
Ala Arg Asp Tyr Leu Val Pro Ser Arg Val His Pro Gly Glu Phe Tyr
180 185 190
Ala Leu Pro Gln Ser Pro Gln Ile Phe Lys Gln Leu Leu Met Val Gly
195 200 205
Gly Val Glu Arg Tyr Tyr Gln Ile Ala Arg Cys Phe Arg Asp Glu Asp
210 215 220
Leu Arg Ala Asp Arg Gln Pro Glu Phe Thr Gln Ile Asp Ile Glu Met
225 230 235 240
Ser Phe Ile Glu Gln Glu Asp Ile Ile Asp Leu Thr Glu Arg Met Met
245 250 255
Ala Ala Val Val Lys Ala Ala Lys Gly Ile Asp Ile Pro Arg Pro Phe
260 265 270
Pro Arg Ile Thr Tyr Asp Glu Ala Met Ser Cys Tyr Gly Ser Asp Lys
275 280 285
Pro Asp Ile Arg Phe Gly Leu Glu Leu Val Asp Val Ser Glu Ile Val
290 295 300
Arg Asp Ser Ala Phe Gln Val Phe Ala Arg Ala Val Lys Glu Gly Gly
305 310 315 320
Gln Val Lys Ala Ile Asn Ala Lys Gly Ala Ala Pro Arg Tyr Ser Arg
325 330 335
Lys Asp Ile Asp Ala Leu Gly Glu Phe Ala Gly Arg Tyr Gly Ala Lys
340 345 350
Gly Leu Ala Trp Leu Lys Ala Glu Gly Glu Glu Leu Lys Gly Pro Ile
355 360 365
Ala Lys Phe Phe Thr Asp Glu Glu Gln Ala Ala Leu Arg Arg Ala Leu
370 375 380
Ala Val Glu Asp Gly Asp Leu Leu Leu Phe Val Ala Asp Glu Lys Ala
385 390 395 400
Ile Val Ala Ala Ala Leu Gly Ala Leu Arg Leu Lys Leu Gly Lys Glu
405 410 415
Leu Gly Leu Ile Asp Glu Ala Lys Leu Ala Phe Leu Trp Val Thr Asp
420 425 430
Trp Pro Leu Leu Glu Tyr Asp Glu Glu Glu Gly Arg Tyr Tyr Ala Ala
435 440 445
His His Pro Phe Thr Met Pro Val Arg Asp Asp Ile Pro Leu Leu Glu
450 455 460
Thr Asn Pro Ser Ala Val Arg Ala Gln Ala Tyr Asp Leu Val Leu Asn
465 470 475 480
Gly Tyr Glu Leu Gly Gly Gly Ser Leu Arg Ile Phe Glu Arg Asp Val
485 490 495
Gln Glu Lys Met Phe Arg Ala Leu Gly Phe Ser Glu Glu Glu Ala Arg
500 505 510
Arg Gln Phe Gly Phe Leu Leu Glu Ala Phe Glu Tyr Gly Thr Pro Pro
515 520 525
His Gly Gly Ile Ala Leu Gly Leu Asp Arg Leu Val Met Leu Leu Ala
530 535 540
Gly Arg Thr Asn Leu Arg Asp Thr Ile Ala Phe Pro Lys Thr Ala Ser
545 550 555 560
Ala Ser Cys Leu Leu Thr Glu Ala Pro Gly Pro Val Ser Asp Lys Gln
565 570 575
Leu Glu Glu Leu His Leu Ala Val Val Leu Pro Glu Asn Glu
580 585 590
<210> 33
<211> 1401
<212> DNA
<213> Geobacillus
<400> 33
atgagcagca ttcgtctgta taataccctg acgcgtaaaa aagaaccgtt tgaaccgctg 60
gaaccgaaca aagttaaaat gtatgtttgt ggtccgaccg tgtataacta tattcatatt 120
ggtaatgccc gtgcagccat tgtgtttgat accattcgtc gttatctgga atttcgcggt 180
tatgatgtta cctatgtgag caattttacc gacgtggatg acaaactgat taaagcagca 240
cgtgaactgg gtgaaagcgt tccggcaatt gcagaacgtt ttattgaagc ctatttcgaa 300
gatattcagg ccctgggttg taaaaaagca gatattcatc cgcgtgtgac cgaaaatatc 360
gataccatta ttgaatttat ccaggcgctg atcgataaag gctatgcata tgaagttgat 420
ggcgacgttt attatcgtac ccgtaaattt cgcgaatatg gcaaactgag ccatcagagc 480
attgatgaac tgcaggcagg cgcacgtatt gaaattggtg aaaaaaaaga tgatccgctg 540
gattttgcac tgtggaaagc agcaaaagaa ggtgaaattt gttgggatag cccgtggggt 600
aaaggtcgtc ctggttggca tattgaatgt agcgcaatgg cacgtaaata tctgggtgat 660
acgattgata ttcatgccgg tggtcaggat ctgacctttc cgcatcatga aaatgaaatt 720
gcacagagcg aagcactgac cggtaaaccg tttgccaaat attggctgca taatggctat 780
ctgaacatca acaacgagaa aatgagcaaa agcctgggta attttgttct ggtgcatgat 840
attattcgcg agattgatcc gcaggttctg cgctttttta tgctgagcgt tcattatcgt 900
catccgatca attatagcga agaactgctg gaaagcgcac gtcgtggtct ggaacgtctg 960
aaaaccgcat atagcaatct gcagcaccgt ctgcaggcaa gcaccaatct gaccgataat 1020
gatgaagaat gggttagccg tattgccgat attcgtgcaa gctttattcg tgaaatggat 1080
gatgatttta acaccgccaa tggtattgcc gttctgtttg aactggcaaa acaggcaaat 1140
ctgtatctgc aagaaaaaac cacctccgaa aaagtgattc atgcatttct gcgtgaattt 1200
gaacagctgg cagatgttct gggtctgacc ctgaaacagg atgagctgct ggatgaagaa 1260
attgaagccc tgattcagaa acgtaatgaa gcccgtaaaa atcgtgattt tgccctggca 1320
gatcgtattc gtgatgaatt acgtgcgaaa aacatcatcc tggaagatac accgcagggc 1380
acccgttgga aacgtggtta a 1401
<210> 34
<211> 466
<212> PRT
<213> Geobacillus
<400> 34
Met Ser Ser Ile Arg Leu Tyr Asn Thr Leu Thr Arg Lys Lys Glu Pro
1 5 10 15
Phe Glu Pro Leu Glu Pro Asn Lys Val Lys Met Tyr Val Cys Gly Pro
20 25 30
Thr Val Tyr Asn Tyr Ile His Ile Gly Asn Ala Arg Ala Ala Ile Val
35 40 45
Phe Asp Thr Ile Arg Arg Tyr Leu Glu Phe Arg Gly Tyr Asp Val Thr
50 55 60
Tyr Val Ser Asn Phe Thr Asp Val Asp Asp Lys Leu Ile Lys Ala Ala
65 70 75 80
Arg Glu Leu Gly Glu Ser Val Pro Ala Ile Ala Glu Arg Phe Ile Glu
85 90 95
Ala Tyr Phe Glu Asp Ile Gln Ala Leu Gly Cys Lys Lys Ala Asp Ile
100 105 110
His Pro Arg Val Thr Glu Asn Ile Asp Thr Ile Ile Glu Phe Ile Gln
115 120 125
Ala Leu Ile Asp Lys Gly Tyr Ala Tyr Glu Val Asp Gly Asp Val Tyr
130 135 140
Tyr Arg Thr Arg Lys Phe Arg Glu Tyr Gly Lys Leu Ser His Gln Ser
145 150 155 160
Ile Asp Glu Leu Gln Ala Gly Ala Arg Ile Glu Ile Gly Glu Lys Lys
165 170 175
Asp Asp Pro Leu Asp Phe Ala Leu Trp Lys Ala Ala Lys Glu Gly Glu
180 185 190
Ile Cys Trp Asp Ser Pro Trp Gly Lys Gly Arg Pro Gly Trp His Ile
195 200 205
Glu Cys Ser Ala Met Ala Arg Lys Tyr Leu Gly Asp Thr Ile Asp Ile
210 215 220
His Ala Gly Gly Gln Asp Leu Thr Phe Pro His His Glu Asn Glu Ile
225 230 235 240
Ala Gln Ser Glu Ala Leu Thr Gly Lys Pro Phe Ala Lys Tyr Trp Leu
245 250 255
His Asn Gly Tyr Leu Asn Ile Asn Asn Glu Lys Met Ser Lys Ser Leu
260 265 270
Gly Asn Phe Val Leu Val His Asp Ile Ile Arg Glu Ile Asp Pro Gln
275 280 285
Val Leu Arg Phe Phe Met Leu Ser Val His Tyr Arg His Pro Ile Asn
290 295 300
Tyr Ser Glu Glu Leu Leu Glu Ser Ala Arg Arg Gly Leu Glu Arg Leu
305 310 315 320
Lys Thr Ala Tyr Ser Asn Leu Gln His Arg Leu Gln Ala Ser Thr Asn
325 330 335
Leu Thr Asp Asn Asp Glu Glu Trp Val Ser Arg Ile Ala Asp Ile Arg
340 345 350
Ala Ser Phe Ile Arg Glu Met Asp Asp Asp Phe Asn Thr Ala Asn Gly
355 360 365
Ile Ala Val Leu Phe Glu Leu Ala Lys Gln Ala Asn Leu Tyr Leu Gln
370 375 380
Glu Lys Thr Thr Ser Glu Lys Val Ile His Ala Phe Leu Arg Glu Phe
385 390 395 400
Glu Gln Leu Ala Asp Val Leu Gly Leu Thr Leu Lys Gln Asp Glu Leu
405 410 415
Leu Asp Glu Glu Ile Glu Ala Leu Ile Gln Lys Arg Asn Glu Ala Arg
420 425 430
Lys Asn Arg Asp Phe Ala Leu Ala Asp Arg Ile Arg Asp Glu Leu Arg
435 440 445
Ala Lys Asn Ile Ile Leu Glu Asp Thr Pro Gln Gly Thr Arg Trp Lys
450 455 460
Arg Gly
465
<210> 35
<211> 1665
<212> DNA
<213> E. coli
<400> 35
atgagcgaag cagaagcacg tccgaccaac tttattcgtc agattattga tgaagatctg 60
gccagcggta aacataccac cgttcatacc cgttttccgc ctgaaccgaa tggttatctg 120
catattggtc atgccaaaag catttgcctg aattttggta ttgcccagga ttataaaggt 180
cagtgcaatc tgcgtttcga tgataccaat ccggtgaaag aagatatcga atacgtcgag 240
agcatcaaaa atgatgttga atggctgggt tttcattgga gcggtaatgt tcgttatagc 300
agcgattatt ttgatcagct gcatgcctat gcaatcgaac tgattaacaa aggtctggcc 360
tatgttgatg aactgacacc ggaacaaatt cgtgaatatc gtggtacact gacccagcct 420
ggtaaaaata gcccgtatcg tgatcgtagc gttgaagaaa atctggccct gtttgaaaaa 480
atgcgtgccg gtggttttga agaaggtaaa gcctgtctgc gtgcaaaaat tgatatggca 540
agcccgttta ttgttatgcg tgatccggtt ctgtatcgca tcaaatttgc agaacatcat 600
cagaccggta acaaatggtg tatctatccg atgtatgatt tcacccattg cattagtgat 660
gccctggaag gtattaccca tagcctgtgt accctggaat ttcaggataa tcgtcgtctg 720
tatgattggg tgttagacaa tatcaccatt ccggtgcatc cgcgtcagta tgaatttagc 780
cgtctgaatc tggaatacac cgttatgagc aaacgtaaac tgaatctgct ggtgaccgat 840
aaacatgttg aaggttggga tgatccgcgt atgccgacca ttagcggtct gcgtcgtcgt 900
ggttataccg cagcaagcat ccgtgaattt tgtaaacgta ttggtgtgac caaacaggat 960
aacaccattg aaatggccag cctggaaagc tgtattcgcg aagatctgaa tgaaaatgca 1020
ccgcgtgcaa tggcagttat cgatccggtt aaactggtga tcgaaaatta tcaaggtgaa 1080
ggtgaaatgg tgaccatgcc gaatcatccg aataaaccgg aaatgggtag ccgtcaggtt 1140
ccgtttagcg gtgaaatttg gattgatcgt gcagattttc gtgaagaagc caacaaacag 1200
tataaacgtc tggttctggg taaagaagtt cgtctgcgta acgcctatgt tattaaagca 1260
gaacgtgttg aaaaagatgc cgaaggcaat attaccacca ttttttgtac ctatgacgca 1320
gataccctga gcaaagatcc ggcagatggt cgtaaagtta aaggtgttat tcattgggtt 1380
agcgcagcac atgcactgcc ggttgaaatt cgcctgtatg atcgtctgtt tagcgttccg 1440
aatccgggtg cagcagatga ttttctgagc gttattaatc cggaaagcct ggttattaaa 1500
cagggttttg ccgaaccgag cctgaaagat gcagttgcag gtaaagcatt tcagtttgaa 1560
cgcgaaggtt atttttgtct ggatagccgt catagcaccg cagaaaaacc ggtgtttaat 1620
cgtaccgttg gtctgcgtga tacctgggca aaagttggtg aataa 1665
<210> 36
<211> 554
<212> PRT
<213> E. coli
<400> 36
Met Ser Glu Ala Glu Ala Arg Pro Thr Asn Phe Ile Arg Gln Ile Ile
1 5 10 15
Asp Glu Asp Leu Ala Ser Gly Lys His Thr Thr Val His Thr Arg Phe
20 25 30
Pro Pro Glu Pro Asn Gly Tyr Leu His Ile Gly His Ala Lys Ser Ile
35 40 45
Cys Leu Asn Phe Gly Ile Ala Gln Asp Tyr Lys Gly Gln Cys Asn Leu
50 55 60
Arg Phe Asp Asp Thr Asn Pro Val Lys Glu Asp Ile Glu Tyr Val Glu
65 70 75 80
Ser Ile Lys Asn Asp Val Glu Trp Leu Gly Phe His Trp Ser Gly Asn
85 90 95
Val Arg Tyr Ser Ser Asp Tyr Phe Asp Gln Leu His Ala Tyr Ala Ile
100 105 110
Glu Leu Ile Asn Lys Gly Leu Ala Tyr Val Asp Glu Leu Thr Pro Glu
115 120 125
Gln Ile Arg Glu Tyr Arg Gly Thr Leu Thr Gln Pro Gly Lys Asn Ser
130 135 140
Pro Tyr Arg Asp Arg Ser Val Glu Glu Asn Leu Ala Leu Phe Glu Lys
145 150 155 160
Met Arg Ala Gly Gly Phe Glu Glu Gly Lys Ala Cys Leu Arg Ala Lys
165 170 175
Ile Asp Met Ala Ser Pro Phe Ile Val Met Arg Asp Pro Val Leu Tyr
180 185 190
Arg Ile Lys Phe Ala Glu His His Gln Thr Gly Asn Lys Trp Cys Ile
195 200 205
Tyr Pro Met Tyr Asp Phe Thr His Cys Ile Ser Asp Ala Leu Glu Gly
210 215 220
Ile Thr His Ser Leu Cys Thr Leu Glu Phe Gln Asp Asn Arg Arg Leu
225 230 235 240
Tyr Asp Trp Val Leu Asp Asn Ile Thr Ile Pro Val His Pro Arg Gln
245 250 255
Tyr Glu Phe Ser Arg Leu Asn Leu Glu Tyr Thr Val Met Ser Lys Arg
260 265 270
Lys Leu Asn Leu Leu Val Thr Asp Lys His Val Glu Gly Trp Asp Asp
275 280 285
Pro Arg Met Pro Thr Ile Ser Gly Leu Arg Arg Arg Gly Tyr Thr Ala
290 295 300
Ala Ser Ile Arg Glu Phe Cys Lys Arg Ile Gly Val Thr Lys Gln Asp
305 310 315 320
Asn Thr Ile Glu Met Ala Ser Leu Glu Ser Cys Ile Arg Glu Asp Leu
325 330 335
Asn Glu Asn Ala Pro Arg Ala Met Ala Val Ile Asp Pro Val Lys Leu
340 345 350
Val Ile Glu Asn Tyr Gln Gly Glu Gly Glu Met Val Thr Met Pro Asn
355 360 365
His Pro Asn Lys Pro Glu Met Gly Ser Arg Gln Val Pro Phe Ser Gly
370 375 380
Glu Ile Trp Ile Asp Arg Ala Asp Phe Arg Glu Glu Ala Asn Lys Gln
385 390 395 400
Tyr Lys Arg Leu Val Leu Gly Lys Glu Val Arg Leu Arg Asn Ala Tyr
405 410 415
Val Ile Lys Ala Glu Arg Val Glu Lys Asp Ala Glu Gly Asn Ile Thr
420 425 430
Thr Ile Phe Cys Thr Tyr Asp Ala Asp Thr Leu Ser Lys Asp Pro Ala
435 440 445
Asp Gly Arg Lys Val Lys Gly Val Ile His Trp Val Ser Ala Ala His
450 455 460
Ala Leu Pro Val Glu Ile Arg Leu Tyr Asp Arg Leu Phe Ser Val Pro
465 470 475 480
Asn Pro Gly Ala Ala Asp Asp Phe Leu Ser Val Ile Asn Pro Glu Ser
485 490 495
Leu Val Ile Lys Gln Gly Phe Ala Glu Pro Ser Leu Lys Asp Ala Val
500 505 510
Ala Gly Lys Ala Phe Gln Phe Glu Arg Glu Gly Tyr Phe Cys Leu Asp
515 520 525
Ser Arg His Ser Thr Ala Glu Lys Pro Val Phe Asn Arg Thr Val Gly
530 535 540
Leu Arg Asp Thr Trp Ala Lys Val Gly Glu
545 550
<210> 37
<211> 1473
<212> DNA
<213> Geobacillus
<400> 37
atggccaaag aagttcgcgt tcgttacgca ccgagtccga ccggtcatct gcatattggt 60
ggtgcacgta ccgcactgtt taattacctg tttgcacgtc atcatggtgg caaaatgatt 120
gtgcgtattg aagataccga tatcgaacgt aatgttgaag gtggtgaaaa aagccagctg 180
gaaaatctga aatggctggg cattgattat gatgaaagca ttgatcagga tggtggttat 240
ggtccgtatc gtcagaccga acgtctggat atttatcgca aatatgtgaa cgaactgctg 300
gaacagggtc atgcctataa atgtttttgt acaccggaag aactggaacg tgaacgtgaa 360
gcacagcgtg cagcaggtat tgcagcaccg cagtatagcg gtaaatgtcg tcatctgaca 420
ccggaacagg ttgccgaact ggaagcacag ggtaaaccgt ataccattcg tctgaaagtt 480
ccggaaggta aaacctatga attctatgat ctggtgcgtg gcaaagttgt gtttgaaagc 540
aaagatgttg gtggcgattg ggttattgtt aaagcaaatg gtattccgac ctataacttt 600
gccgttgtga ttgatgatca cctgatggaa atttcacatg tgtttcgtgg tgaagaacat 660
ctgagcaata ccccgaaaca gctgatggtg tatgaatatt ttggttggga accgcctcag 720
tttgcacatc tgaccctgat tgttaatgaa cagcgtaaaa aactgagcaa acgcgacgaa 780
agcattattc agtttgtgag ccagtataaa gaactgggtt atctgccgga agccatgttt 840
aacttttttg cactgttagg ttggtcaccg gaaggtgaag aagaaatctt taccaaagat 900
gaactgatcc gcatgtttga tgttagccgt ctgagcaaaa gcccgagtat gtttgatacc 960
aaaaagctga cctggatgaa caaccagtac atcaaaaaac tggatctgga tcgtctggtt 1020
gaactggcac tgccgcatct ggttaaagca ggtcgtctgc ctgcagatat gaccgatgag 1080
cagcgtcagt gggcacgtga tctgattgca ctgtatcaag agcagatgag ctatggtgca 1140
gaaattgttc cgctgagcga actgtttttc aaagaagaga ttgattacga ggatgaagca 1200
cgtcaggttc tggcagaaga acaggttccg gcagttctga gcacctttct ggaaagcgtt 1260
cgtgagctgg aaccgtttac cgcagatgaa attaaagcag caattaaagc cgttcagaaa 1320
gcaaccggtc agaaagggaa aaaactgttt atgccgattc gtgcagccgt tacaggtcag 1380
acccatggtc cggaactgcc gtttgcaatt cagctgctgg gtaaagaaaa agtgattgaa 1440
cgcctggaac gcgcactgca agaaaaattc taa 1473
<210> 38
<211> 490
<212> PRT
<213> Geobacillus
<400> 38
Met Ala Lys Glu Val Arg Val Arg Tyr Ala Pro Ser Pro Thr Gly His
1 5 10 15
Leu His Ile Gly Gly Ala Arg Thr Ala Leu Phe Asn Tyr Leu Phe Ala
20 25 30
Arg His His Gly Gly Lys Met Ile Val Arg Ile Glu Asp Thr Asp Ile
35 40 45
Glu Arg Asn Val Glu Gly Gly Glu Lys Ser Gln Leu Glu Asn Leu Lys
50 55 60
Trp Leu Gly Ile Asp Tyr Asp Glu Ser Ile Asp Gln Asp Gly Gly Tyr
65 70 75 80
Gly Pro Tyr Arg Gln Thr Glu Arg Leu Asp Ile Tyr Arg Lys Tyr Val
85 90 95
Asn Glu Leu Leu Glu Gln Gly His Ala Tyr Lys Cys Phe Cys Thr Pro
100 105 110
Glu Glu Leu Glu Arg Glu Arg Glu Ala Gln Arg Ala Ala Gly Ile Ala
115 120 125
Ala Pro Gln Tyr Ser Gly Lys Cys Arg His Leu Thr Pro Glu Gln Val
130 135 140
Ala Glu Leu Glu Ala Gln Gly Lys Pro Tyr Thr Ile Arg Leu Lys Val
145 150 155 160
Pro Glu Gly Lys Thr Tyr Glu Phe Tyr Asp Leu Val Arg Gly Lys Val
165 170 175
Val Phe Glu Ser Lys Asp Val Gly Gly Asp Trp Val Ile Val Lys Ala
180 185 190
Asn Gly Ile Pro Thr Tyr Asn Phe Ala Val Val Ile Asp Asp His Leu
195 200 205
Met Glu Ile Ser His Val Phe Arg Gly Glu Glu His Leu Ser Asn Thr
210 215 220
Pro Lys Gln Leu Met Val Tyr Glu Tyr Phe Gly Trp Glu Pro Pro Gln
225 230 235 240
Phe Ala His Leu Thr Leu Ile Val Asn Glu Gln Arg Lys Lys Leu Ser
245 250 255
Lys Arg Asp Glu Ser Ile Ile Gln Phe Val Ser Gln Tyr Lys Glu Leu
260 265 270
Gly Tyr Leu Pro Glu Ala Met Phe Asn Phe Phe Ala Leu Leu Gly Trp
275 280 285
Ser Pro Glu Gly Glu Glu Glu Ile Phe Thr Lys Asp Glu Leu Ile Arg
290 295 300
Met Phe Asp Val Ser Arg Leu Ser Lys Ser Pro Ser Met Phe Asp Thr
305 310 315 320
Lys Lys Leu Thr Trp Met Asn Asn Gln Tyr Ile Lys Lys Leu Asp Leu
325 330 335
Asp Arg Leu Val Glu Leu Ala Leu Pro His Leu Val Lys Ala Gly Arg
340 345 350
Leu Pro Ala Asp Met Thr Asp Glu Gln Arg Gln Trp Ala Arg Asp Leu
355 360 365
Ile Ala Leu Tyr Gln Glu Gln Met Ser Tyr Gly Ala Glu Ile Val Pro
370 375 380
Leu Ser Glu Leu Phe Phe Lys Glu Glu Ile Asp Tyr Glu Asp Glu Ala
385 390 395 400
Arg Gln Val Leu Ala Glu Glu Gln Val Pro Ala Val Leu Ser Thr Phe
405 410 415
Leu Glu Ser Val Arg Glu Leu Glu Pro Phe Thr Ala Asp Glu Ile Lys
420 425 430
Ala Ala Ile Lys Ala Val Gln Lys Ala Thr Gly Gln Lys Gly Lys Lys
435 440 445
Leu Phe Met Pro Ile Arg Ala Ala Val Thr Gly Gln Thr His Gly Pro
450 455 460
Glu Leu Pro Phe Ala Ile Gln Leu Leu Gly Lys Glu Lys Val Ile Glu
465 470 475 480
Arg Leu Glu Arg Ala Leu Gln Glu Lys Phe
485 490
<210> 39
<211> 1383
<212> DNA
<213> Geobacillus
<400> 39
atggcagtta ccatggaaga aattgttgca catgcaaaac atcgtggttt tgtttttccg 60
ggtagcgaaa tttatggtgg tctggcaaat acctgggatt atggtccgct gggtgttgaa 120
ctgaaaaata acattaaacg tgcctggtgg aaaaaattcg ttcaagaaag cccgtataat 180
gttggtctgg atgcagcaat tctgatgaat ccgcgtacct gggaagcaag cggtcatctg 240
ggtaacttta atgatccgat ggttgattgc aaacagtgta aagcacgtca tcgtgcagat 300
aaactgattg aaaaagccct ggaagaaaaa ggcattgaga tgattgttga tggtctgccg 360
ctggcaaaaa tggatgaact gattaaagaa tatgatatcg cctgtccgga atgtggtagc 420
cgtgatttta ccaatgttcg tcagtttaac ctgatgttca aaacctatca gggtgttacc 480
gaaagcagcg ccaatgaaat ttatctgcgt ccggaaaccg cacagggtat ttttgttaat 540
ttcaaaaatg tgcagcgcac catgcgtaaa aaactgccgt ttggtattgc acagattggc 600
aaaagctttc gcaacgaaat tacccctggt aattttacct ttcgcacccg tgaatttgag 660
cagatggaac tggaattttt ctgtaaaccg ggtgaagaac tgcagtggct ggaatattgg 720
aaacagtttt gtaaagaatg gctgctgagc ctgggtatga aagaagataa tattcgtctg 780
cgtgatcatg ccaaagaaga actgagccat tatagcaatg caaccaccga tatcgaatat 840
cattttccgt ttggttgggg tgaactgtgg ggtattgcaa gccgtaccga ttatgatctg 900
aaacgccata tggaatatag cggtgaagat ttccattacc tggatcaaga aaccaacgaa 960
cgttatattc cgtattgtat tgaaccgagt ctgggtgcag atcgtgttac cctggcattt 1020
atgattgatg cctatgatga agaggaactt gaagatggta caacccgtac cgtgatgcat 1080
ctgcatccgg cactggcacc gtataaagca gcagtgctgc cgttaagcaa aaaactggca 1140
gatggtgcac gtcgtattta tgaggaactg gcaaaacact tcatggtgga ttatgatgaa 1200
accggtagta ttggtaaacg ttatcgtcgt caggatgaaa ttggcacccc gttttgtatt 1260
acctatgatt ttgaaagcga acaggatggt caggttaccg ttcgtgatcg tgatacaatg 1320
gaacaggttc gtctgccgat tggcgaactg aaagcatttc tggaagagaa aatcgccttc 1380
taa 1383
<210> 40
<211> 460
<212> PRT
<213> Geobacillus
<400> 40
Met Ala Val Thr Met Glu Glu Ile Val Ala His Ala Lys His Arg Gly
1 5 10 15
Phe Val Phe Pro Gly Ser Glu Ile Tyr Gly Gly Leu Ala Asn Thr Trp
20 25 30
Asp Tyr Gly Pro Leu Gly Val Glu Leu Lys Asn Asn Ile Lys Arg Ala
35 40 45
Trp Trp Lys Lys Phe Val Gln Glu Ser Pro Tyr Asn Val Gly Leu Asp
50 55 60
Ala Ala Ile Leu Met Asn Pro Arg Thr Trp Glu Ala Ser Gly His Leu
65 70 75 80
Gly Asn Phe Asn Asp Pro Met Val Asp Cys Lys Gln Cys Lys Ala Arg
85 90 95
His Arg Ala Asp Lys Leu Ile Glu Lys Ala Leu Glu Glu Lys Gly Ile
100 105 110
Glu Met Ile Val Asp Gly Leu Pro Leu Ala Lys Met Asp Glu Leu Ile
115 120 125
Lys Glu Tyr Asp Ile Ala Cys Pro Glu Cys Gly Ser Arg Asp Phe Thr
130 135 140
Asn Val Arg Gln Phe Asn Leu Met Phe Lys Thr Tyr Gln Gly Val Thr
145 150 155 160
Glu Ser Ser Ala Asn Glu Ile Tyr Leu Arg Pro Glu Thr Ala Gln Gly
165 170 175
Ile Phe Val Asn Phe Lys Asn Val Gln Arg Thr Met Arg Lys Lys Leu
180 185 190
Pro Phe Gly Ile Ala Gln Ile Gly Lys Ser Phe Arg Asn Glu Ile Thr
195 200 205
Pro Gly Asn Phe Thr Phe Arg Thr Arg Glu Phe Glu Gln Met Glu Leu
210 215 220
Glu Phe Phe Cys Lys Pro Gly Glu Glu Leu Gln Trp Leu Glu Tyr Trp
225 230 235 240
Lys Gln Phe Cys Lys Glu Trp Leu Leu Ser Leu Gly Met Lys Glu Asp
245 250 255
Asn Ile Arg Leu Arg Asp His Ala Lys Glu Glu Leu Ser His Tyr Ser
260 265 270
Asn Ala Thr Thr Asp Ile Glu Tyr His Phe Pro Phe Gly Trp Gly Glu
275 280 285
Leu Trp Gly Ile Ala Ser Arg Thr Asp Tyr Asp Leu Lys Arg His Met
290 295 300
Glu Tyr Ser Gly Glu Asp Phe His Tyr Leu Asp Gln Glu Thr Asn Glu
305 310 315 320
Arg Tyr Ile Pro Tyr Cys Ile Glu Pro Ser Leu Gly Ala Asp Arg Val
325 330 335
Thr Leu Ala Phe Met Ile Asp Ala Tyr Asp Glu Glu Glu Leu Glu Asp
340 345 350
Gly Thr Thr Arg Thr Val Met His Leu His Pro Ala Leu Ala Pro Tyr
355 360 365
Lys Ala Ala Val Leu Pro Leu Ser Lys Lys Leu Ala Asp Gly Ala Arg
370 375 380
Arg Ile Tyr Glu Glu Leu Ala Lys His Phe Met Val Asp Tyr Asp Glu
385 390 395 400
Thr Gly Ser Ile Gly Lys Arg Tyr Arg Arg Gln Asp Glu Ile Gly Thr
405 410 415
Pro Phe Cys Ile Thr Tyr Asp Phe Glu Ser Glu Gln Asp Gly Gln Val
420 425 430
Thr Val Arg Asp Arg Asp Thr Met Glu Gln Val Arg Leu Pro Ile Gly
435 440 445
Glu Leu Lys Ala Phe Leu Glu Glu Lys Ile Ala Phe
450 455 460
<210> 41
<211> 1281
<212> DNA
<213> Geobacillus
<400> 41
atggcatttc agattccgcg tggcacccag gatgttctgc ctggtgatac cgaaaaatgg 60
cagtatgttg aacatgttgc acgtaatctg tgtagccgtt atggttatcg tgaaattcgt 120
accccgattt ttgaacacac cgaactgttt ctgcgtggtg tgggtgatac caccgatatt 180
gttcagaaag aaatgtatac cttcgaggat aaaggtggtc gtgcactgac cctgcgtccg 240
gaaggcaccg caccggttgt tcgtgcattt gtggaacata aactgtatgg tagtccgcat 300
cagccgctga aactgtatta ttcaggtccg atgtttcgtt atgaacgtcc tgaagcaggt 360
cgttttcgtc agtttgttca gtttggtgtt gaagcactgg gtagcagcga tccggcaatt 420
gatgcagaag ttatggcact ggcaatgcat atttatgaag ccctgggtct gaaacgtatt 480
cgtctggtga ttaatagcct gggtgatctg gatagccgtc gtgcacatcg tgaagcgctg 540
gttcgtcatt ttagcagccg tattcatgaa ctgtgtccgg attgtcagac ccgtctgcat 600
accaatccgc tgcgtattct ggattgtaaa aaagatcgtg atcatgagct gatggcaacc 660
gcaccgagca tcctggatta tctgaatgaa gatagccgtg cctatttcga gaaagtgaaa 720
cagtatctga ccaatctggg tattccgttt gttattgata gtcgtctggt tcgtggtctg 780
gattattaca atcataccac ctttgaaatc atgagcgaag ccgaaggttt tggtgcagca 840
gcaaccctgt gtggtggtgg tcgttataat ggtctggttc aagaaattgg tggtccggaa 900
acacctggta ttggttttgc actgagcatt gaacgtctgc tggcagcact ggatgccgaa 960
ggtgttgaac tgccggttga aagtggcctg gattgttatg ttgttgcagt tggtgaacgt 1020
gcaaaagatg aagcagtgcg tctggtttat gccctgcgtc gtagcggtct gcgtgttgat 1080
caggattacc tgggtcgtaa actgaaagca cagctgaaag cagcagatcg tctgggtgca 1140
agctttgttg caattattgg tgatgaggaa ctggaacgtc aagaagcagc agttaaacat 1200
atggcaagcg gtgaacagac caatgttccg ctgggtgaac tggcacattt tctgcatgaa 1260
cgtattggca aagaagaata a 1281
<210> 42
<211> 426
<212> PRT
<213> Geobacillus
<400> 42
Met Ala Phe Gln Ile Pro Arg Gly Thr Gln Asp Val Leu Pro Gly Asp
1 5 10 15
Thr Glu Lys Trp Gln Tyr Val Glu His Val Ala Arg Asn Leu Cys Ser
20 25 30
Arg Tyr Gly Tyr Arg Glu Ile Arg Thr Pro Ile Phe Glu His Thr Glu
35 40 45
Leu Phe Leu Arg Gly Val Gly Asp Thr Thr Asp Ile Val Gln Lys Glu
50 55 60
Met Tyr Thr Phe Glu Asp Lys Gly Gly Arg Ala Leu Thr Leu Arg Pro
65 70 75 80
Glu Gly Thr Ala Pro Val Val Arg Ala Phe Val Glu His Lys Leu Tyr
85 90 95
Gly Ser Pro His Gln Pro Leu Lys Leu Tyr Tyr Ser Gly Pro Met Phe
100 105 110
Arg Tyr Glu Arg Pro Glu Ala Gly Arg Phe Arg Gln Phe Val Gln Phe
115 120 125
Gly Val Glu Ala Leu Gly Ser Ser Asp Pro Ala Ile Asp Ala Glu Val
130 135 140
Met Ala Leu Ala Met His Ile Tyr Glu Ala Leu Gly Leu Lys Arg Ile
145 150 155 160
Arg Leu Val Ile Asn Ser Leu Gly Asp Leu Asp Ser Arg Arg Ala His
165 170 175
Arg Glu Ala Leu Val Arg His Phe Ser Ser Arg Ile His Glu Leu Cys
180 185 190
Pro Asp Cys Gln Thr Arg Leu His Thr Asn Pro Leu Arg Ile Leu Asp
195 200 205
Cys Lys Lys Asp Arg Asp His Glu Leu Met Ala Thr Ala Pro Ser Ile
210 215 220
Leu Asp Tyr Leu Asn Glu Asp Ser Arg Ala Tyr Phe Glu Lys Val Lys
225 230 235 240
Gln Tyr Leu Thr Asn Leu Gly Ile Pro Phe Val Ile Asp Ser Arg Leu
245 250 255
Val Arg Gly Leu Asp Tyr Tyr Asn His Thr Thr Phe Glu Ile Met Ser
260 265 270
Glu Ala Glu Gly Phe Gly Ala Ala Ala Thr Leu Cys Gly Gly Gly Arg
275 280 285
Tyr Asn Gly Leu Val Gln Glu Ile Gly Gly Pro Glu Thr Pro Gly Ile
290 295 300
Gly Phe Ala Leu Ser Ile Glu Arg Leu Leu Ala Ala Leu Asp Ala Glu
305 310 315 320
Gly Val Glu Leu Pro Val Glu Ser Gly Leu Asp Cys Tyr Val Val Ala
325 330 335
Val Gly Glu Arg Ala Lys Asp Glu Ala Val Arg Leu Val Tyr Ala Leu
340 345 350
Arg Arg Ser Gly Leu Arg Val Asp Gln Asp Tyr Leu Gly Arg Lys Leu
355 360 365
Lys Ala Gln Leu Lys Ala Ala Asp Arg Leu Gly Ala Ser Phe Val Ala
370 375 380
Ile Ile Gly Asp Glu Glu Leu Glu Arg Gln Glu Ala Ala Val Lys His
385 390 395 400
Met Ala Ser Gly Glu Gln Thr Asn Val Pro Leu Gly Glu Leu Ala His
405 410 415
Phe Leu His Glu Arg Ile Gly Lys Glu Glu
420 425
<210> 43
<211> 2775
<212> DNA
<213> Geobacillus stearothermophilus
<400> 43
atggactaca aagaaaccct gctgatgccg cagaccgaat ttccgatgcg tggtaatctg 60
ccgaaacgtg aaccggaaat gcagaaaaaa tgggaagaga tggatatcta ccgcaaagtt 120
caagaacgta ccaaaggtcg tccgctgttt gttctgcatg atggtccgcc ttatgcaaat 180
ggtgatattc atatgggtca tgccctgaac aaaatcctga aagatattat cgtgcgctat 240
aagagcatga atggttattg tgcaccgtat gttccaggtt gggataccca tggtctgccg 300
attgaaaccg cactggcaaa acagggtgtt gatcgtaaaa gcatgagcgt tgcagaattt 360
cgtaaacgtt gtgaacagta tgcctatgag cagattgata atcagcgtcg tcagtttaaa 420
cgtctgggtg ttcgtggtga ttgggataat ccgtatatta ccctgaaacc ggaatatgaa 480
gcacagcaga ttaaagtgtt tggcgagatg gcaaaaaaag gcctgatcta taaaggtctg 540
aaacctgttt attggagccc gagcagcgaa agtgcactgg cagaagcaga aattgagtat 600
aaagataaac gctccccgag catttatgtt gcctttccgg ttaaagatgg taaaggtgtt 660
ctggaaggtg atgaacgtat tgtgatttgg accaccacac cgtggaccat tccggcaaat 720
ctggcaattg cagttcatcc ggatctggat tatcatgttg ttgatgttag cggtaaacgt 780
tatgttgttg cagcagcact ggccgaaagc gttgcaaaag aaattggttg ggatgcatgg 840
tcagttgtga aaaccgttaa aggtaaagaa ctggaatatg tggttgcgaa acacccgttt 900
tatgaacgtg atagcctggt tgtttgtggt gaacatgtga ccaccgatgc aggcaccggt 960
tgtgttcata ccgcacctgg tcatggtgaa gatgattttc tggttggtca gaaatatggc 1020
ctgccggttc tgtgtccggt ggatgaacgt ggttatatga ccgaagaagc accgggtttt 1080
gaaggtatgt tttatgagga tgccaacaaa gcgattacgc agaaactgga agaagttggc 1140
gcactgctga aactgggttt tattacccat agctatccgc atgattggcg taccaaacag 1200
ccgaccattt ttcgtgcaac cacacagtgg tttgcaagca ttgataaaat tcgcaatgaa 1260
ctgctgcagg ccatcaaaga aacaaaatgg atcccggaat ggggtgaaat tcgcattcat 1320
aacatggttc gtgatcgcgg tgattggtgt attagccgtc agcgtgcatg gggtgttccg 1380
attccggtgt tttatggtga aaatggtgaa ccgattatca ccgatgaaac cattgaacat 1440
gttagcaacc tgtttcgtca gtatggtagc aatgtttggt ttgaacgtga agcaaaagat 1500
ctgctgccgg aaggttttac ccatccgagc agcccgaatg gtatttttac aaaagaaacc 1560
gatatcatgg acgtgtggtt tgatagcggt agcagccatc aggcagttct ggtggaacgt 1620
gatgatctga tgcgtccggc agatctgtat ctggaaggca gcgatcagta tcgtggttgg 1680
tttaatagca gcctgagcac cgcagttgca gtgaccggta aagcaccgta taaaggtgtg 1740
ctgagccatg gttttgtgct ggatggtgaa ggtcgtaaaa tgagcaaaag cctgggtaat 1800
gttgttgttc ctgcaaaagt tatggaacag tttggtgcag atattctgcg tctgtgggtt 1860
gccagcgttg attatcaggc agatgttcgt attagcgatc atattctgaa acaggtgagc 1920
gaagtgtatc gcaaaattcg taataccttt cgctttatgc tgggtaacct gtttgatttt 1980
gatccgaatc agaatgcagt tccgattggt gaactgggtg aagttgatcg ttatatgctg 2040
gccaaactga ataaactgat cgccaaagtg aaaaaagcct atgatagcta cgatttcgca 2100
gccgtttatc atgaaatgaa ccatttttgt accgttgaac tgagcgcctt ttatctggat 2160
atggcaaaag atatcctgta tatcgaagca gcagatagcc gtgcacgtcg tgcagttcag 2220
accgttctgt atgaaaccgt tgttgcactg gcgaaactga ttgcaccgat tctgccgcat 2280
accgcagatg aagtttggga acatattccg aatcgtcgtg aaaatgtgga aagcgttcag 2340
ctgaccgata tgccggaacc gattgcaatt gatggcgaag aggcactgct ggcaaaatgg 2400
gatgccttta tggatgttcg tgatgatatg ctgaaagcac tggaaaatgc ccgtaacgaa 2460
aaagtgattg gtaaaagcct gaccgcaagc gttattgttt atccgaaaga tgaagcacgt 2520
aaactgctgg cgagcctgga tgccgatctg cgtcagctgc tgattgttag cgcatttagc 2580
attgcagatg aaccgtatga tgctgcccct gcagaagccg aacgtctgga tcatgttgcc 2640
gttctggttc gtcctgccga aggtgaaacc tgcgaacgtt gttggaccgt tacaccggca 2700
gttggtcagg atccgagcca tccgaccttt tgtccgcgtt gtgcacatat tgttaacgaa 2760
cattatagcg cctaa 2775
<210> 44
<211> 924
<212> PRT
<213> Geobacillus stearothermophilus
<400> 44
Met Asp Tyr Lys Glu Thr Leu Leu Met Pro Gln Thr Glu Phe Pro Met
1 5 10 15
Arg Gly Asn Leu Pro Lys Arg Glu Pro Glu Met Gln Lys Lys Trp Glu
20 25 30
Glu Met Asp Ile Tyr Arg Lys Val Gln Glu Arg Thr Lys Gly Arg Pro
35 40 45
Leu Phe Val Leu His Asp Gly Pro Pro Tyr Ala Asn Gly Asp Ile His
50 55 60
Met Gly His Ala Leu Asn Lys Ile Leu Lys Asp Ile Ile Val Arg Tyr
65 70 75 80
Lys Ser Met Asn Gly Tyr Cys Ala Pro Tyr Val Pro Gly Trp Asp Thr
85 90 95
His Gly Leu Pro Ile Glu Thr Ala Leu Ala Lys Gln Gly Val Asp Arg
100 105 110
Lys Ser Met Ser Val Ala Glu Phe Arg Lys Arg Cys Glu Gln Tyr Ala
115 120 125
Tyr Glu Gln Ile Asp Asn Gln Arg Arg Gln Phe Lys Arg Leu Gly Val
130 135 140
Arg Gly Asp Trp Asp Asn Pro Tyr Ile Thr Leu Lys Pro Glu Tyr Glu
145 150 155 160
Ala Gln Gln Ile Lys Val Phe Gly Glu Met Ala Lys Lys Gly Leu Ile
165 170 175
Tyr Lys Gly Leu Lys Pro Val Tyr Trp Ser Pro Ser Ser Glu Ser Ala
180 185 190
Leu Ala Glu Ala Glu Ile Glu Tyr Lys Asp Lys Arg Ser Pro Ser Ile
195 200 205
Tyr Val Ala Phe Pro Val Lys Asp Gly Lys Gly Val Leu Glu Gly Asp
210 215 220
Glu Arg Ile Val Ile Trp Thr Thr Thr Pro Trp Thr Ile Pro Ala Asn
225 230 235 240
Leu Ala Ile Ala Val His Pro Asp Leu Asp Tyr His Val Val Asp Val
245 250 255
Ser Gly Lys Arg Tyr Val Val Ala Ala Ala Leu Ala Glu Ser Val Ala
260 265 270
Lys Glu Ile Gly Trp Asp Ala Trp Ser Val Val Lys Thr Val Lys Gly
275 280 285
Lys Glu Leu Glu Tyr Val Val Ala Lys His Pro Phe Tyr Glu Arg Asp
290 295 300
Ser Leu Val Val Cys Gly Glu His Val Thr Thr Asp Ala Gly Thr Gly
305 310 315 320
Cys Val His Thr Ala Pro Gly His Gly Glu Asp Asp Phe Leu Val Gly
325 330 335
Gln Lys Tyr Gly Leu Pro Val Leu Cys Pro Val Asp Glu Arg Gly Tyr
340 345 350
Met Thr Glu Glu Ala Pro Gly Phe Glu Gly Met Phe Tyr Glu Asp Ala
355 360 365
Asn Lys Ala Ile Thr Gln Lys Leu Glu Glu Val Gly Ala Leu Leu Lys
370 375 380
Leu Gly Phe Ile Thr His Ser Tyr Pro His Asp Trp Arg Thr Lys Gln
385 390 395 400
Pro Thr Ile Phe Arg Ala Thr Thr Gln Trp Phe Ala Ser Ile Asp Lys
405 410 415
Ile Arg Asn Glu Leu Leu Gln Ala Ile Lys Glu Thr Lys Trp Ile Pro
420 425 430
Glu Trp Gly Glu Ile Arg Ile His Asn Met Val Arg Asp Arg Gly Asp
435 440 445
Trp Cys Ile Ser Arg Gln Arg Ala Trp Gly Val Pro Ile Pro Val Phe
450 455 460
Tyr Gly Glu Asn Gly Glu Pro Ile Ile Thr Asp Glu Thr Ile Glu His
465 470 475 480
Val Ser Asn Leu Phe Arg Gln Tyr Gly Ser Asn Val Trp Phe Glu Arg
485 490 495
Glu Ala Lys Asp Leu Leu Pro Glu Gly Phe Thr His Pro Ser Ser Pro
500 505 510
Asn Gly Ile Phe Thr Lys Glu Thr Asp Ile Met Asp Val Trp Phe Asp
515 520 525
Ser Gly Ser Ser His Gln Ala Val Leu Val Glu Arg Asp Asp Leu Met
530 535 540
Arg Pro Ala Asp Leu Tyr Leu Glu Gly Ser Asp Gln Tyr Arg Gly Trp
545 550 555 560
Phe Asn Ser Ser Leu Ser Thr Ala Val Ala Val Thr Gly Lys Ala Pro
565 570 575
Tyr Lys Gly Val Leu Ser His Gly Phe Val Leu Asp Gly Glu Gly Arg
580 585 590
Lys Met Ser Lys Ser Leu Gly Asn Val Val Val Pro Ala Lys Val Met
595 600 605
Glu Gln Phe Gly Ala Asp Ile Leu Arg Leu Trp Val Ala Ser Val Asp
610 615 620
Tyr Gln Ala Asp Val Arg Ile Ser Asp His Ile Leu Lys Gln Val Ser
625 630 635 640
Glu Val Tyr Arg Lys Ile Arg Asn Thr Phe Arg Phe Met Leu Gly Asn
645 650 655
Leu Phe Asp Phe Asp Pro Asn Gln Asn Ala Val Pro Ile Gly Glu Leu
660 665 670
Gly Glu Val Asp Arg Tyr Met Leu Ala Lys Leu Asn Lys Leu Ile Ala
675 680 685
Lys Val Lys Lys Ala Tyr Asp Ser Tyr Asp Phe Ala Ala Val Tyr His
690 695 700
Glu Met Asn His Phe Cys Thr Val Glu Leu Ser Ala Phe Tyr Leu Asp
705 710 715 720
Met Ala Lys Asp Ile Leu Tyr Ile Glu Ala Ala Asp Ser Arg Ala Arg
725 730 735
Arg Ala Val Gln Thr Val Leu Tyr Glu Thr Val Val Ala Leu Ala Lys
740 745 750
Leu Ile Ala Pro Ile Leu Pro His Thr Ala Asp Glu Val Trp Glu His
755 760 765
Ile Pro Asn Arg Arg Glu Asn Val Glu Ser Val Gln Leu Thr Asp Met
770 775 780
Pro Glu Pro Ile Ala Ile Asp Gly Glu Glu Ala Leu Leu Ala Lys Trp
785 790 795 800
Asp Ala Phe Met Asp Val Arg Asp Asp Met Leu Lys Ala Leu Glu Asn
805 810 815
Ala Arg Asn Glu Lys Val Ile Gly Lys Ser Leu Thr Ala Ser Val Ile
820 825 830
Val Tyr Pro Lys Asp Glu Ala Arg Lys Leu Leu Ala Ser Leu Asp Ala
835 840 845
Asp Leu Arg Gln Leu Leu Ile Val Ser Ala Phe Ser Ile Ala Asp Glu
850 855 860
Pro Tyr Asp Ala Ala Pro Ala Glu Ala Glu Arg Leu Asp His Val Ala
865 870 875 880
Val Leu Val Arg Pro Ala Glu Gly Glu Thr Cys Glu Arg Cys Trp Thr
885 890 895
Val Thr Pro Ala Val Gly Gln Asp Pro Ser His Pro Thr Phe Cys Pro
900 905 910
Arg Cys Ala His Ile Val Asn Glu His Tyr Ser Ala
915 920
<210> 45
<211> 2418
<212> DNA
<213> Geobacillus stearothermophilus
<400> 45
atgagcttta accaccgtga aatcgaacag aaatggcagg attattggga gaagaataaa 60
acctttcgta caccggatga tgatgacaaa ccgaaattct atgtgctgga tatgtttccg 120
tatccgagcg gtgcaggtct gcatgttggt catccggaag gttataccgc aaccgatatt 180
ctggcacgta tgaaacgtat gcagggttat aatgttctgc atccgatggg ttgggatgca 240
tttggtctgc ctgcagaaca gtatgcactg gataccggta atgatccggc agaatttacc 300
cagaaaaaca tcgataactt tcgtcgccag attaaaagcc tgggttttag ctatgattgg 360
gatcgtgaaa tcaataccac cgatccgaat tattacaaat ggacccagtg gatcttcctg 420
aaactgtatg aaaaaggtct ggcctatatg gatgaagttc cggttaattg gtgtccggca 480
ctgggcaccg ttctggcaaa tgaagaagtt attaacggtc gtagcgaacg tggtggccat 540
ccggttattc gtaaaccgat gcgtcagtgg atgctgaaaa ttaccgcata tgcagatcgt 600
ctgctggaag atctggaaga attagattgg cctgaaagca tcaaagaaat gcagcgtaat 660
tggattggtc gtagtgaagg tgcagaaatt gaatttgcag ttgatggtca cgatgaaacc 720
tttaccgttt ttaccacacg tccggataca ctgtttggtg caacctatac cgtgctggca 780
ccggaacatc cgctggttga aaaaatcacc actccggaac agaaacctgc cgttgatgca 840
tatctgaaag aaattcagag caaaagcgat ctggaacgta ccgatctggc caaagaaaaa 900
accggtgtgt ttaccggtgc atatgccatt catcctgtta ccggtgatcg cctgccgatt 960
tggattgcag attatgttct gatgagctat ggtacaggtg caattatggc agttccggca 1020
catgatgaac gtgattatga attcgccaaa aaattccatc tgccgatgaa agaagttgtt 1080
gcaggcggta atattgagaa agaagcatat acaggcgacg gcgaacatat taacagcgaa 1140
tttctgaatg gcctgaataa acaagaggcc atcgataaaa tgattgcctg gctggaagaa 1200
catggtaaag gtcgtaaaaa agttagctat cgtctgcgtg attggctgtt tagccgtcag 1260
cgttattggg gtgaaccgat tccgattatt cattgggaag atggcaccat gacaccggtt 1320
ccggaagaag aactgccgct ggttctgccg aaaaccgatg aaattcgtcc gagcggcacc 1380
ggtgaaagtc cgctggcaaa tattgaagaa tgggttaatg ttgtggatcc gaaaacgggt 1440
aaaaaaggtc gtcgcgaaac caataccatg ccgcagtggg caggtagctg ttggtattat 1500
ctgcgttata ttgatccgca caacgataaa cagctggcag atccggaaaa actgaaaaaa 1560
tggctgccgg ttgatgtgta tattggtggt gccgaacatg cagtgctgca tctgctgtat 1620
gcacgttttt ggcataaatt tctgtatgac ctgggtattg ttccgaccaa agaaccgttt 1680
cagaaactgt ttaatcaggg tatgattctg ggcgagaaca acgaaaaaat gagcaaaagt 1740
aaaggcaatg tggtgaaccc ggatgatatt attgaaagcc atggtgcaga taccctgcgt 1800
ctgtatgaga tgtttatggg tccgctggaa gcaagcattg catggtcaac caaaggcctg 1860
gatggtgcac gtcgttttct ggatcgtgtt tggcgtctgt ttgttaccga aaatggtgaa 1920
ctgaatccga acattgttga tgaaccggca aatgataccc tggaacgcat ttatcatcag 1980
accgttaaaa aagtgaccga ggattatgaa gccctgcgtt ttaataccgc aattagccag 2040
ctgatggtgt ttattaacga agcctataaa gccgagcaga tgaaaaaaga atatatggaa 2100
ggcttcgtga aactgctgag tccggtttgt ccgcatattg gtgaagaact gtggcagaaa 2160
ctgggtcata ccgataccat tgcatatgaa ccgtggccga cctatgatga aaccaaactg 2220
gttgaagatg tggtggaaat tgttgtgcag attaatggta aagtgcgtag tcgcctgcat 2280
gtgcctgttg atctgcctaa agaagcctta gaagaacgcg cactggcgga tgaaaagatt 2340
aaagaacagc tggaaggtaa aaccgtgcgt aaagttattg ccgttccggg taaactggtt 2400
aatattgttg ccaactaa 2418
<210> 46
<211> 805
<212> PRT
<213> Geobacillus stearothermophilus
<400> 46
Met Ser Phe Asn His Arg Glu Ile Glu Gln Lys Trp Gln Asp Tyr Trp
1 5 10 15
Glu Lys Asn Lys Thr Phe Arg Thr Pro Asp Asp Asp Asp Lys Pro Lys
20 25 30
Phe Tyr Val Leu Asp Met Phe Pro Tyr Pro Ser Gly Ala Gly Leu His
35 40 45
Val Gly His Pro Glu Gly Tyr Thr Ala Thr Asp Ile Leu Ala Arg Met
50 55 60
Lys Arg Met Gln Gly Tyr Asn Val Leu His Pro Met Gly Trp Asp Ala
65 70 75 80
Phe Gly Leu Pro Ala Glu Gln Tyr Ala Leu Asp Thr Gly Asn Asp Pro
85 90 95
Ala Glu Phe Thr Gln Lys Asn Ile Asp Asn Phe Arg Arg Gln Ile Lys
100 105 110
Ser Leu Gly Phe Ser Tyr Asp Trp Asp Arg Glu Ile Asn Thr Thr Asp
115 120 125
Pro Asn Tyr Tyr Lys Trp Thr Gln Trp Ile Phe Leu Lys Leu Tyr Glu
130 135 140
Lys Gly Leu Ala Tyr Met Asp Glu Val Pro Val Asn Trp Cys Pro Ala
145 150 155 160
Leu Gly Thr Val Leu Ala Asn Glu Glu Val Ile Asn Gly Arg Ser Glu
165 170 175
Arg Gly Gly His Pro Val Ile Arg Lys Pro Met Arg Gln Trp Met Leu
180 185 190
Lys Ile Thr Ala Tyr Ala Asp Arg Leu Leu Glu Asp Leu Glu Glu Leu
195 200 205
Asp Trp Pro Glu Ser Ile Lys Glu Met Gln Arg Asn Trp Ile Gly Arg
210 215 220
Ser Glu Gly Ala Glu Ile Glu Phe Ala Val Asp Gly His Asp Glu Thr
225 230 235 240
Phe Thr Val Phe Thr Thr Arg Pro Asp Thr Leu Phe Gly Ala Thr Tyr
245 250 255
Thr Val Leu Ala Pro Glu His Pro Leu Val Glu Lys Ile Thr Thr Pro
260 265 270
Glu Gln Lys Pro Ala Val Asp Ala Tyr Leu Lys Glu Ile Gln Ser Lys
275 280 285
Ser Asp Leu Glu Arg Thr Asp Leu Ala Lys Glu Lys Thr Gly Val Phe
290 295 300
Thr Gly Ala Tyr Ala Ile His Pro Val Thr Gly Asp Arg Leu Pro Ile
305 310 315 320
Trp Ile Ala Asp Tyr Val Leu Met Ser Tyr Gly Thr Gly Ala Ile Met
325 330 335
Ala Val Pro Ala His Asp Glu Arg Asp Tyr Glu Phe Ala Lys Lys Phe
340 345 350
His Leu Pro Met Lys Glu Val Val Ala Gly Gly Asn Ile Glu Lys Glu
355 360 365
Ala Tyr Thr Gly Asp Gly Glu His Ile Asn Ser Glu Phe Leu Asn Gly
370 375 380
Leu Asn Lys Gln Glu Ala Ile Asp Lys Met Ile Ala Trp Leu Glu Glu
385 390 395 400
His Gly Lys Gly Arg Lys Lys Val Ser Tyr Arg Leu Arg Asp Trp Leu
405 410 415
Phe Ser Arg Gln Arg Tyr Trp Gly Glu Pro Ile Pro Ile Ile His Trp
420 425 430
Glu Asp Gly Thr Met Thr Pro Val Pro Glu Glu Glu Leu Pro Leu Val
435 440 445
Leu Pro Lys Thr Asp Glu Ile Arg Pro Ser Gly Thr Gly Glu Ser Pro
450 455 460
Leu Ala Asn Ile Glu Glu Trp Val Asn Val Val Asp Pro Lys Thr Gly
465 470 475 480
Lys Lys Gly Arg Arg Glu Thr Asn Thr Met Pro Gln Trp Ala Gly Ser
485 490 495
Cys Trp Tyr Tyr Leu Arg Tyr Ile Asp Pro His Asn Asp Lys Gln Leu
500 505 510
Ala Asp Pro Glu Lys Leu Lys Lys Trp Leu Pro Val Asp Val Tyr Ile
515 520 525
Gly Gly Ala Glu His Ala Val Leu His Leu Leu Tyr Ala Arg Phe Trp
530 535 540
His Lys Phe Leu Tyr Asp Leu Gly Ile Val Pro Thr Lys Glu Pro Phe
545 550 555 560
Gln Lys Leu Phe Asn Gln Gly Met Ile Leu Gly Glu Asn Asn Glu Lys
565 570 575
Met Ser Lys Ser Lys Gly Asn Val Val Asn Pro Asp Asp Ile Ile Glu
580 585 590
Ser His Gly Ala Asp Thr Leu Arg Leu Tyr Glu Met Phe Met Gly Pro
595 600 605
Leu Glu Ala Ser Ile Ala Trp Ser Thr Lys Gly Leu Asp Gly Ala Arg
610 615 620
Arg Phe Leu Asp Arg Val Trp Arg Leu Phe Val Thr Glu Asn Gly Glu
625 630 635 640
Leu Asn Pro Asn Ile Val Asp Glu Pro Ala Asn Asp Thr Leu Glu Arg
645 650 655
Ile Tyr His Gln Thr Val Lys Lys Val Thr Glu Asp Tyr Glu Ala Leu
660 665 670
Arg Phe Asn Thr Ala Ile Ser Gln Leu Met Val Phe Ile Asn Glu Ala
675 680 685
Tyr Lys Ala Glu Gln Met Lys Lys Glu Tyr Met Glu Gly Phe Val Lys
690 695 700
Leu Leu Ser Pro Val Cys Pro His Ile Gly Glu Glu Leu Trp Gln Lys
705 710 715 720
Leu Gly His Thr Asp Thr Ile Ala Tyr Glu Pro Trp Pro Thr Tyr Asp
725 730 735
Glu Thr Lys Leu Val Glu Asp Val Val Glu Ile Val Val Gln Ile Asn
740 745 750
Gly Lys Val Arg Ser Arg Leu His Val Pro Val Asp Leu Pro Lys Glu
755 760 765
Ala Leu Glu Glu Arg Ala Leu Ala Asp Glu Lys Ile Lys Glu Gln Leu
770 775 780
Glu Gly Lys Thr Val Arg Lys Val Ile Ala Val Pro Gly Lys Leu Val
785 790 795 800
Asn Ile Val Ala Asn
805
<210> 47
<211> 1485
<212> DNA
<213> Geobacillus stearothermophilus
<400> 47
atgagccatg aagaactgaa tgatcagctg cgtgttcgtc gtgaaaaact gaaaaaaatc 60
gaagaactgg gcgttgatcc gtttggtaaa cgttttgaac gtacccataa agcccaagaa 120
ctgtttgaac tgtatggtga tctgagcaaa gaggaactgg aagaaaaaca aattgaagtt 180
gcagttgccg gtcgcattat gaccaaacgt ggtaaaggta aagcaggctt tgcacatatt 240
caggatgtta ccggtcagat tcagatttat gtgcgtcagg atgatgttgg tgaacagcag 300
tatgaactgt tcaaaattag cgatctgggt gatattgttg gtgttcgtgg caccatgttt 360
aaaaccaaag tgggtgaact gagcattaaa gtgagcagct atgaatttct gaccaaagca 420
ctgcgtccgc tgccggaaaa atatcatggt ctgaaagata ttgaacagcg ttatcgtcag 480
cgctatctgg atctgattat gaatccggaa agcaaaaaaa cctttattac ccgctcactg 540
attatccaga gcatgcgtcg ttatctggat agccgtggat atctggaagt tgaaaccccg 600
atgatgcatg ccgttgccgg tggtgcagca gcacgtccgt ttattacaca tcataatgca 660
ctggatatga ccctgtatat gcgtattgca attgaactgc atctgaaacg tctgattgtt 720
ggcggtctgg aaaaagtgta tgaaattggt cgtgtgtttc gcaatgaagg tattagcacc 780
cgtcataatc cggaatttac catgctggaa ctgtacgaag catatgccga ttttcacgat 840
attatggaac tgaccgaaaa cctgattgcc catattgcaa ccgaagttct gggcaccacc 900
aaaattcagt atgatgaaca tgttgttgac ctgacaccgg aatggcgtcg tctgcatatg 960
gttgatgcaa ttaaagaata tgtcggcgtg gatttttggc gtcagatgag tgatgaagaa 1020
gcacgcgaac tggcaaaaga acatggtgtg gaagttgcac cgcatatgac ctttggccat 1080
attgtgaacg aattctttga gcagaaagtg gaaagccatc tgattcagcc gacctttatc 1140
tatggtcatc cggttgaaat tagtccgctg gccaaaaaaa acccggatga tcctcgtttt 1200
accgatcgtt ttgagctgtt tattgtgggt cgtgaacatg caaatgcctt taccgaactg 1260
aacgatccga ttgatcagcg tcagcgtttt gaagcacagc tgaaagaacg tgaacagggt 1320
aatgatgaag cacacgaaat ggatgaagat tttctggaag cactggaata tggtatgcct 1380
ccgaccggtg gtttaggtat tggtgttgat cgtctggtta tgctgctgac caatagtccg 1440
agcattcgtg atgttctgct gtttccgcag atgcgtcata aataa 1485
<210> 48
<211> 494
<212> PRT
<213> Geobacillus stearothermophilus
<400> 48
Met Ser His Glu Glu Leu Asn Asp Gln Leu Arg Val Arg Arg Glu Lys
1 5 10 15
Leu Lys Lys Ile Glu Glu Leu Gly Val Asp Pro Phe Gly Lys Arg Phe
20 25 30
Glu Arg Thr His Lys Ala Gln Glu Leu Phe Glu Leu Tyr Gly Asp Leu
35 40 45
Ser Lys Glu Glu Leu Glu Glu Lys Gln Ile Glu Val Ala Val Ala Gly
50 55 60
Arg Ile Met Thr Lys Arg Gly Lys Gly Lys Ala Gly Phe Ala His Ile
65 70 75 80
Gln Asp Val Thr Gly Gln Ile Gln Ile Tyr Val Arg Gln Asp Asp Val
85 90 95
Gly Glu Gln Gln Tyr Glu Leu Phe Lys Ile Ser Asp Leu Gly Asp Ile
100 105 110
Val Gly Val Arg Gly Thr Met Phe Lys Thr Lys Val Gly Glu Leu Ser
115 120 125
Ile Lys Val Ser Ser Tyr Glu Phe Leu Thr Lys Ala Leu Arg Pro Leu
130 135 140
Pro Glu Lys Tyr His Gly Leu Lys Asp Ile Glu Gln Arg Tyr Arg Gln
145 150 155 160
Arg Tyr Leu Asp Leu Ile Met Asn Pro Glu Ser Lys Lys Thr Phe Ile
165 170 175
Thr Arg Ser Leu Ile Ile Gln Ser Met Arg Arg Tyr Leu Asp Ser Arg
180 185 190
Gly Tyr Leu Glu Val Glu Thr Pro Met Met His Ala Val Ala Gly Gly
195 200 205
Ala Ala Ala Arg Pro Phe Ile Thr His His Asn Ala Leu Asp Met Thr
210 215 220
Leu Tyr Met Arg Ile Ala Ile Glu Leu His Leu Lys Arg Leu Ile Val
225 230 235 240
Gly Gly Leu Glu Lys Val Tyr Glu Ile Gly Arg Val Phe Arg Asn Glu
245 250 255
Gly Ile Ser Thr Arg His Asn Pro Glu Phe Thr Met Leu Glu Leu Tyr
260 265 270
Glu Ala Tyr Ala Asp Phe His Asp Ile Met Glu Leu Thr Glu Asn Leu
275 280 285
Ile Ala His Ile Ala Thr Glu Val Leu Gly Thr Thr Lys Ile Gln Tyr
290 295 300
Asp Glu His Val Val Asp Leu Thr Pro Glu Trp Arg Arg Leu His Met
305 310 315 320
Val Asp Ala Ile Lys Glu Tyr Val Gly Val Asp Phe Trp Arg Gln Met
325 330 335
Ser Asp Glu Glu Ala Arg Glu Leu Ala Lys Glu His Gly Val Glu Val
340 345 350
Ala Pro His Met Thr Phe Gly His Ile Val Asn Glu Phe Phe Glu Gln
355 360 365
Lys Val Glu Ser His Leu Ile Gln Pro Thr Phe Ile Tyr Gly His Pro
370 375 380
Val Glu Ile Ser Pro Leu Ala Lys Lys Asn Pro Asp Asp Pro Arg Phe
385 390 395 400
Thr Asp Arg Phe Glu Leu Phe Ile Val Gly Arg Glu His Ala Asn Ala
405 410 415
Phe Thr Glu Leu Asn Asp Pro Ile Asp Gln Arg Gln Arg Phe Glu Ala
420 425 430
Gln Leu Lys Glu Arg Glu Gln Gly Asn Asp Glu Ala His Glu Met Asp
435 440 445
Glu Asp Phe Leu Glu Ala Leu Glu Tyr Gly Met Pro Pro Thr Gly Gly
450 455 460
Leu Gly Ile Gly Val Asp Arg Leu Val Met Leu Leu Thr Asn Ser Pro
465 470 475 480
Ser Ile Arg Asp Val Leu Leu Phe Pro Gln Met Arg His Lys
485 490
<210> 49
<211> 1953
<212> DNA
<213> Geobacillus stearothermophilus
<400> 49
atggaaaaaa agaccttcta tctgaccacg ccgatctatt atccgagcga tcgtctgcat 60
attggtcatg catataccac cgttgccggt gatgcaatgg cacgttataa acgtatgcgt 120
ggttatgatg ttatgtatct gaccggcacc gatgaacatg gtcagaaaat tcagcgtaaa 180
gccgaagaaa aaggtgttac accgcagcag tatgttgatg aaattgttgc aggtattcaa 240
gaactgtgga aaaaactgga tatcagctat gatgatttca tccgtaccac acaagaacgc 300
cataaaaaag ttgttgagca gatttttacc cgtctggttg aacagggtga tatttatctg 360
ggtgaatatg aaggttggta ttgtaccccg tgtgaaagct tttataccga acgtcagctg 420
gttgatggta attgtccgga ttgtggtcgt ccggttgaaa aagttaaaga ggaaagctat 480
tttttccgca tgagcaaata tgttgatcgc ctgctgcagt attatgaaga aaacccggat 540
ttcattcagc cggaaagccg taaaaatgag atgattaaca actttatcaa acctggcctg 600
gaagatctgg cagttagccg taccaccttt gattggggta ttaaagttcc gggtaatccg 660
aaacatgtga tctatgtttg gattgatgca ctggccaact atattaccgc attaggttat 720
ggcaccgata acgatgaaaa attccgtaaa tattggcctg ccgatgttca tctggttggt 780
aaagaaattg ttcgcttcca taccatttat tggccgatta tgctgatggc actgggtctg 840
ccgctgccga aaaaagtttt tggtcatggt tggctgctga tgaaagatgg taaaatgagc 900
aaaagcaaag gcaatgttgt tgatccggtt acactgattg atcgttatgg tctggatgca 960
ctgcgttatt atctgctgcg tgaagttccg tttggtgcag atggtgtttt tacaccggaa 1020
ggttttattg agcgcatcaa ttatgatctg gcaaatgatc tgggtaatct gctgcatcgt 1080
accgttgcaa tgatcgaaaa atactttgat ggtgtgattc cgccttatcg tggtccgaaa 1140
acaccgtttg atcaagagct ggttcagacc gcacgtgaag ttgttcgtca gtatgaagag 1200
gcaatggaag gtatggaatt tagcgttgca ctggcagcag tttggcagct gattagtcgt 1260
accaataaat acattgatga aacccagccg tgggtgttag caaaagatga acagaaacgt 1320
gatgaactgg cagccgttat gacccatctg gcagaaagcc tgcgtcatac cgcagttctg 1380
ctgcagccgt ttctgacccg cacaccggaa cgtatgctgg cacagctggg tattaccgat 1440
catagcctga aagaatggga tagcctgtat gattttggtc tgattccgga aggcaccaaa 1500
gttcagaaag gtgaaccgct gtttccgcgt ctggatattg aagcagaagt ggaatatatc 1560
aaagcccata tgcaaggtgg taaaccggca gccgaaccgg ttaaagaaga aaaaaaagca 1620
gccgaagcag cggaaattag catcgatgaa tttgcaaaag ttgatctgcg tgttgccgaa 1680
gttattcatg cagaacgtat gaaaaacgcc gataaactgc tgaaactgca gctggattta 1740
ggtggtgaaa aacgtcaggt tattagcggt attgccgaat tctataaacc ggaagaactg 1800
gtgggtaaaa aagtgatttg tgtggcaaat ctgaaaccgg caaaactgcg tggtgaatgg 1860
tctgaaggca tgattctggc aggcggtagc ggtggtgaat ttagcctggc aaccgttgat 1920
cagcatgttc cgaatggtac gaaaatcaaa taa 1953
<210> 50
<211> 650
<212> PRT
<213> Geobacillus stearothermophilus
<400> 50
Met Glu Lys Lys Thr Phe Tyr Leu Thr Thr Pro Ile Tyr Tyr Pro Ser
1 5 10 15
Asp Arg Leu His Ile Gly His Ala Tyr Thr Thr Val Ala Gly Asp Ala
20 25 30
Met Ala Arg Tyr Lys Arg Met Arg Gly Tyr Asp Val Met Tyr Leu Thr
35 40 45
Gly Thr Asp Glu His Gly Gln Lys Ile Gln Arg Lys Ala Glu Glu Lys
50 55 60
Gly Val Thr Pro Gln Gln Tyr Val Asp Glu Ile Val Ala Gly Ile Gln
65 70 75 80
Glu Leu Trp Lys Lys Leu Asp Ile Ser Tyr Asp Asp Phe Ile Arg Thr
85 90 95
Thr Gln Glu Arg His Lys Lys Val Val Glu Gln Ile Phe Thr Arg Leu
100 105 110
Val Glu Gln Gly Asp Ile Tyr Leu Gly Glu Tyr Glu Gly Trp Tyr Cys
115 120 125
Thr Pro Cys Glu Ser Phe Tyr Thr Glu Arg Gln Leu Val Asp Gly Asn
130 135 140
Cys Pro Asp Cys Gly Arg Pro Val Glu Lys Val Lys Glu Glu Ser Tyr
145 150 155 160
Phe Phe Arg Met Ser Lys Tyr Val Asp Arg Leu Leu Gln Tyr Tyr Glu
165 170 175
Glu Asn Pro Asp Phe Ile Gln Pro Glu Ser Arg Lys Asn Glu Met Ile
180 185 190
Asn Asn Phe Ile Lys Pro Gly Leu Glu Asp Leu Ala Val Ser Arg Thr
195 200 205
Thr Phe Asp Trp Gly Ile Lys Val Pro Gly Asn Pro Lys His Val Ile
210 215 220
Tyr Val Trp Ile Asp Ala Leu Ala Asn Tyr Ile Thr Ala Leu Gly Tyr
225 230 235 240
Gly Thr Asp Asn Asp Glu Lys Phe Arg Lys Tyr Trp Pro Ala Asp Val
245 250 255
His Leu Val Gly Lys Glu Ile Val Arg Phe His Thr Ile Tyr Trp Pro
260 265 270
Ile Met Leu Met Ala Leu Gly Leu Pro Leu Pro Lys Lys Val Phe Gly
275 280 285
His Gly Trp Leu Leu Met Lys Asp Gly Lys Met Ser Lys Ser Lys Gly
290 295 300
Asn Val Val Asp Pro Val Thr Leu Ile Asp Arg Tyr Gly Leu Asp Ala
305 310 315 320
Leu Arg Tyr Tyr Leu Leu Arg Glu Val Pro Phe Gly Ala Asp Gly Val
325 330 335
Phe Thr Pro Glu Gly Phe Ile Glu Arg Ile Asn Tyr Asp Leu Ala Asn
340 345 350
Asp Leu Gly Asn Leu Leu His Arg Thr Val Ala Met Ile Glu Lys Tyr
355 360 365
Phe Asp Gly Val Ile Pro Pro Tyr Arg Gly Pro Lys Thr Pro Phe Asp
370 375 380
Gln Glu Leu Val Gln Thr Ala Arg Glu Val Val Arg Gln Tyr Glu Glu
385 390 395 400
Ala Met Glu Gly Met Glu Phe Ser Val Ala Leu Ala Ala Val Trp Gln
405 410 415
Leu Ile Ser Arg Thr Asn Lys Tyr Ile Asp Glu Thr Gln Pro Trp Val
420 425 430
Leu Ala Lys Asp Glu Gln Lys Arg Asp Glu Leu Ala Ala Val Met Thr
435 440 445
His Leu Ala Glu Ser Leu Arg His Thr Ala Val Leu Leu Gln Pro Phe
450 455 460
Leu Thr Arg Thr Pro Glu Arg Met Leu Ala Gln Leu Gly Ile Thr Asp
465 470 475 480
His Ser Leu Lys Glu Trp Asp Ser Leu Tyr Asp Phe Gly Leu Ile Pro
485 490 495
Glu Gly Thr Lys Val Gln Lys Gly Glu Pro Leu Phe Pro Arg Leu Asp
500 505 510
Ile Glu Ala Glu Val Glu Tyr Ile Lys Ala His Met Gln Gly Gly Lys
515 520 525
Pro Ala Ala Glu Pro Val Lys Glu Glu Lys Lys Ala Ala Glu Ala Ala
530 535 540
Glu Ile Ser Ile Asp Glu Phe Ala Lys Val Asp Leu Arg Val Ala Glu
545 550 555 560
Val Ile His Ala Glu Arg Met Lys Asn Ala Asp Lys Leu Leu Lys Leu
565 570 575
Gln Leu Asp Leu Gly Gly Glu Lys Arg Gln Val Ile Ser Gly Ile Ala
580 585 590
Glu Phe Tyr Lys Pro Glu Glu Leu Val Gly Lys Lys Val Ile Cys Val
595 600 605
Ala Asn Leu Lys Pro Ala Lys Leu Arg Gly Glu Trp Ser Glu Gly Met
610 615 620
Ile Leu Ala Gly Gly Ser Gly Gly Glu Phe Ser Leu Ala Thr Val Asp
625 630 635 640
Gln His Val Pro Asn Gly Thr Lys Ile Lys
645 650
<210> 51
<211> 1035
<212> DNA
<213> Geobacillus
<400> 51
atgaaagaac gcctgtatga actgaaacgt caggcactgg aacaaattgg tcaggcacgt 60
gatctgcgta tgctgaatga tgttcgtgtt gcatatctgg gtaaaaaagg tccgattacc 120
gaagttctgc gtggtatggg tgcactgcct ccggaagaac gtccgaaaat tggtgcactg 180
gcaaatgaag ttcgtgaagc aattcagcag gccctggaag caaaacaggc aaaacttgaa 240
caagaagaag tggaacgtaa actggcagcc gaagcaattg atgttaccct gcctggtcgt 300
ccggttagcc tgggtaatcc gcatccgctg acacgtgtta ttgaagaaat tgaggacctg 360
tttattggca tgggttatac cgttgcagaa ggtccggaag ttgaaaccga ttattacaat 420
tttgaagccc tgaatctgcc gaaaggtcat ccggcacgcg atatgcagga tagcttttat 480
atcaccgaag aaattctgct gcgtacccat acctcaccga tgcaggcacg taccatggaa 540
aaacatcgtg gtcgtggtcc ggttaaaatc atttgtccgg gtaaagttta tcgtcgcgat 600
accgatgatg caacccatag ccatcagttt acacagattg aaggtctggt tgtggatcgt 660
aatattcgta tgagcgatct gaaaggcacc ctgcgtgaat ttgcccgtaa actgtttggt 720
gaaggtcgtg atattcgttt tcgtccgagc ttttttccgt ttaccgaacc gagcgttgaa 780
gttgatgtta gctgttttcg ttgtgaaggc cgtggttgcg gtgtttgtaa aggcaccggt 840
tggattgaaa ttttaggtgc aggtatggtt catccgaatg ttctggaaat ggcaggtttt 900
gatagtaaaa cctataccgg ttttgcattc ggtatgggtc ctgaacgtat tgcaatgctg 960
aaatatggca ttgatgatat ccgccacttc tatcagaatg atctgcgctt tctgcgtcag 1020
tttctgcgtg tttaa 1035
<210> 52
<211> 344
<212> PRT
<213> Geobacillus
<400> 52
Met Lys Glu Arg Leu Tyr Glu Leu Lys Arg Gln Ala Leu Glu Gln Ile
1 5 10 15
Gly Gln Ala Arg Asp Leu Arg Met Leu Asn Asp Val Arg Val Ala Tyr
20 25 30
Leu Gly Lys Lys Gly Pro Ile Thr Glu Val Leu Arg Gly Met Gly Ala
35 40 45
Leu Pro Pro Glu Glu Arg Pro Lys Ile Gly Ala Leu Ala Asn Glu Val
50 55 60
Arg Glu Ala Ile Gln Gln Ala Leu Glu Ala Lys Gln Ala Lys Leu Glu
65 70 75 80
Gln Glu Glu Val Glu Arg Lys Leu Ala Ala Glu Ala Ile Asp Val Thr
85 90 95
Leu Pro Gly Arg Pro Val Ser Leu Gly Asn Pro His Pro Leu Thr Arg
100 105 110
Val Ile Glu Glu Ile Glu Asp Leu Phe Ile Gly Met Gly Tyr Thr Val
115 120 125
Ala Glu Gly Pro Glu Val Glu Thr Asp Tyr Tyr Asn Phe Glu Ala Leu
130 135 140
Asn Leu Pro Lys Gly His Pro Ala Arg Asp Met Gln Asp Ser Phe Tyr
145 150 155 160
Ile Thr Glu Glu Ile Leu Leu Arg Thr His Thr Ser Pro Met Gln Ala
165 170 175
Arg Thr Met Glu Lys His Arg Gly Arg Gly Pro Val Lys Ile Ile Cys
180 185 190
Pro Gly Lys Val Tyr Arg Arg Asp Thr Asp Asp Ala Thr His Ser His
195 200 205
Gln Phe Thr Gln Ile Glu Gly Leu Val Val Asp Arg Asn Ile Arg Met
210 215 220
Ser Asp Leu Lys Gly Thr Leu Arg Glu Phe Ala Arg Lys Leu Phe Gly
225 230 235 240
Glu Gly Arg Asp Ile Arg Phe Arg Pro Ser Phe Phe Pro Phe Thr Glu
245 250 255
Pro Ser Val Glu Val Asp Val Ser Cys Phe Arg Cys Glu Gly Arg Gly
260 265 270
Cys Gly Val Cys Lys Gly Thr Gly Trp Ile Glu Ile Leu Gly Ala Gly
275 280 285
Met Val His Pro Asn Val Leu Glu Met Ala Gly Phe Asp Ser Lys Thr
290 295 300
Tyr Thr Gly Phe Ala Phe Gly Met Gly Pro Glu Arg Ile Ala Met Leu
305 310 315 320
Lys Tyr Gly Ile Asp Asp Ile Arg His Phe Tyr Gln Asn Asp Leu Arg
325 330 335
Phe Leu Arg Gln Phe Leu Arg Val
340
<210> 53
<211> 2415
<212> DNA
<213> Geobacillus stearothermophilus
<400> 53
atgctggtta gctatcgttg gctgggtgaa tatgttgatc tgaccggtat taccgcaaaa 60
gaactggcag aacgtattac caaaagcggt attgaagttg aacgtgttga agcactggat 120
cgtggtatga atggtgttgt tattggtcat gttctggaat gtgaaccgca tccgaatgca 180
gataaactgc gtaaatgtct ggttgattta ggtgaaggtg aaccggtgcg tattatttgt 240
ggtgcaccga atgttgcaaa aggtcagaaa gttgcagttg ccaaagttgg tgcagttctg 300
cctggtaact ttaaaatcaa acgtgcaaaa ctgcgtggcg aagaaagcaa tggtatgatt 360
tgtagcctgc aagaactggg tgttgaaacc aaagttgttc cgaaagaata tgccgatggc 420
atttttgttt ttccgagtga tgcaccggtt ggtgccgatg cactggaatg gctgggtctg 480
catgatgaag ttctggaact ggcactgacc ccgaatcgtg cagattgtct gagcatgatt 540
ggtgttgcct atgaagttgc agcaattctg ggtcgtgatg ttaaactgcc ggaagcagca 600
gttaaagaaa atagcgaaca tgtgcacgaa tatatcagcg ttcgtgtgga agcaccggaa 660
gataatccgc tgtatgcagg tcgtattgtt aaaaatgttc gtattggtcc gagtccgctg 720
tggatgcagg cacgtctgat ggcagcaggt attcgtccgc ataataatgt tgttgacatc 780
accaactata tcctgctgga atatggtcag ccgctgcatg catttgatta tgatcgtctg 840
ggtagcaaag aaattgttgt tcgtcgtgca aaagccggtg aaaccattat taccctggat 900
gatgttgaac gtaaactgac cgaaaatcat ctggtgatta ccaatggtcg cgaaccggtt 960
gcactggcag gcgttatggg tggtgccaat agcgaagttc gtgatgatac caccaccgtt 1020
tttattgaag cagcctattt caccagtccg gttattcgtc aggccgttaa agatcatggt 1080
ctgcgtagcg aagcgagcac ccgttttgaa aaaggtattg atccggcacg taccaaagag 1140
gccctggatc gcgcagcagc actgatgagc gaatatgcag gcggtgaagt tgttggtggt 1200
attgttgaag ccagcgtttg gcgtcaggat ccggttgttg ttaccgttac actggaacgc 1260
attaatggtg ttctgggcac cgcaatgacc aaagaagaag tggctgccat tctgagcaat 1320
ctgcagtttc cgtttaccga agataatggc acctttacca ttcatgttcc gagccgtcgt 1380
cgtgatattg caattgaaga agatattatt gaagaggcag cccgtctgta tggttatgat 1440
cgcctgcctg caacactgcc ggttgccgaa gcaaaacctg gtggtctgac accgcatcag 1500
gcaaaacgtc gtcgcgttcg tcgttatctg gaaggcaccg gtctgtttca ggcaattacc 1560
tatagcctga cctcaccgga taaagcaacc cgctttgccc tggaaaccgc agaaccgatt 1620
cgtctggcac tgccgatgag tgaagaacgt agcgttctgc gtcagagcct gattccgcat 1680
ctgctggaag ccgcaagcta taatcgtgca cgtcaggttg aagatgttgc cctgtatgaa 1740
attggtagcg tttatctgag caaaggtgaa catgtacagc ctgcagaaaa agaacgttta 1800
gccggtgtgc tgacaggtct gtggcatgca catctgtggc agggtgaaaa aaaagccgtt 1860
gatttttatg tggccaaagg tattctggat ggtctgtttg atctgctggg tttagcagca 1920
cgtattgaat ataaaccggc aaaacgcgct gatctgcatc cgggtcgtac cgcagatatt 1980
gtgctggatg gccgtgtgat tggttttgtt ggtcagctgc atcctgcagt tcagaaagag 2040
tatgatctga aagaaaccta tgtgtttgag ctggccctga ccgatctgct gaatgcagaa 2100
agcgaagcaa ttcgttatga acctattccg cgttttccga gcgttgtgcg cgacattgca 2160
ctggttgttg atgaaaatgt tgaagcgggt gcactgaaac aggcaatcga agaagcaggt 2220
aaaccgctgg ttaaagatgt tagcctgttc gatgtttata aaggcgatcg tctgccggat 2280
ggtaaaaaaa gtctggcatt tagcctgcgt tattatgatc cggaacgcac cctgacagat 2340
gaagaggttg cagcagtgca tgaacgtgtg ctggcagcag ttgaaaaaca gtttggtgcc 2400
gtgctgcgtg gttaa 2415
<210> 54
<211> 804
<212> PRT
<213> Geobacillus stearothermophilus
<400> 54
Met Leu Val Ser Tyr Arg Trp Leu Gly Glu Tyr Val Asp Leu Thr Gly
1 5 10 15
Ile Thr Ala Lys Glu Leu Ala Glu Arg Ile Thr Lys Ser Gly Ile Glu
20 25 30
Val Glu Arg Val Glu Ala Leu Asp Arg Gly Met Asn Gly Val Val Ile
35 40 45
Gly His Val Leu Glu Cys Glu Pro His Pro Asn Ala Asp Lys Leu Arg
50 55 60
Lys Cys Leu Val Asp Leu Gly Glu Gly Glu Pro Val Arg Ile Ile Cys
65 70 75 80
Gly Ala Pro Asn Val Ala Lys Gly Gln Lys Val Ala Val Ala Lys Val
85 90 95
Gly Ala Val Leu Pro Gly Asn Phe Lys Ile Lys Arg Ala Lys Leu Arg
100 105 110
Gly Glu Glu Ser Asn Gly Met Ile Cys Ser Leu Gln Glu Leu Gly Val
115 120 125
Glu Thr Lys Val Val Pro Lys Glu Tyr Ala Asp Gly Ile Phe Val Phe
130 135 140
Pro Ser Asp Ala Pro Val Gly Ala Asp Ala Leu Glu Trp Leu Gly Leu
145 150 155 160
His Asp Glu Val Leu Glu Leu Ala Leu Thr Pro Asn Arg Ala Asp Cys
165 170 175
Leu Ser Met Ile Gly Val Ala Tyr Glu Val Ala Ala Ile Leu Gly Arg
180 185 190
Asp Val Lys Leu Pro Glu Ala Ala Val Lys Glu Asn Ser Glu His Val
195 200 205
His Glu Tyr Ile Ser Val Arg Val Glu Ala Pro Glu Asp Asn Pro Leu
210 215 220
Tyr Ala Gly Arg Ile Val Lys Asn Val Arg Ile Gly Pro Ser Pro Leu
225 230 235 240
Trp Met Gln Ala Arg Leu Met Ala Ala Gly Ile Arg Pro His Asn Asn
245 250 255
Val Val Asp Ile Thr Asn Tyr Ile Leu Leu Glu Tyr Gly Gln Pro Leu
260 265 270
His Ala Phe Asp Tyr Asp Arg Leu Gly Ser Lys Glu Ile Val Val Arg
275 280 285
Arg Ala Lys Ala Gly Glu Thr Ile Ile Thr Leu Asp Asp Val Glu Arg
290 295 300
Lys Leu Thr Glu Asn His Leu Val Ile Thr Asn Gly Arg Glu Pro Val
305 310 315 320
Ala Leu Ala Gly Val Met Gly Gly Ala Asn Ser Glu Val Arg Asp Asp
325 330 335
Thr Thr Thr Val Phe Ile Glu Ala Ala Tyr Phe Thr Ser Pro Val Ile
340 345 350
Arg Gln Ala Val Lys Asp His Gly Leu Arg Ser Glu Ala Ser Thr Arg
355 360 365
Phe Glu Lys Gly Ile Asp Pro Ala Arg Thr Lys Glu Ala Leu Asp Arg
370 375 380
Ala Ala Ala Leu Met Ser Glu Tyr Ala Gly Gly Glu Val Val Gly Gly
385 390 395 400
Ile Val Glu Ala Ser Val Trp Arg Gln Asp Pro Val Val Val Thr Val
405 410 415
Thr Leu Glu Arg Ile Asn Gly Val Leu Gly Thr Ala Met Thr Lys Glu
420 425 430
Glu Val Ala Ala Ile Leu Ser Asn Leu Gln Phe Pro Phe Thr Glu Asp
435 440 445
Asn Gly Thr Phe Thr Ile His Val Pro Ser Arg Arg Arg Asp Ile Ala
450 455 460
Ile Glu Glu Asp Ile Ile Glu Glu Ala Ala Arg Leu Tyr Gly Tyr Asp
465 470 475 480
Arg Leu Pro Ala Thr Leu Pro Val Ala Glu Ala Lys Pro Gly Gly Leu
485 490 495
Thr Pro His Gln Ala Lys Arg Arg Arg Val Arg Arg Tyr Leu Glu Gly
500 505 510
Thr Gly Leu Phe Gln Ala Ile Thr Tyr Ser Leu Thr Ser Pro Asp Lys
515 520 525
Ala Thr Arg Phe Ala Leu Glu Thr Ala Glu Pro Ile Arg Leu Ala Leu
530 535 540
Pro Met Ser Glu Glu Arg Ser Val Leu Arg Gln Ser Leu Ile Pro His
545 550 555 560
Leu Leu Glu Ala Ala Ser Tyr Asn Arg Ala Arg Gln Val Glu Asp Val
565 570 575
Ala Leu Tyr Glu Ile Gly Ser Val Tyr Leu Ser Lys Gly Glu His Val
580 585 590
Gln Pro Ala Glu Lys Glu Arg Leu Ala Gly Val Leu Thr Gly Leu Trp
595 600 605
His Ala His Leu Trp Gln Gly Glu Lys Lys Ala Val Asp Phe Tyr Val
610 615 620
Ala Lys Gly Ile Leu Asp Gly Leu Phe Asp Leu Leu Gly Leu Ala Ala
625 630 635 640
Arg Ile Glu Tyr Lys Pro Ala Lys Arg Ala Asp Leu His Pro Gly Arg
645 650 655
Thr Ala Asp Ile Val Leu Asp Gly Arg Val Ile Gly Phe Val Gly Gln
660 665 670
Leu His Pro Ala Val Gln Lys Glu Tyr Asp Leu Lys Glu Thr Tyr Val
675 680 685
Phe Glu Leu Ala Leu Thr Asp Leu Leu Asn Ala Glu Ser Glu Ala Ile
690 695 700
Arg Tyr Glu Pro Ile Pro Arg Phe Pro Ser Val Val Arg Asp Ile Ala
705 710 715 720
Leu Val Val Asp Glu Asn Val Glu Ala Gly Ala Leu Lys Gln Ala Ile
725 730 735
Glu Glu Ala Gly Lys Pro Leu Val Lys Asp Val Ser Leu Phe Asp Val
740 745 750
Tyr Lys Gly Asp Arg Leu Pro Asp Gly Lys Lys Ser Leu Ala Phe Ser
755 760 765
Leu Arg Tyr Tyr Asp Pro Glu Arg Thr Leu Thr Asp Glu Glu Val Ala
770 775 780
Ala Val His Glu Arg Val Leu Ala Ala Val Glu Lys Gln Phe Gly Ala
785 790 795 800
Val Leu Arg Gly
<210> 55
<211> 1704
<212> DNA
<213> Geobacillus stearothermophilus
<400> 55
atgcgtcaga gccaggcatt tattccgaca ctgcgtgaag ttccggcaga tgcagaagtt 60
aaaagccatc agctgctgct gcgtgcaggt tttattcgtc agagcgcaag cggtgtttat 120
acctttctgc cgctgggtca gcgtgtgctg cagaaagttg aagcaattat tcgcgaagaa 180
atgaatcgta ttggtgccat ggaactgttt atgcctgcac tgcagcctgc agaactgtgg 240
cagcagagcg gtcgttggta tagctatggt ccggaactga tgcgtctgaa agatcgtcat 300
gaacgtgatt ttgcactggg tccgacacat gaagagatga ttaccgcaat tgttcgtgat 360
gaggtgaaaa cctataaacg tctgcctctg gttctgtatc agatccagac caaattccgt 420
gatgaaaaac gtccgcgttt tggtctgtta cgtggtcgtg aatttatgat gaaagatgcc 480
tatagcttcc ataccagcaa agaaagcctg gatgaaacct acaacaatat gtatgaagcc 540
tacgccaaca tttttcgtcg ttgcggtctg aattttcgtg cagttattgc agatagcggt 600
gcaattggtg gtaaagatac ccacgaattc atggttctga gcgatattgg tgaagatacc 660
attgcatata gtgatgcaag cgattatgca gccaatattg aaatggcacc ggttgttgca 720
acctatgaaa aaagtgatga acctccggca gaactgaaga aagttgccac accgggtcag 780
aaaaccattg ccgaagttgc aagccatctg caaattagtc cggaacgttg tattaaaagc 840
ctgctgttta atgtggatgg tcgttatgtt ctggtgctgg ttcgtggtga tcatgaagca 900
aatgaagtga aagtgaaaaa tgtgctggat gccaccgttg ttgaactggc aaaaccggaa 960
gaaaccgaac gtgttatgaa tgcaccgatt ggtagcctgg gtcctattgg tgttagcgaa 1020
gatgttaccg ttattgccga tcatgcagtt gcagcaattg ttaatggtgt ttgtggtgcc 1080
aatgaagagg gctatcatta cattggtgtg aatccgggtc gcgattttgc agttagccag 1140
tatgccgatc tgcgttttgt taaagaaggt gatccgagtc cggatggtaa aggcaccatt 1200
cgttttgcac gtggtattga agttggccat gtttttaaac tgggcaccaa atatagcgaa 1260
gccatgaatg cagtttatct ggatgagaat ggtcagaccc agacaatgat tatgggttgt 1320
tatggtattg gcgttagccg tctggttgca gccattgcag aacagtttgc cgatgaacat 1380
ggtctggttt ggcctgcaag cgttgcaccg tttcatattc atctgctgac cgcaaatgcc 1440
aaatcagatg aacagcgtgc actggccgaa gaatggtatg aaaaactggg tcaagcaggt 1500
tttgaagtgc tgtatgatga tcgtccagaa cgtgccggtg ttaaatttgc cgatagcgat 1560
ctgattggta ttccgctgcg tgttaccgtg ggtaaacgtg caggcgaagg tgttgttgaa 1620
gttaaagttc gtaaaaccgg tgaaaccttt gatgttccgg ttagcgaact ggttgatacc 1680
gcacgtcgtc tgctgcagag ctaa 1704
<210> 56
<211> 567
<212> PRT
<213> Geobacillus stearothermophilus
<400> 56
Met Arg Gln Ser Gln Ala Phe Ile Pro Thr Leu Arg Glu Val Pro Ala
1 5 10 15
Asp Ala Glu Val Lys Ser His Gln Leu Leu Leu Arg Ala Gly Phe Ile
20 25 30
Arg Gln Ser Ala Ser Gly Val Tyr Thr Phe Leu Pro Leu Gly Gln Arg
35 40 45
Val Leu Gln Lys Val Glu Ala Ile Ile Arg Glu Glu Met Asn Arg Ile
50 55 60
Gly Ala Met Glu Leu Phe Met Pro Ala Leu Gln Pro Ala Glu Leu Trp
65 70 75 80
Gln Gln Ser Gly Arg Trp Tyr Ser Tyr Gly Pro Glu Leu Met Arg Leu
85 90 95
Lys Asp Arg His Glu Arg Asp Phe Ala Leu Gly Pro Thr His Glu Glu
100 105 110
Met Ile Thr Ala Ile Val Arg Asp Glu Val Lys Thr Tyr Lys Arg Leu
115 120 125
Pro Leu Val Leu Tyr Gln Ile Gln Thr Lys Phe Arg Asp Glu Lys Arg
130 135 140
Pro Arg Phe Gly Leu Leu Arg Gly Arg Glu Phe Met Met Lys Asp Ala
145 150 155 160
Tyr Ser Phe His Thr Ser Lys Glu Ser Leu Asp Glu Thr Tyr Asn Asn
165 170 175
Met Tyr Glu Ala Tyr Ala Asn Ile Phe Arg Arg Cys Gly Leu Asn Phe
180 185 190
Arg Ala Val Ile Ala Asp Ser Gly Ala Ile Gly Gly Lys Asp Thr His
195 200 205
Glu Phe Met Val Leu Ser Asp Ile Gly Glu Asp Thr Ile Ala Tyr Ser
210 215 220
Asp Ala Ser Asp Tyr Ala Ala Asn Ile Glu Met Ala Pro Val Val Ala
225 230 235 240
Thr Tyr Glu Lys Ser Asp Glu Pro Pro Ala Glu Leu Lys Lys Val Ala
245 250 255
Thr Pro Gly Gln Lys Thr Ile Ala Glu Val Ala Ser His Leu Gln Ile
260 265 270
Ser Pro Glu Arg Cys Ile Lys Ser Leu Leu Phe Asn Val Asp Gly Arg
275 280 285
Tyr Val Leu Val Leu Val Arg Gly Asp His Glu Ala Asn Glu Val Lys
290 295 300
Val Lys Asn Val Leu Asp Ala Thr Val Val Glu Leu Ala Lys Pro Glu
305 310 315 320
Glu Thr Glu Arg Val Met Asn Ala Pro Ile Gly Ser Leu Gly Pro Ile
325 330 335
Gly Val Ser Glu Asp Val Thr Val Ile Ala Asp His Ala Val Ala Ala
340 345 350
Ile Val Asn Gly Val Cys Gly Ala Asn Glu Glu Gly Tyr His Tyr Ile
355 360 365
Gly Val Asn Pro Gly Arg Asp Phe Ala Val Ser Gln Tyr Ala Asp Leu
370 375 380
Arg Phe Val Lys Glu Gly Asp Pro Ser Pro Asp Gly Lys Gly Thr Ile
385 390 395 400
Arg Phe Ala Arg Gly Ile Glu Val Gly His Val Phe Lys Leu Gly Thr
405 410 415
Lys Tyr Ser Glu Ala Met Asn Ala Val Tyr Leu Asp Glu Asn Gly Gln
420 425 430
Thr Gln Thr Met Ile Met Gly Cys Tyr Gly Ile Gly Val Ser Arg Leu
435 440 445
Val Ala Ala Ile Ala Glu Gln Phe Ala Asp Glu His Gly Leu Val Trp
450 455 460
Pro Ala Ser Val Ala Pro Phe His Ile His Leu Leu Thr Ala Asn Ala
465 470 475 480
Lys Ser Asp Glu Gln Arg Ala Leu Ala Glu Glu Trp Tyr Glu Lys Leu
485 490 495
Gly Gln Ala Gly Phe Glu Val Leu Tyr Asp Asp Arg Pro Glu Arg Ala
500 505 510
Gly Val Lys Phe Ala Asp Ser Asp Leu Ile Gly Ile Pro Leu Arg Val
515 520 525
Thr Val Gly Lys Arg Ala Gly Glu Gly Val Val Glu Val Lys Val Arg
530 535 540
Lys Thr Gly Glu Thr Phe Asp Val Pro Val Ser Glu Leu Val Asp Thr
545 550 555 560
Ala Arg Arg Leu Leu Gln Ser
565
<210> 57
<211> 1275
<212> DNA
<213> Geobacillus stearothermophilus
<400> 57
atgctggatg tgaaaattct gcgtacccag tttgaagagg tgaaagaaaa actgatgcag 60
cgtggtggtg atctgaccaa tattgatcgt tttgaacagc tggataaaga tcgtcgtcgt 120
ctgattgcag aagttgaaga actgaaaagc aaacgcaatg atgttagcca gcagattgca 180
gttctgaaac gcgaaaaaaa agatgcagaa ccgctgattg cacagatgcg tgaagttggt 240
gatcgtatta aacgtatgga tgagcagatt cgtcagctgg aagcagaact ggatgatctg 300
ctgctgagca ttccgaatgt tccgcatgaa agcgttccga ttggccagag cgaagaagat 360
aacgttgaag ttcgtcgttg gggtgaaccg cgtagcttta gctttgaacc gaaaccgcat 420
tgggaaattg cagatcgtct gggtctgctg gattttgaac gtgcagcaaa agttgcaggt 480
agccgttttg ttttctataa aggtctgggt gcacgtctgg aacgtgcact gattaacttt 540
atgctggata ttcacctgga tgagtttggc tatgaagaag ttctgcctcc gtatctggtt 600
aatcgtgcaa gcatgattgg caccggtcag ctgccgaaat ttgcagaaga tgcatttcat 660
ctggatagcg aggattattt tctgattccg accgcagaag ttccggttac caatctgcat 720
cgtgatgaaa ttctggcagc agatgacctg ccgatctatt atgcagcata tagcgcatgt 780
tttcgtgcag aagcaggtag cgcaggtcgt gatacccgtg gtctgattcg ccagcatcag 840
ttcaataaag ttgaactggt gaaattcgtg aagccggaag atagctatga tgaactggaa 900
aagctgaccc gtcaggcaga aaccattctg cagcgtctgg gcctgccgta tcgtgttgtt 960
gcactgtgta ccggtgatct gggttttagc gttgcaaaaa cctatgatat tgaagtttgg 1020
ctgccgagct atggcaccta tcgtgaaatt agcagctgta gcaattttga agcatttcag 1080
gcacgtcgtg ccaatattcg ttttcgtcgt gatccgaaag caaaaccgga atatgttcat 1140
accctgaatg gtagcggtct ggcaattggt cgtaccgttg cagcaattct ggaaaattat 1200
cagcaagaag atggcagcgt tattgttccg gaagcactgc gtccgtatat gggcaatcgt 1260
gatgttattc gttaa 1275
<210> 58
<211> 424
<212> PRT
<213> Geobacillus stearothermophilus
<400> 58
Met Leu Asp Val Lys Ile Leu Arg Thr Gln Phe Glu Glu Val Lys Glu
1 5 10 15
Lys Leu Met Gln Arg Gly Gly Asp Leu Thr Asn Ile Asp Arg Phe Glu
20 25 30
Gln Leu Asp Lys Asp Arg Arg Arg Leu Ile Ala Glu Val Glu Glu Leu
35 40 45
Lys Ser Lys Arg Asn Asp Val Ser Gln Gln Ile Ala Val Leu Lys Arg
50 55 60
Glu Lys Lys Asp Ala Glu Pro Leu Ile Ala Gln Met Arg Glu Val Gly
65 70 75 80
Asp Arg Ile Lys Arg Met Asp Glu Gln Ile Arg Gln Leu Glu Ala Glu
85 90 95
Leu Asp Asp Leu Leu Leu Ser Ile Pro Asn Val Pro His Glu Ser Val
100 105 110
Pro Ile Gly Gln Ser Glu Glu Asp Asn Val Glu Val Arg Arg Trp Gly
115 120 125
Glu Pro Arg Ser Phe Ser Phe Glu Pro Lys Pro His Trp Glu Ile Ala
130 135 140
Asp Arg Leu Gly Leu Leu Asp Phe Glu Arg Ala Ala Lys Val Ala Gly
145 150 155 160
Ser Arg Phe Val Phe Tyr Lys Gly Leu Gly Ala Arg Leu Glu Arg Ala
165 170 175
Leu Ile Asn Phe Met Leu Asp Ile His Leu Asp Glu Phe Gly Tyr Glu
180 185 190
Glu Val Leu Pro Pro Tyr Leu Val Asn Arg Ala Ser Met Ile Gly Thr
195 200 205
Gly Gln Leu Pro Lys Phe Ala Glu Asp Ala Phe His Leu Asp Ser Glu
210 215 220
Asp Tyr Phe Leu Ile Pro Thr Ala Glu Val Pro Val Thr Asn Leu His
225 230 235 240
Arg Asp Glu Ile Leu Ala Ala Asp Asp Leu Pro Ile Tyr Tyr Ala Ala
245 250 255
Tyr Ser Ala Cys Phe Arg Ala Glu Ala Gly Ser Ala Gly Arg Asp Thr
260 265 270
Arg Gly Leu Ile Arg Gln His Gln Phe Asn Lys Val Glu Leu Val Lys
275 280 285
Phe Val Lys Pro Glu Asp Ser Tyr Asp Glu Leu Glu Lys Leu Thr Arg
290 295 300
Gln Ala Glu Thr Ile Leu Gln Arg Leu Gly Leu Pro Tyr Arg Val Val
305 310 315 320
Ala Leu Cys Thr Gly Asp Leu Gly Phe Ser Val Ala Lys Thr Tyr Asp
325 330 335
Ile Glu Val Trp Leu Pro Ser Tyr Gly Thr Tyr Arg Glu Ile Ser Ser
340 345 350
Cys Ser Asn Phe Glu Ala Phe Gln Ala Arg Arg Ala Asn Ile Arg Phe
355 360 365
Arg Arg Asp Pro Lys Ala Lys Pro Glu Tyr Val His Thr Leu Asn Gly
370 375 380
Ser Gly Leu Ala Ile Gly Arg Thr Val Ala Ala Ile Leu Glu Asn Tyr
385 390 395 400
Gln Gln Glu Asp Gly Ser Val Ile Val Pro Glu Ala Leu Arg Pro Tyr
405 410 415
Met Gly Asn Arg Asp Val Ile Arg
420
<210> 59
<211> 1959
<212> DNA
<213> Geobacillus
<400> 59
atgccggatg ttattcgtat tacctttccg gatggtgccg aaaaagaatt tccgaaaggc 60
accaccaccg aagatgttgc agcaagcatt agtccgggtc tgaaaaaaaa ggcaattgcg 120
ggtaaactga atggtcgttt tgttgatctg cgtacaccgc tgcatgaaga tggtgaactg 180
gtgattatta cccaggatat gccggaagca ctggatattc tgcgtcatag caccgcacat 240
ctgatggcac aggcaattaa acgtctgtat ggcaatgtga aattaggtgt tggtccggtg 300
attgaaaacg gcttctatta tgatatcgac atggaacata aactgacacc ggatgatctg 360
ccgaaaattg aagcagaaat gcgcaaaatc gtgaaagaga acctggatat tgttcgcaaa 420
gaagttagtc gcgaagaggc aattcgcctg tatgaagaaa ttggtgatga actgaaactg 480
gaactgattg cagatattcc ggaaggtgaa ccgattagca tttatgaaca gggcgaattt 540
tttgatctgt gccgtggtgt tcatgttccg agcaccggta aaatcaaaga atttaaactg 600
ctgagcatca gcggtgcata ttggcgtggt gatagcaata acaaaatgct gcagcgtatt 660
tatggcaccg cgtttttcaa aaaagaagat ctggatcgtt atctgcgtct gctggaagaa 720
gcaaaagaac gcgatcatcg taaactgggt aaagagctgg aactgtttac caccagtcag 780
caggttggtc agggtctgcc gctgtggctg ccgaaaggtg caaccattcg tcgtattatt 840
gaacgctata tcgtggataa agaagttgca ctgggttacg atcatgttta tacaccggtt 900
ctgggtagcg ttgaactgta taaaaccagc ggtcattggg atcactacaa agaaaatatg 960
tttccgccta tggaaatgga caatgaagaa ctggttctgc gtccgatgaa ttgtccgcat 1020
cacatgatga tctataaaag caaactgcac agctatcgtg aactgccgat tcgtattgca 1080
gaactgggca ccatgcatcg ttatgaaatg agcggtgcac tgaccggtct gcagcgtgtt 1140
cgtggtatga ccctgaatga tgcacatatc tttgttcgtc cggatcagat caaagatgaa 1200
ttcaaacgtg tggtgaacct gatcctggaa gtgtataaag attttggcat cgaagaatac 1260
agcttccgtc tgagttatcg tgatccgcat gataaagaaa aatactatga tgacgatgaa 1320
atgtgggaaa aagcacagcg tatgctgcgt gaagcaatgg atgaattagg tctggattat 1380
tatgaagccg aaggtgaagc agccttttat ggtccgaaac tggatgttca ggttcgtacc 1440
gcactgggaa aagatgaaac cctgagcacc gttcagctgg attttctgct gccggaacgt 1500
ttcgatctga cctatattgg tgaagatggc aaaccgcatc gtccggttgt tattcatcgt 1560
ggtgttgtta gcaccatgga acgttttgtg gcatttctga tcgaagagta taaaggtgca 1620
tttccgacct ggctggcacc ggttcaggtt aaagttattc cggttagtcc ggaagcgcac 1680
ctggattatg catatgatgt tcagcgtacc ctgaaagaac gtggttttcg tgttgaagtt 1740
gatgaacgcg acgaaaaaat cggctataaa atccgtgaag cacagatgca gaaaatcccg 1800
tatatgctgg ttgttggtga taaagaggtt agcgaacgcg cagttaatgt tcgtcgttat 1860
ggtgaaaaag aaagccgtac catgggcctt gatgaattta tggccctgct ggcagatgat 1920
gttcgtgaaa aacgtacccg tctgggcaaa gcacagtaa 1959
<210> 60
<211> 651
<212> PRT
<213> Geobacillus
<400> 60
Met Pro Asp Val Ile Arg Ile Thr Phe Pro Asp Gly Ala Glu Lys Glu
1 5 10 15
Phe Pro Lys Gly Thr Thr Thr Glu Asp Val Ala Ala Ser Ile Ser Pro
20 25 30
Gly Leu Lys Lys Lys Ala Ile Ala Gly Lys Leu Asn Gly Arg Phe Val
35 40 45
Asp Leu Arg Thr Pro Leu His Glu Asp Gly Glu Leu Val Ile Ile Thr
50 55 60
Gln Asp Met Pro Glu Ala Leu Asp Ile Leu Arg His Ser Thr Ala His
65 70 75 80
Leu Met Ala Gln Ala Ile Lys Arg Leu Tyr Gly Asn Val Lys Leu Gly
85 90 95
Val Gly Pro Val Ile Glu Asn Gly Phe Tyr Tyr Asp Ile Asp Met Glu
100 105 110
His Lys Leu Thr Pro Asp Asp Leu Pro Lys Ile Glu Ala Glu Met Arg
115 120 125
Lys Ile Val Lys Glu Asn Leu Asp Ile Val Arg Lys Glu Val Ser Arg
130 135 140
Glu Glu Ala Ile Arg Leu Tyr Glu Glu Ile Gly Asp Glu Leu Lys Leu
145 150 155 160
Glu Leu Ile Ala Asp Ile Pro Glu Gly Glu Pro Ile Ser Ile Tyr Glu
165 170 175
Gln Gly Glu Phe Phe Asp Leu Cys Arg Gly Val His Val Pro Ser Thr
180 185 190
Gly Lys Ile Lys Glu Phe Lys Leu Leu Ser Ile Ser Gly Ala Tyr Trp
195 200 205
Arg Gly Asp Ser Asn Asn Lys Met Leu Gln Arg Ile Tyr Gly Thr Ala
210 215 220
Phe Phe Lys Lys Glu Asp Leu Asp Arg Tyr Leu Arg Leu Leu Glu Glu
225 230 235 240
Ala Lys Glu Arg Asp His Arg Lys Leu Gly Lys Glu Leu Glu Leu Phe
245 250 255
Thr Thr Ser Gln Gln Val Gly Gln Gly Leu Pro Leu Trp Leu Pro Lys
260 265 270
Gly Ala Thr Ile Arg Arg Ile Ile Glu Arg Tyr Ile Val Asp Lys Glu
275 280 285
Val Ala Leu Gly Tyr Asp His Val Tyr Thr Pro Val Leu Gly Ser Val
290 295 300
Glu Leu Tyr Lys Thr Ser Gly His Trp Asp His Tyr Lys Glu Asn Met
305 310 315 320
Phe Pro Pro Met Glu Met Asp Asn Glu Glu Leu Val Leu Arg Pro Met
325 330 335
Asn Cys Pro His His Met Met Ile Tyr Lys Ser Lys Leu His Ser Tyr
340 345 350
Arg Glu Leu Pro Ile Arg Ile Ala Glu Leu Gly Thr Met His Arg Tyr
355 360 365
Glu Met Ser Gly Ala Leu Thr Gly Leu Gln Arg Val Arg Gly Met Thr
370 375 380
Leu Asn Asp Ala His Ile Phe Val Arg Pro Asp Gln Ile Lys Asp Glu
385 390 395 400
Phe Lys Arg Val Val Asn Leu Ile Leu Glu Val Tyr Lys Asp Phe Gly
405 410 415
Ile Glu Glu Tyr Ser Phe Arg Leu Ser Tyr Arg Asp Pro His Asp Lys
420 425 430
Glu Lys Tyr Tyr Asp Asp Asp Glu Met Trp Glu Lys Ala Gln Arg Met
435 440 445
Leu Arg Glu Ala Met Asp Glu Leu Gly Leu Asp Tyr Tyr Glu Ala Glu
450 455 460
Gly Glu Ala Ala Phe Tyr Gly Pro Lys Leu Asp Val Gln Val Arg Ala
465 470 475 480
Leu Gly Lys Asp Glu Thr Leu Ser Thr Val Gln Leu Asp Phe Leu Leu
485 490 495
Pro Glu Arg Phe Asp Leu Thr Tyr Ile Gly Glu Asp Gly Lys Pro His
500 505 510
Arg Pro Val Val Ile His Arg Gly Val Val Ser Thr Met Glu Arg Phe
515 520 525
Val Ala Phe Leu Ile Glu Glu Tyr Lys Gly Ala Phe Pro Thr Trp Leu
530 535 540
Ala Pro Val Gln Val Lys Val Ile Pro Val Ser Pro Glu Ala His Leu
545 550 555 560
Asp Tyr Ala Tyr Asp Val Gln Arg Thr Leu Lys Glu Arg Gly Phe Arg
565 570 575
Val Glu Val Asp Glu Arg Asp Glu Lys Ile Gly Tyr Lys Ile Arg Glu
580 585 590
Ala Gln Met Gln Lys Ile Pro Tyr Met Leu Val Val Gly Asp Lys Glu
595 600 605
Val Ser Glu Arg Ala Val Asn Val Arg Arg Tyr Gly Glu Lys Glu Ser
610 615 620
Arg Thr Met Gly Leu Asp Glu Phe Met Ala Leu Leu Ala Asp Asp Val
625 630 635 640
Arg Glu Lys Arg Thr Arg Leu Gly Lys Ala Gln
645 650
<210> 61
<211> 987
<212> DNA
<213> Geobacillus stearothermophilus
<400> 61
atgaaaacca tctttagcgg tattcagccg agcggtgtta ttaccctggg taactatatt 60
ggtgcactgc gtcagtttat tgaactgcag catgaatata actgctattt ctgcattgtt 120
gatcagcatg caattaccgt ttggcaggat ccgcatgaac tgcgccagaa tattcgtcgt 180
ctggcagcac tgtatctggc agttggtatt gatccgacac aggcaaccct gtttattcag 240
agcgaagttc cggcacatgc acaggcagca tggatgctgc aatgtattgt ttatattggc 300
gaactggaac gcatgaccca gtttaaagaa aaaagcgcag gtaaagaagc agttagcgca 360
ggtctgctga cctatccgcc tctgatggca gccgatattc tgctgtataa caccgatatt 420
gttccggttg gtgatgatca gaaacagcat atcgaactga cccgtgatct ggcagaacgt 480
tttaacaaac gttatggtga gctgtttacc attccggaag cacgtattcc gaaagttggt 540
gcacgtatta tgagcctggt ggatccgacc aaaaaaatga gcaaaagcga tccgaatccg 600
aaagcctata ttacactgct ggatgatgca aaaaccatcg agaaaaaaat caaaagtgcc 660
gtgaccgata gcgaaggcac cattcgttat gataaagaag ccaaaccggg tattagcaac 720
ctgctgaaca tttatagcac cctgagcggt cagagcattg aagaattaga acgtaaatat 780
gaaggcaaag gctacggtgt ttttaaagca gatctggcac aggttgttat tgaaaccctg 840
cgtccgattc aagaacgtta tcatcattgg atggaaagcg aagaactgga tcgtgttctg 900
gatgaaggtg cagaaaaagc aaatcgtgtt gcaagcgaaa tggtgcgtaa aatggaacag 960
gcaatgggtc tgggtcgtcg tcgttaa 987
<210> 62
<211> 328
<212> PRT
<213> Geobacillus stearothermophilus
<400> 62
Met Lys Thr Ile Phe Ser Gly Ile Gln Pro Ser Gly Val Ile Thr Leu
1 5 10 15
Gly Asn Tyr Ile Gly Ala Leu Arg Gln Phe Ile Glu Leu Gln His Glu
20 25 30
Tyr Asn Cys Tyr Phe Cys Ile Val Asp Gln His Ala Ile Thr Val Trp
35 40 45
Gln Asp Pro His Glu Leu Arg Gln Asn Ile Arg Arg Leu Ala Ala Leu
50 55 60
Tyr Leu Ala Val Gly Ile Asp Pro Thr Gln Ala Thr Leu Phe Ile Gln
65 70 75 80
Ser Glu Val Pro Ala His Ala Gln Ala Ala Trp Met Leu Gln Cys Ile
85 90 95
Val Tyr Ile Gly Glu Leu Glu Arg Met Thr Gln Phe Lys Glu Lys Ser
100 105 110
Ala Gly Lys Glu Ala Val Ser Ala Gly Leu Leu Thr Tyr Pro Pro Leu
115 120 125
Met Ala Ala Asp Ile Leu Leu Tyr Asn Thr Asp Ile Val Pro Val Gly
130 135 140
Asp Asp Gln Lys Gln His Ile Glu Leu Thr Arg Asp Leu Ala Glu Arg
145 150 155 160
Phe Asn Lys Arg Tyr Gly Glu Leu Phe Thr Ile Pro Glu Ala Arg Ile
165 170 175
Pro Lys Val Gly Ala Arg Ile Met Ser Leu Val Asp Pro Thr Lys Lys
180 185 190
Met Ser Lys Ser Asp Pro Asn Pro Lys Ala Tyr Ile Thr Leu Leu Asp
195 200 205
Asp Ala Lys Thr Ile Glu Lys Lys Ile Lys Ser Ala Val Thr Asp Ser
210 215 220
Glu Gly Thr Ile Arg Tyr Asp Lys Glu Ala Lys Pro Gly Ile Ser Asn
225 230 235 240
Leu Leu Asn Ile Tyr Ser Thr Leu Ser Gly Gln Ser Ile Glu Glu Leu
245 250 255
Glu Arg Lys Tyr Glu Gly Lys Gly Tyr Gly Val Phe Lys Ala Asp Leu
260 265 270
Ala Gln Val Val Ile Glu Thr Leu Arg Pro Ile Gln Glu Arg Tyr His
275 280 285
His Trp Met Glu Ser Glu Glu Leu Asp Arg Val Leu Asp Glu Gly Ala
290 295 300
Glu Lys Ala Asn Arg Val Ala Ser Glu Met Val Arg Lys Met Glu Gln
305 310 315 320
Ala Met Gly Leu Gly Arg Arg Arg
325
<210> 63
<211> 1260
<212> DNA
<213> Geobacillus stearothermophilus
<400> 63
atggatctgc tggcagaact gcagtggcgt ggtctggtga atcagaccac cgatgaagat 60
ggtctgcgtg aactgctgaa agaagaacgc gttaccctgt attgtggttt tgatccgacc 120
gcagatagcc tgcatattgg taatctggca gcaattctga ccctgcgtcg ttttcagcag 180
gcaggtcatc agccgattgc actggttggt ggtgcaaccg gtctgattgg tgatccgagc 240
ggtaaaaaaa gcgaacgtac cctgaatgca aaagaaaccg ttgaagcatg gtcagcacgt 300
attcaagaac agctgagccg ttttctggat tttgaagcac atggtaatcc ggcaaaaatc 360
aagaacaact atgattggat tggtccgctg gatgttatta cctttctgcg tgatgttggc 420
aaacatttca gcgtgaatta tatgatggcc aaagaaagcg ttcagagccg tattgaaacc 480
ggtattagct ttaccgaatt cagctatatg atgctgcagg cctatgattt tctgcgtctg 540
tatgaaaccg aaggttgtcg tctgcagatt ggtggtagcg atcagtgggg caatattacc 600
gcaggtctgg aactgattcg taaaaccaaa ggtgaagcac gtgcatttgg tctgaccatt 660
ccgctggtta ccaaagcaga tggtacaaaa tttggtaaaa ccgaaagcgg caccatttgg 720
ctggataaag aaaaaaccag tccgtatgag ttctaccagt tttggattaa taccgatgat 780
cgtgatgtga tccgctacct gaaatacttt acatttctga gcaaagaaga gatcgaagcc 840
tttgaacaag aactgcgtga agcaccggaa aaacgtgcag cacagaaagc actggcagaa 900
gaagttacca aactggttca tggtgaagaa gcactgcgtc aggcagttcg tattagcgaa 960
gcactgttta gcggtgatat tggcaacctg accgcagcag aaattgaaca gggttttaaa 1020
gatgttccga gctttgttca tgaaggtggt gatgtgccgc tggtcgaact gctggttagc 1080
gcaggtatta gcccgagcaa acgtcaggca cgtgaagata ttcagaatgg tgccatttat 1140
gtgaatggtg aacgtctgca ggatgttggt gcgattctga cagcagaaca tcgtctggaa 1200
ggtcgtttta ccgttattcg tcgtggcaaa aaaaagtatt acctgattcg ctatgcctaa 1260
<210> 64
<211> 419
<212> PRT
<213> Geobacillus stearothermophilus
<400> 64
Met Asp Leu Leu Ala Glu Leu Gln Trp Arg Gly Leu Val Asn Gln Thr
1 5 10 15
Thr Asp Glu Asp Gly Leu Arg Glu Leu Leu Lys Glu Glu Arg Val Thr
20 25 30
Leu Tyr Cys Gly Phe Asp Pro Thr Ala Asp Ser Leu His Ile Gly Asn
35 40 45
Leu Ala Ala Ile Leu Thr Leu Arg Arg Phe Gln Gln Ala Gly His Gln
50 55 60
Pro Ile Ala Leu Val Gly Gly Ala Thr Gly Leu Ile Gly Asp Pro Ser
65 70 75 80
Gly Lys Lys Ser Glu Arg Thr Leu Asn Ala Lys Glu Thr Val Glu Ala
85 90 95
Trp Ser Ala Arg Ile Gln Glu Gln Leu Ser Arg Phe Leu Asp Phe Glu
100 105 110
Ala His Gly Asn Pro Ala Lys Ile Lys Asn Asn Tyr Asp Trp Ile Gly
115 120 125
Pro Leu Asp Val Ile Thr Phe Leu Arg Asp Val Gly Lys His Phe Ser
130 135 140
Val Asn Tyr Met Met Ala Lys Glu Ser Val Gln Ser Arg Ile Glu Thr
145 150 155 160
Gly Ile Ser Phe Thr Glu Phe Ser Tyr Met Met Leu Gln Ala Tyr Asp
165 170 175
Phe Leu Arg Leu Tyr Glu Thr Glu Gly Cys Arg Leu Gln Ile Gly Gly
180 185 190
Ser Asp Gln Trp Gly Asn Ile Thr Ala Gly Leu Glu Leu Ile Arg Lys
195 200 205
Thr Lys Gly Glu Ala Arg Ala Phe Gly Leu Thr Ile Pro Leu Val Thr
210 215 220
Lys Ala Asp Gly Thr Lys Phe Gly Lys Thr Glu Ser Gly Thr Ile Trp
225 230 235 240
Leu Asp Lys Glu Lys Thr Ser Pro Tyr Glu Phe Tyr Gln Phe Trp Ile
245 250 255
Asn Thr Asp Asp Arg Asp Val Ile Arg Tyr Leu Lys Tyr Phe Thr Phe
260 265 270
Leu Ser Lys Glu Glu Ile Glu Ala Phe Glu Gln Glu Leu Arg Glu Ala
275 280 285
Pro Glu Lys Arg Ala Ala Gln Lys Ala Leu Ala Glu Glu Val Thr Lys
290 295 300
Leu Val His Gly Glu Glu Ala Leu Arg Gln Ala Val Arg Ile Ser Glu
305 310 315 320
Ala Leu Phe Ser Gly Asp Ile Gly Asn Leu Thr Ala Ala Glu Ile Glu
325 330 335
Gln Gly Phe Lys Asp Val Pro Ser Phe Val His Glu Gly Gly Asp Val
340 345 350
Pro Leu Val Glu Leu Leu Val Ser Ala Gly Ile Ser Pro Ser Lys Arg
355 360 365
Gln Ala Arg Glu Asp Ile Gln Asn Gly Ala Ile Tyr Val Asn Gly Glu
370 375 380
Arg Leu Gln Asp Val Gly Ala Ile Leu Thr Ala Glu His Arg Leu Glu
385 390 395 400
Gly Arg Phe Thr Val Ile Arg Arg Gly Lys Lys Lys Tyr Tyr Leu Ile
405 410 415
Arg Tyr Ala
<210> 65
<211> 2643
<212> DNA
<213> Geobacillus
<400> 65
atggcacagc atgaagttag catgcctccg aaatatgatc atcgtgcagt tgaagcaggt 60
cgttatgaat ggtggctgaa aggtaaattc tttgaagcaa ccggtgatcc gaataaacgt 120
ccgtttacca ttgttattcc gcctccgaat gtgaccggta aactgcatct gggtcatgca 180
tgggatacca cactgcagga tattatcacc cgtatgaaac gtatgcaggg ttatgatgtt 240
ctgtggctgc ctggtatgga tcatgcaggt attgcaaccc aggcaaaagt tgaagaaaaa 300
ctgcgtcagc agggtctgag ccgttatgat ctgggtcgtg aaaaatttct ggaagaaacc 360
tggaaatgga aagaagaata cgcaggtcat attcgtagcc agtgggcaaa attaggtctg 420
ggtttagatt atacccgtga acgttttacc ctggatgaag gtctgagcaa agcagttcgt 480
gaagtttttg ttagcctgta tcgtaaaggt ctgatttatc gcggtgagta tatcattaat 540
tgggaccctg ttaccaaaac cgcactgagc gatattgaag tggtttacaa agaagttaaa 600
ggcgcactgt atcatctgcg ttatccgctg gcagatggta gcggttgtat tgaagttgca 660
accacacgtc cggaaaccat gctgggtgat accgcagttg cagttcatcc tgatgatgaa 720
cgttataaac atctgatcgg caaaatggtg aaactgccga ttgttggtcg cgaaattccg 780
attattgcag atgaatatgt ggacatggaa tttggtagtg gtgccgtgaa aattacaccg 840
gcacatgatc cgaacgattt tgaaattggt aatcgccata atctgcctcg tattctggtg 900
atgaatgaag atggcaccat gaatgaaaat gccatgcagt atcaaggtct ggatcgtttt 960
gaatgccgta aacaaattgt tcgcgatctg caagaacagg gtgttctgtt taaaatcgaa 1020
gaacatgtgc atagcgttgg tcatagcgaa cgtagcggtg cagttattga accgtatctg 1080
agcacccagt ggtttgttaa aatgaaaccg ctggccgaag cagcaattaa actgcagcag 1140
accgatggta aagttcagtt tgtgccggaa cgctttgaaa aaacctatct gcattggctg 1200
gaaaacattc gtgattggtg tattagccgt cagctgtggt ggggtcatcg tattccggca 1260
tggtatcata aagaaaccgg tgaaatttat gtggatcacg aaccgcctaa agatatcgaa 1320
aattgggaac aagatccgga tgttctggat acctggttta gcagcgcact gtggccgttt 1380
agcaccatgg gttggcctga tgttgaaagt ccggattata aacgttatta tccgaccgat 1440
gtgctggtta ccggttatga tattatcttt ttttgggtga gccgcatgat ttttcaaggc 1500
ctggaattta ccggcaaacg cccttttaaa gatgttctga ttcatggtct ggtgcgtgat 1560
gcacagggtc gtaaaatgag caaaagctta ggtaatggtg ttgatccgat ggatgtgatt 1620
gatcagtatg gtgcagatgc actgcgttat tttctggcaa ccggtagcag ccctggtcag 1680
gatctgcgtt ttagcaccga aaaagtggaa gcaacgtgga attttgccaa caaaatttgg 1740
aatgcaagcc gttttgcact gatgaacatg ggtggtatga cctatgaaga actggatctg 1800
agcggtgaaa aaacagttgc ggatcattgg attctgaccc gtctgaatga aaccattgat 1860
accgttacca aactggccga aaaatatgaa tttggtgaag ccggtcgtac cctgtataac 1920
tttatttggg atgatctgtg cgattggtat atcgaaatgg caaaactgcc gctgtatggt 1980
gatgatgagg cagcaaaaaa aacaacccgt agcgttctgg catatgtgct ggataatacc 2040
atgcgcctgc tgcatccgtt tatgccgttt attaccgaag aaatttggca gaatctgccg 2100
catgaaggtg aaagcattac cgttgcaccg tggcctcagg ttcgtccgga actgagcaat 2160
gaagaggcag cggaagaaat gcgtatgctg gttgatatta ttcgtgccgt tcgtaatgtt 2220
cgtgccgaag ttaatacccc tccgagcaaa ccgattgcac tgtatatcaa agttaaagac 2280
gaacaggttc gtgcagccct gatgaaaaat cgtgcatatc tggaacgttt ttgcaatccg 2340
agcgaactgc tgattgatac caatgttcct gcaccggata aagcaatgac cgcagtggtg 2400
accggtgcag aactgattat gccgctggaa ggcctgatta acattgaaga agaaattaaa 2460
cgcctggaaa aagaacttga taaatggaac aaagaggtgg aacgcgtcga aaaaaaactg 2520
gcaaatgaag gttttctggc caaagcacca gcgcatgttg tggaagaaga acgtcgtaaa 2580
cgtcaggatt acatggaaaa acgtgaagca gttaaagcac gtctggccga actgaaacgt 2640
taa 2643
<210> 66
<211> 880
<212> PRT
<213> Geobacillus
<400> 66
Met Ala Gln His Glu Val Ser Met Pro Pro Lys Tyr Asp His Arg Ala
1 5 10 15
Val Glu Ala Gly Arg Tyr Glu Trp Trp Leu Lys Gly Lys Phe Phe Glu
20 25 30
Ala Thr Gly Asp Pro Asn Lys Arg Pro Phe Thr Ile Val Ile Pro Pro
35 40 45
Pro Asn Val Thr Gly Lys Leu His Leu Gly His Ala Trp Asp Thr Thr
50 55 60
Leu Gln Asp Ile Ile Thr Arg Met Lys Arg Met Gln Gly Tyr Asp Val
65 70 75 80
Leu Trp Leu Pro Gly Met Asp His Ala Gly Ile Ala Thr Gln Ala Lys
85 90 95
Val Glu Glu Lys Leu Arg Gln Gln Gly Leu Ser Arg Tyr Asp Leu Gly
100 105 110
Arg Glu Lys Phe Leu Glu Glu Thr Trp Lys Trp Lys Glu Glu Tyr Ala
115 120 125
Gly His Ile Arg Ser Gln Trp Ala Lys Leu Gly Leu Gly Leu Asp Tyr
130 135 140
Thr Arg Glu Arg Phe Thr Leu Asp Glu Gly Leu Ser Lys Ala Val Arg
145 150 155 160
Glu Val Phe Val Ser Leu Tyr Arg Lys Gly Leu Ile Tyr Arg Gly Glu
165 170 175
Tyr Ile Ile Asn Trp Asp Pro Val Thr Lys Thr Ala Leu Ser Asp Ile
180 185 190
Glu Val Val Tyr Lys Glu Val Lys Gly Ala Leu Tyr His Leu Arg Tyr
195 200 205
Pro Leu Ala Asp Gly Ser Gly Cys Ile Glu Val Ala Thr Thr Arg Pro
210 215 220
Glu Thr Met Leu Gly Asp Thr Ala Val Ala Val His Pro Asp Asp Glu
225 230 235 240
Arg Tyr Lys His Leu Ile Gly Lys Met Val Lys Leu Pro Ile Val Gly
245 250 255
Arg Glu Ile Pro Ile Ile Ala Asp Glu Tyr Val Asp Met Glu Phe Gly
260 265 270
Ser Gly Ala Val Lys Ile Thr Pro Ala His Asp Pro Asn Asp Phe Glu
275 280 285
Ile Gly Asn Arg His Asn Leu Pro Arg Ile Leu Val Met Asn Glu Asp
290 295 300
Gly Thr Met Asn Glu Asn Ala Met Gln Tyr Gln Gly Leu Asp Arg Phe
305 310 315 320
Glu Cys Arg Lys Gln Ile Val Arg Asp Leu Gln Glu Gln Gly Val Leu
325 330 335
Phe Lys Ile Glu Glu His Val His Ser Val Gly His Ser Glu Arg Ser
340 345 350
Gly Ala Val Ile Glu Pro Tyr Leu Ser Thr Gln Trp Phe Val Lys Met
355 360 365
Lys Pro Leu Ala Glu Ala Ala Ile Lys Leu Gln Gln Thr Asp Gly Lys
370 375 380
Val Gln Phe Val Pro Glu Arg Phe Glu Lys Thr Tyr Leu His Trp Leu
385 390 395 400
Glu Asn Ile Arg Asp Trp Cys Ile Ser Arg Gln Leu Trp Trp Gly His
405 410 415
Arg Ile Pro Ala Trp Tyr His Lys Glu Thr Gly Glu Ile Tyr Val Asp
420 425 430
His Glu Pro Pro Lys Asp Ile Glu Asn Trp Glu Gln Asp Pro Asp Val
435 440 445
Leu Asp Thr Trp Phe Ser Ser Ala Leu Trp Pro Phe Ser Thr Met Gly
450 455 460
Trp Pro Asp Val Glu Ser Pro Asp Tyr Lys Arg Tyr Tyr Pro Thr Asp
465 470 475 480
Val Leu Val Thr Gly Tyr Asp Ile Ile Phe Phe Trp Val Ser Arg Met
485 490 495
Ile Phe Gln Gly Leu Glu Phe Thr Gly Lys Arg Pro Phe Lys Asp Val
500 505 510
Leu Ile His Gly Leu Val Arg Asp Ala Gln Gly Arg Lys Met Ser Lys
515 520 525
Ser Leu Gly Asn Gly Val Asp Pro Met Asp Val Ile Asp Gln Tyr Gly
530 535 540
Ala Asp Ala Leu Arg Tyr Phe Leu Ala Thr Gly Ser Ser Pro Gly Gln
545 550 555 560
Asp Leu Arg Phe Ser Thr Glu Lys Val Glu Ala Thr Trp Asn Phe Ala
565 570 575
Asn Lys Ile Trp Asn Ala Ser Arg Phe Ala Leu Met Asn Met Gly Gly
580 585 590
Met Thr Tyr Glu Glu Leu Asp Leu Ser Gly Glu Lys Thr Val Ala Asp
595 600 605
His Trp Ile Leu Thr Arg Leu Asn Glu Thr Ile Asp Thr Val Thr Lys
610 615 620
Leu Ala Glu Lys Tyr Glu Phe Gly Glu Ala Gly Arg Thr Leu Tyr Asn
625 630 635 640
Phe Ile Trp Asp Asp Leu Cys Asp Trp Tyr Ile Glu Met Ala Lys Leu
645 650 655
Pro Leu Tyr Gly Asp Asp Glu Ala Ala Lys Lys Thr Thr Arg Ser Val
660 665 670
Leu Ala Tyr Val Leu Asp Asn Thr Met Arg Leu Leu His Pro Phe Met
675 680 685
Pro Phe Ile Thr Glu Glu Ile Trp Gln Asn Leu Pro His Glu Gly Glu
690 695 700
Ser Ile Thr Val Ala Pro Trp Pro Gln Val Arg Pro Glu Leu Ser Asn
705 710 715 720
Glu Glu Ala Ala Glu Glu Met Arg Met Leu Val Asp Ile Ile Arg Ala
725 730 735
Val Arg Asn Val Arg Ala Glu Val Asn Thr Pro Pro Ser Lys Pro Ile
740 745 750
Ala Leu Tyr Ile Lys Val Lys Asp Glu Gln Val Arg Ala Ala Leu Met
755 760 765
Lys Asn Arg Ala Tyr Leu Glu Arg Phe Cys Asn Pro Ser Glu Leu Leu
770 775 780
Ile Asp Thr Asn Val Pro Ala Pro Asp Lys Ala Met Thr Ala Val Val
785 790 795 800
Thr Gly Ala Glu Leu Ile Met Pro Leu Glu Gly Leu Ile Asn Ile Glu
805 810 815
Glu Glu Ile Lys Arg Leu Glu Lys Glu Leu Asp Lys Trp Asn Lys Glu
820 825 830
Val Glu Arg Val Glu Lys Lys Leu Ala Asn Glu Gly Phe Leu Ala Lys
835 840 845
Ala Pro Ala His Val Val Glu Glu Glu Arg Arg Lys Arg Gln Asp Tyr
850 855 860
Met Glu Lys Arg Glu Ala Val Lys Ala Arg Leu Ala Glu Leu Lys Arg
865 870 875 880
<210> 67
<211> 960
<212> DNA
<213> Geobacillus stearothermophilus
<400> 67
atgaccaaca ttgtgtttat gggcacaccg gattttgcag ttccgattct gcgtcagctg 60
ctgcatgatg gttatcgtgt tgcagcagtt gttacccagc cggataaacc gaaaggtcgt 120
aaacgtgaac ctgttccgcc tccggttaaa gttgaagcag aacgtcgtgg tattccggtt 180
ctgcagccga ccaaaattcg tgaaccggaa cagtatgaac aggtgctggc atttgcaccg 240
gatctgattg ttaccgcagc atttggtcag attctgccga aagcactgct ggatgcaccg 300
aaatatggtt gcattaatgt tcatgcaagc ctgctgccgg aactgcgtgg tggtgcaccg 360
attcattatg caatttggca gggtaaaacc aaaaccggtg ttaccattat gtatatggtt 420
gaacgtctgg atgccggtga tatgctggca caggttgaag tgccgattgc agaaaccgat 480
accgttggca ccctgcatga taaactgagc gcagcgggtg caaaactgct gagcgaaacc 540
ctgccgctgc tgctggaagg caatattaca ccggttccgc aggatgaaga aaaagcaacc 600
tatgcaccta atattcgtcg tgaacaagaa cgtattgatt ggacccagcc tggtgaagcc 660
atttataacc atattcgtgc ctttcatccg tggcctgtta cctataccac acaggatggt 720
catatttgga aagtttggtg gggtgaaaaa gttcctgcac cgcgtagcgc accgcctggc 780
accattctgg cactggaaga aaatggtatt gttgttgcaa ccggtaatga aaccgcaatt 840
cgtattaccg aactgcagcc tgcaggtaaa aaacgtatgg cagccggtga atttctgcgt 900
ggcgcaggta gccgtctggc agttggtatg aaactgggtg aagatcatga acgtacctaa 960
<210> 68
<211> 319
<212> PRT
<213> Geobacillus stearothermophilus
<400> 68
Met Thr Asn Ile Val Phe Met Gly Thr Pro Asp Phe Ala Val Pro Ile
1 5 10 15
Leu Arg Gln Leu Leu His Asp Gly Tyr Arg Val Ala Ala Val Val Thr
20 25 30
Gln Pro Asp Lys Pro Lys Gly Arg Lys Arg Glu Pro Val Pro Pro Pro
35 40 45
Val Lys Val Glu Ala Glu Arg Arg Gly Ile Pro Val Leu Gln Pro Thr
50 55 60
Lys Ile Arg Glu Pro Glu Gln Tyr Glu Gln Val Leu Ala Phe Ala Pro
65 70 75 80
Asp Leu Ile Val Thr Ala Ala Phe Gly Gln Ile Leu Pro Lys Ala Leu
85 90 95
Leu Asp Ala Pro Lys Tyr Gly Cys Ile Asn Val His Ala Ser Leu Leu
100 105 110
Pro Glu Leu Arg Gly Gly Ala Pro Ile His Tyr Ala Ile Trp Gln Gly
115 120 125
Lys Thr Lys Thr Gly Val Thr Ile Met Tyr Met Val Glu Arg Leu Asp
130 135 140
Ala Gly Asp Met Leu Ala Gln Val Glu Val Pro Ile Ala Glu Thr Asp
145 150 155 160
Thr Val Gly Thr Leu His Asp Lys Leu Ser Ala Ala Gly Ala Lys Leu
165 170 175
Leu Ser Glu Thr Leu Pro Leu Leu Leu Glu Gly Asn Ile Thr Pro Val
180 185 190
Pro Gln Asp Glu Glu Lys Ala Thr Tyr Ala Pro Asn Ile Arg Arg Glu
195 200 205
Gln Glu Arg Ile Asp Trp Thr Gln Pro Gly Glu Ala Ile Tyr Asn His
210 215 220
Ile Arg Ala Phe His Pro Trp Pro Val Thr Tyr Thr Thr Gln Asp Gly
225 230 235 240
His Ile Trp Lys Val Trp Trp Gly Glu Lys Val Pro Ala Pro Arg Ser
245 250 255
Ala Pro Pro Gly Thr Ile Leu Ala Leu Glu Glu Asn Gly Ile Val Val
260 265 270
Ala Thr Gly Asn Glu Thr Ala Ile Arg Ile Thr Glu Leu Gln Pro Ala
275 280 285
Gly Lys Lys Arg Met Ala Ala Gly Glu Phe Leu Arg Gly Ala Gly Ser
290 295 300
Arg Leu Ala Val Gly Met Lys Leu Gly Glu Asp His Glu Arg Thr
305 310 315
<210> 69
<211> 249
<212> DNA
<213> Geobacillus subterraneus
<400> 69
atgttactca ttcgaaggag ggagagccgc tcgatggcaa aagacgatgt aattgaagtg 60
gaaggcaccg tcattgaaac attgccaaat gcgatgtttc gtgtagaatt agaaaatggg 120
cacacagtat tggcccatgt gtccggcaaa atccgtatgc acttcatccg cattttgcct 180
ggcgataaag tgacggtgga gttgtcgccg tatgatttaa cgcgtggacg gattacgtat 240
cgatataaa 249
<210> 70
<211> 83
<212> PRT
<213> Geobacillus subterraneus
<400> 70
Met Leu Leu Ile Arg Arg Arg Glu Ser Arg Ser Met Ala Lys Asp Asp
1 5 10 15
Val Ile Glu Val Glu Gly Thr Val Ile Glu Thr Leu Pro Asn Ala Met
20 25 30
Phe Arg Val Glu Leu Glu Asn Gly His Thr Val Leu Ala His Val Ser
35 40 45
Gly Lys Ile Arg Met His Phe Ile Arg Ile Leu Pro Gly Asp Lys Val
50 55 60
Thr Val Glu Leu Ser Pro Tyr Asp Leu Thr Arg Gly Arg Ile Thr Tyr
65 70 75 80
Arg Tyr Lys
<210> 71
<211> 2352
<212> DNA
<213> Geobacillus subterraneus
<400> 71
atggtgtccc gctttgcaaa gtgccggacc ggtatacgct cggcggcgcg atcggcaaag 60
acgcccgcgt cgttgtcgcc gtcaccgacg aagggttcgc gcgccaattg caaacgatgc 120
tcgactgatc tttatggggg tgaatgtatg tcgaaaatgc gtgtgtacga atacgccaaa 180
aaacataatg tgccaagcaa ggacgttatt cataaattga aagaaatgaa tattgaagtg 240
aacaaccata tgactatgct cgaagccgat gtcgtcgaaa agctcgatca tcaataccgc 300
gtgaactcag agaaaaaagc ggaaaagaaa acggagaaac cgaagcggcc gacgccggcg 360
aaagccgccg attttgccga cgaggaaatg tttgaggaca agaaagaaac ggcaaagacg 420
aagccggcga agaaaaaggg agcagtgaaa ggaaaggaaa cgaaaaaaac agaagcacag 480
cagcaagaaa agaaactgtt ccaagcggcg aagaaaaaag gaaaaggacc gatgaaaggc 540
aaaaaacaag ctgccccagc ctcaaagcag gcgcagcagc cggcgaaaaa agaaaaagag 600
ctcccgaaaa aaattacgtt cgaaggttcg ctcacggtag ccgaattggc gaaaaaactt 660
ggccgcgagc cgtcggaaat cattaaaaaa ctgtttatgc tcggcgtcat ggcgacgatt 720
aaccaagatt tagacaaaga tgcgatcgag ctcatttgct ctgattacgg agttgaagtc 780
gaagaaaaag tgacgatcga tgaaacgaat tttgaaacga tcgaaattgt cgatgcaccg 840
gaagatttgg tggaacggcc gccggtcgtc acgattatgg ggcacgttga ccacgggaaa 900
acaacgctgc ttgacgcaat ccgccactcg aaagtgaccg agcaagaggc gggcggtatt 960
acacagcata tcggtgctta tcaagtcacg gtcaacggca agaaaattac gttcctcgat 1020
acgccggggc atgaagcgtt tacgacgatg cgggcgcgcg gtgcgcaagt gacggatatc 1080
gtcatccttg ttgttgctgc tgatgatggg gtcatgccgc agacggtcga ggcgattaac 1140
cacgccaaag cggcgaacgt accgattatc gtcgccatta acaaaatgga taagccggaa 1200
gcaaacccgg atcgcgttat gcaagagttg atggagtaca acctcgttcc ggaagaatgg 1260
ggtggcgata cgattttctg caagctgtcg gcgaaaaccc aagacggtat tgaccatctg 1320
ttggaaatga ttttgcttgt cagcgaaatg gaagaactaa aagcgaaccc gaaccgccgc 1380
gcgctcggta cggtgatcga agcgaagctc gataaagggc gcggtccggt agcgacgttg 1440
ctcgtccaag ccggtacgct aaaagtcggt gatccgattg ttgtcggaac aacgtacgga 1500
cgcgtgcgcg cgatggtcaa tgacagcggt cggcgtgtca aagaagcggg tccgtcgatg 1560
ccggtcgaaa tcacagggct tcatgatgtg ccgcaagccg gggaccgctt tatggtattt 1620
gaagatgaga agaaagcgcg acaaatcgga gaagcgcggg cacagcggca gctgcaagag 1680
cagcggagcg tgaaaacgcg cgtcagcttg gacgatttgt ttgaacaaat taagcaaggt 1740
gaaatgaaag agctgaactt gatcgttaag gccgacgtcc aaggatcggt cgaagcgctt 1800
gtcgccgcct tgcaaaaaat cgatatcgaa ggcgtgcgtg tgaaaattat ccacgcggcg 1860
gtcggcgcca ttacggagtc agacatcttg ttggcaacga cctcgaacgc gatcgtcatc 1920
ggttttaacg tccgtccgga caccaatgcg aagcgggctg ccgaatcaga aaacgtcgac 1980
atccgcctcc accgcattat ttacaatgtc atcgaagaaa ttgaagcggc gatgaaaggg 2040
atgctcgacc cagaatatga agaaaaagtg atcggtcagg cggaagtgcg gcaaacgttc 2100
aaagtgtcga aagtcggcac gatcgccggg tgctacgtca ccgacggcaa aattacccgc 2160
gacagcaaag tgcgccttat ccgtcaaggc atcgtcgtgt acgaaggcga aatcgactcg 2220
ctcaaacggt ataaagatga tgtgcgtgag gtggcgcaag gatacgaatg cggcgtgacc 2280
atcaaaaact tcaacgatat taaagaaggg gacgtcatcg aggcgtacat catgcaggaa 2340
gtggctcgcg ca 2352
<210> 72
<211> 784
<212> PRT
<213> Geobacillus subterraneus
<400> 72
Met Val Ser Arg Phe Ala Lys Cys Arg Thr Gly Ile Arg Ser Ala Ala
1 5 10 15
Arg Ser Ala Lys Thr Pro Ala Ser Leu Ser Pro Ser Pro Thr Lys Gly
20 25 30
Ser Arg Ala Asn Cys Lys Arg Cys Ser Thr Asp Leu Tyr Gly Gly Glu
35 40 45
Cys Met Ser Lys Met Arg Val Tyr Glu Tyr Ala Lys Lys His Asn Val
50 55 60
Pro Ser Lys Asp Val Ile His Lys Leu Lys Glu Met Asn Ile Glu Val
65 70 75 80
Asn Asn His Met Thr Met Leu Glu Ala Asp Val Val Glu Lys Leu Asp
85 90 95
His Gln Tyr Arg Val Asn Ser Glu Lys Lys Ala Glu Lys Lys Thr Glu
100 105 110
Lys Pro Lys Arg Pro Thr Pro Ala Lys Ala Ala Asp Phe Ala Asp Glu
115 120 125
Glu Met Phe Glu Asp Lys Lys Glu Thr Ala Lys Thr Lys Pro Ala Lys
130 135 140
Lys Lys Gly Ala Val Lys Gly Lys Glu Thr Lys Lys Thr Glu Ala Gln
145 150 155 160
Gln Gln Glu Lys Lys Leu Phe Gln Ala Ala Lys Lys Lys Gly Lys Gly
165 170 175
Pro Met Lys Gly Lys Lys Gln Ala Ala Pro Ala Ser Lys Gln Ala Gln
180 185 190
Gln Pro Ala Lys Lys Glu Lys Glu Leu Pro Lys Lys Ile Thr Phe Glu
195 200 205
Gly Ser Leu Thr Val Ala Glu Leu Ala Lys Lys Leu Gly Arg Glu Pro
210 215 220
Ser Glu Ile Ile Lys Lys Leu Phe Met Leu Gly Val Met Ala Thr Ile
225 230 235 240
Asn Gln Asp Leu Asp Lys Asp Ala Ile Glu Leu Ile Cys Ser Asp Tyr
245 250 255
Gly Val Glu Val Glu Glu Lys Val Thr Ile Asp Glu Thr Asn Phe Glu
260 265 270
Thr Ile Glu Ile Val Asp Ala Pro Glu Asp Leu Val Glu Arg Pro Pro
275 280 285
Val Val Thr Ile Met Gly His Val Asp His Gly Lys Thr Thr Leu Leu
290 295 300
Asp Ala Ile Arg His Ser Lys Val Thr Glu Gln Glu Ala Gly Gly Ile
305 310 315 320
Thr Gln His Ile Gly Ala Tyr Gln Val Thr Val Asn Gly Lys Lys Ile
325 330 335
Thr Phe Leu Asp Thr Pro Gly His Glu Ala Phe Thr Thr Met Arg Ala
340 345 350
Arg Gly Ala Gln Val Thr Asp Ile Val Ile Leu Val Val Ala Ala Asp
355 360 365
Asp Gly Val Met Pro Gln Thr Val Glu Ala Ile Asn His Ala Lys Ala
370 375 380
Ala Asn Val Pro Ile Ile Val Ala Ile Asn Lys Met Asp Lys Pro Glu
385 390 395 400
Ala Asn Pro Asp Arg Val Met Gln Glu Leu Met Glu Tyr Asn Leu Val
405 410 415
Pro Glu Glu Trp Gly Gly Asp Thr Ile Phe Cys Lys Leu Ser Ala Lys
420 425 430
Thr Gln Asp Gly Ile Asp His Leu Leu Glu Met Ile Leu Leu Val Ser
435 440 445
Glu Met Glu Glu Leu Lys Ala Asn Pro Asn Arg Arg Ala Leu Gly Thr
450 455 460
Val Ile Glu Ala Lys Leu Asp Lys Gly Arg Gly Pro Val Ala Thr Leu
465 470 475 480
Leu Val Gln Ala Gly Thr Leu Lys Val Gly Asp Pro Ile Val Val Gly
485 490 495
Thr Thr Tyr Gly Arg Val Arg Ala Met Val Asn Asp Ser Gly Arg Arg
500 505 510
Val Lys Glu Ala Gly Pro Ser Met Pro Val Glu Ile Thr Gly Leu His
515 520 525
Asp Val Pro Gln Ala Gly Asp Arg Phe Met Val Phe Glu Asp Glu Lys
530 535 540
Lys Ala Arg Gln Ile Gly Glu Ala Arg Ala Gln Arg Gln Leu Gln Glu
545 550 555 560
Gln Arg Ser Val Lys Thr Arg Val Ser Leu Asp Asp Leu Phe Glu Gln
565 570 575
Ile Lys Gln Gly Glu Met Lys Glu Leu Asn Leu Ile Val Lys Ala Asp
580 585 590
Val Gln Gly Ser Val Glu Ala Leu Val Ala Ala Leu Gln Lys Ile Asp
595 600 605
Ile Glu Gly Val Arg Val Lys Ile Ile His Ala Ala Val Gly Ala Ile
610 615 620
Thr Glu Ser Asp Ile Leu Leu Ala Thr Thr Ser Asn Ala Ile Val Ile
625 630 635 640
Gly Phe Asn Val Arg Pro Asp Thr Asn Ala Lys Arg Ala Ala Glu Ser
645 650 655
Glu Asn Val Asp Ile Arg Leu His Arg Ile Ile Tyr Asn Val Ile Glu
660 665 670
Glu Ile Glu Ala Ala Met Lys Gly Met Leu Asp Pro Glu Tyr Glu Glu
675 680 685
Lys Val Ile Gly Gln Ala Glu Val Arg Gln Thr Phe Lys Val Ser Lys
690 695 700
Val Gly Thr Ile Ala Gly Cys Tyr Val Thr Asp Gly Lys Ile Thr Arg
705 710 715 720
Asp Ser Lys Val Arg Leu Ile Arg Gln Gly Ile Val Val Tyr Glu Gly
725 730 735
Glu Ile Asp Ser Leu Lys Arg Tyr Lys Asp Asp Val Arg Glu Val Ala
740 745 750
Gln Gly Tyr Glu Cys Gly Val Thr Ile Lys Asn Phe Asn Asp Ile Lys
755 760 765
Glu Gly Asp Val Ile Glu Ala Tyr Ile Met Gln Glu Val Ala Arg Ala
770 775 780
<210> 73
<211> 339
<212> DNA
<213> Geobacillus subterraneus
<400> 73
atggactacg gcaaattccg ctttgagcag caaaagaaag aaaaagaagc gcgcaaaaag 60
caaaaggtga tcaacattaa agaggtgcgc ctcagcccga caattgagga acacgacttt 120
aatacgaaac tacgcaatgc gcgcaagttt ttagaaaaag gcgataaagt gaaggcgacg 180
atccgcttta aagggcgggc gatcacccat aaagaaatcg ggcagcgcgt ccttgaccgc 240
ttctcggaag catgcgctga tatcgcggtc gtcgaaacgg cgccgaaatt ggaagggcgc 300
aacatgtttt tagtgctggc accgaaaaat gacaacaag 339
<210> 74
<211> 113
<212> PRT
<213> Geobacillus subterraneus
<400> 74
Met Asp Tyr Gly Lys Phe Arg Phe Glu Gln Gln Lys Lys Glu Lys Glu
1 5 10 15
Ala Arg Lys Lys Gln Lys Val Ile Asn Ile Lys Glu Val Arg Leu Ser
20 25 30
Pro Thr Ile Glu Glu His Asp Phe Asn Thr Lys Leu Arg Asn Ala Arg
35 40 45
Lys Phe Leu Glu Lys Gly Asp Lys Val Lys Ala Thr Ile Arg Phe Lys
50 55 60
Gly Arg Ala Ile Thr His Lys Glu Ile Gly Gln Arg Val Leu Asp Arg
65 70 75 80
Phe Ser Glu Ala Cys Ala Asp Ile Ala Val Val Glu Thr Ala Pro Lys
85 90 95
Leu Glu Gly Arg Asn Met Phe Leu Val Leu Ala Pro Lys Asn Asp Asn
100 105 110
Lys
<210> 75
<211> 2076
<212> DNA
<213> Geobacillus subterraneus
<400> 75
atggcaagag agttctcctt agaaaacact cgtaacatag gaatcatggc gcacattgac 60
gccggaaaaa cgacgacgac ggaacgaatc ctgttctaca caggccgcgt tcataaaatc 120
ggggaaacgc atgaaggctc agctacgatg gactggatgg aacaagagca agagcgcggg 180
attacgatta cgtcggcggc gacaacggcg caatggaaag gccatcgcat caacatcatc 240
gacacgccag ggcacgtcga cttcacggtt gaggttgaac gttcgttgcg cgtgttggac 300
ggagccatta cagttcttga cgcccaatct ggtgtagaac cgcaaacgga aacagtttgg 360
cgtcaagcga ctacatatgg tgttccgcgg attgtattcg tcaacaaaat ggacaaaatc 420
ggtgcggact tcttgtatgc ggtaaaaacg ctccatgacc gcttacaagc gaatgcctac 480
ccggtgcagt tgccgatcgg cgctgaagac caattcaccg gcattattga cctcgtggaa 540
atgtgtgcat accattacca cgacgacctt ggcaaaaaca tcgaacgcat cgaaattccg 600
gaagactacc gcgatttagc ggaagaatat catggcaagc tcattgaggc tgttgcggaa 660
ctcgatgaag agctgatgat gaaatattta gaaggagaag aaattacgaa agaagagctg 720
aaagccgcaa tccgtaaggc gacgatcaac gttgaattct atccagtctt ctgcggttca 780
gcttttaaaa acaaaggtgt tcagctgctt cttgacgggg ttgtcgacta cttgccgtct 840
ccgttagata tcccggcgat tcgcggtatc attccggata cggaagaaga agtggctcgc 900
gaagcacgcg atgacgctcc gttctccgcg ttggcattca aaattatgac tgacccgtac 960
gttgggaagt tgacgttctt ccgcgtctac tccggaacgc ttgattccgg ttcttacgtc 1020
atgaactcaa cgaaacggaa gcgtgaacgg atcggtcgct tgctgcaaat gcatgcgaac 1080
caccgtcaag aaatttcgac agtctatgcc ggtgatattg cggcagcagt aggtttaaaa 1140
gaaacaacga ccggcgatac tctatgtgat gagaaaaatc ttgtcatctt agagtcgatg 1200
caattcccag agccggttat ctcggtggcg atcgaaccga aatcgaaagc cgaccaagat 1260
aagatgggtc aagcattgca aaaactgcaa gaggaagacc cgacattccg tgcgcatacc 1320
gatccggaaa caggacaaac gatcatttcc gggatgggcg agctgcactt ggacattatc 1380
gtcgaccgga tgcgtcgcga attcaaagtc gaggcgaacg ttggtgcacc gcaagttgct 1440
taccgtgaaa cgttccgtca atcggctcaa gtcgaaggga aatttattcg ccagtccggt 1500
ggtcgtggtc agtacggtca cgtttggatc gaattcacac cgaacgaacg cggtaaaggc 1560
tttgaatttg aaaatgcgat cgtcggtggg gtcgttccga aagagtacgt gccggctgtt 1620
caagctggat tggaagaagc gatgcaaaac ggtgtcttag ctggctaccc ggttgttgac 1680
atcaaagcga aactgtttga tggatcgtac catgatgtcg actcgagtga gatggcgttc 1740
aaaattgctg cttcgatggc gttgaaaaac gcggcagcga agtgtgaacc ggttctgctt 1800
gaaccgatca tgaaagtaga agtcgtcatc cctgaagaat acctcggcga cattatgggt 1860
gacatcacat cccgccgcgg tcgcgtcgaa gggatggaag cgcgcggaaa cgcccaagtt 1920
gttcgtgcaa tggtgccgct ggccgaaatg ttcggttatg caacatcgct ccgttcgaac 1980
acgcaagggc gtggaacgtt ctcgatggta tttgaccatt acgaagaagt tccgaaaaac 2040
atcgccgatg aaattatcaa aaaaaataaa ggcgaa 2076
<210> 76
<211> 692
<212> PRT
<213> Geobacillus subterraneus
<400> 76
Met Ala Arg Glu Phe Ser Leu Glu Asn Thr Arg Asn Ile Gly Ile Met
1 5 10 15
Ala His Ile Asp Ala Gly Lys Thr Thr Thr Thr Glu Arg Ile Leu Phe
20 25 30
Tyr Thr Gly Arg Val His Lys Ile Gly Glu Thr His Glu Gly Ser Ala
35 40 45
Thr Met Asp Trp Met Glu Gln Glu Gln Glu Arg Gly Ile Thr Ile Thr
50 55 60
Ser Ala Ala Thr Thr Ala Gln Trp Lys Gly His Arg Ile Asn Ile Ile
65 70 75 80
Asp Thr Pro Gly His Val Asp Phe Thr Val Glu Val Glu Arg Ser Leu
85 90 95
Arg Val Leu Asp Gly Ala Ile Thr Val Leu Asp Ala Gln Ser Gly Val
100 105 110
Glu Pro Gln Thr Glu Thr Val Trp Arg Gln Ala Thr Thr Tyr Gly Val
115 120 125
Pro Arg Ile Val Phe Val Asn Lys Met Asp Lys Ile Gly Ala Asp Phe
130 135 140
Leu Tyr Ala Val Lys Thr Leu His Asp Arg Leu Gln Ala Asn Ala Tyr
145 150 155 160
Pro Val Gln Leu Pro Ile Gly Ala Glu Asp Gln Phe Thr Gly Ile Ile
165 170 175
Asp Leu Val Glu Met Cys Ala Tyr His Tyr His Asp Asp Leu Gly Lys
180 185 190
Asn Ile Glu Arg Ile Glu Ile Pro Glu Asp Tyr Arg Asp Leu Ala Glu
195 200 205
Glu Tyr His Gly Lys Leu Ile Glu Ala Val Ala Glu Leu Asp Glu Glu
210 215 220
Leu Met Met Lys Tyr Leu Glu Gly Glu Glu Ile Thr Lys Glu Glu Leu
225 230 235 240
Lys Ala Ala Ile Arg Lys Ala Thr Ile Asn Val Glu Phe Tyr Pro Val
245 250 255
Phe Cys Gly Ser Ala Phe Lys Asn Lys Gly Val Gln Leu Leu Leu Asp
260 265 270
Gly Val Val Asp Tyr Leu Pro Ser Pro Leu Asp Ile Pro Ala Ile Arg
275 280 285
Gly Ile Ile Pro Asp Thr Glu Glu Glu Val Ala Arg Glu Ala Arg Asp
290 295 300
Asp Ala Pro Phe Ser Ala Leu Ala Phe Lys Ile Met Thr Asp Pro Tyr
305 310 315 320
Val Gly Lys Leu Thr Phe Phe Arg Val Tyr Ser Gly Thr Leu Asp Ser
325 330 335
Gly Ser Tyr Val Met Asn Ser Thr Lys Arg Lys Arg Glu Arg Ile Gly
340 345 350
Arg Leu Leu Gln Met His Ala Asn His Arg Gln Glu Ile Ser Thr Val
355 360 365
Tyr Ala Gly Asp Ile Ala Ala Ala Val Gly Leu Lys Glu Thr Thr Thr
370 375 380
Gly Asp Thr Leu Cys Asp Glu Lys Asn Leu Val Ile Leu Glu Ser Met
385 390 395 400
Gln Phe Pro Glu Pro Val Ile Ser Val Ala Ile Glu Pro Lys Ser Lys
405 410 415
Ala Asp Gln Asp Lys Met Gly Gln Ala Leu Gln Lys Leu Gln Glu Glu
420 425 430
Asp Pro Thr Phe Arg Ala His Thr Asp Pro Glu Thr Gly Gln Thr Ile
435 440 445
Ile Ser Gly Met Gly Glu Leu His Leu Asp Ile Ile Val Asp Arg Met
450 455 460
Arg Arg Glu Phe Lys Val Glu Ala Asn Val Gly Ala Pro Gln Val Ala
465 470 475 480
Tyr Arg Glu Thr Phe Arg Gln Ser Ala Gln Val Glu Gly Lys Phe Ile
485 490 495
Arg Gln Ser Gly Gly Arg Gly Gln Tyr Gly His Val Trp Ile Glu Phe
500 505 510
Thr Pro Asn Glu Arg Gly Lys Gly Phe Glu Phe Glu Asn Ala Ile Val
515 520 525
Gly Gly Val Val Pro Lys Glu Tyr Val Pro Ala Val Gln Ala Gly Leu
530 535 540
Glu Glu Ala Met Gln Asn Gly Val Leu Ala Gly Tyr Pro Val Val Asp
545 550 555 560
Ile Lys Ala Lys Leu Phe Asp Gly Ser Tyr His Asp Val Asp Ser Ser
565 570 575
Glu Met Ala Phe Lys Ile Ala Ala Ser Met Ala Leu Lys Asn Ala Ala
580 585 590
Ala Lys Cys Glu Pro Val Leu Leu Glu Pro Ile Met Lys Val Glu Val
595 600 605
Val Ile Pro Glu Glu Tyr Leu Gly Asp Ile Met Gly Asp Ile Thr Ser
610 615 620
Arg Arg Gly Arg Val Glu Gly Met Glu Ala Arg Gly Asn Ala Gln Val
625 630 635 640
Val Arg Ala Met Val Pro Leu Ala Glu Met Phe Gly Tyr Ala Thr Ser
645 650 655
Leu Arg Ser Asn Thr Gln Gly Arg Gly Thr Phe Ser Met Val Phe Asp
660 665 670
His Tyr Glu Glu Val Pro Lys Asn Ile Ala Asp Glu Ile Ile Lys Lys
675 680 685
Asn Lys Gly Glu
690
<210> 77
<211> 1185
<212> DNA
<213> Geobacillus subterraneus
<400> 77
atggctaaag cgaaatttga gcgtacgaaa ccgcacgtca acattggcac gatcggccac 60
gttgaccatg ggaaaacgac gttgacagct gcgatcacga cagttcttgc gaaacaaggt 120
aaagcagaag cgagagcgta cgaccaaatc gacgctgctc cggaagagcg tgaacgcgga 180
atcacgattt cgacggctca cgttgagtat gaaacagaaa accgtcacta tgcgcacgtt 240
gactgcccgg gccacgctga ctacgtgaaa aacatgatca cgggcgcagc gcaaatggac 300
ggcgcgatcc ttgttgtatc ggctgctgac ggtccgatgc cgcaaactcg cgaacacatt 360
cttctttccc gccaagtcgg tgttccgtac atcgttgttt tcttgaacaa atgcgacatg 420
gtggacgacg aagaattgct tgaactcgtt gaaatggaag ttcgcgatct tctttctgaa 480
tatgacttcc cgggcgacga agtgccggtt atcaaaggtt cggcattaaa agcgctcgaa 540
ggcgatgcac aatgggaaga aaaaatcgtt gaactgatga acgcggttga cgagtacatc 600
ccaactccgc aacgtgaagt agacaaaccg ttcatgatgc cggttgagga cgtcttctcg 660
atcacgggtc gtggtacggt tgcaacgggc cgtgttgagc gcggtacgtt aaaagttggt 720
gacccggttg aaatcatcgg tctttcggac gagccgaaat cgacgactgt tacgggtgta 780
gaaatgttcc gtaagcttct cgaccaagca gaagctggtg acaacatcgg tgcgcttctc 840
cgcggtgtat cgcgtgacga agttgagcgc ggtcaagtat tggcgaaacc gggctcgatc 900
acgccacaca cgaaatttaa agcacaagtt tacgttctga cgaaagaaga aggcggacgc 960
catactccgt tcttctcgaa ctaccgtccg caattctact tccgtacaac ggacgtaacg 1020
ggcatcatca cgcttccaga aggcgttgaa atggttatgc ctggcgacaa cgttgaaatg 1080
acggttgaac tgatcgctcc gatcgcgatc gaagaaggta cgaaattctc gatccgtgaa 1140
ggcggccgca cggttggtgc tggttccgta tcggaaatca ttgag 1185
<210> 78
<211> 395
<212> PRT
<213> Geobacillus subterraneus
<400> 78
Met Ala Lys Ala Lys Phe Glu Arg Thr Lys Pro His Val Asn Ile Gly
1 5 10 15
Thr Ile Gly His Val Asp His Gly Lys Thr Thr Leu Thr Ala Ala Ile
20 25 30
Thr Thr Val Leu Ala Lys Gln Gly Lys Ala Glu Ala Arg Ala Tyr Asp
35 40 45
Gln Ile Asp Ala Ala Pro Glu Glu Arg Glu Arg Gly Ile Thr Ile Ser
50 55 60
Thr Ala His Val Glu Tyr Glu Thr Glu Asn Arg His Tyr Ala His Val
65 70 75 80
Asp Cys Pro Gly His Ala Asp Tyr Val Lys Asn Met Ile Thr Gly Ala
85 90 95
Ala Gln Met Asp Gly Ala Ile Leu Val Val Ser Ala Ala Asp Gly Pro
100 105 110
Met Pro Gln Thr Arg Glu His Ile Leu Leu Ser Arg Gln Val Gly Val
115 120 125
Pro Tyr Ile Val Val Phe Leu Asn Lys Cys Asp Met Val Asp Asp Glu
130 135 140
Glu Leu Leu Glu Leu Val Glu Met Glu Val Arg Asp Leu Leu Ser Glu
145 150 155 160
Tyr Asp Phe Pro Gly Asp Glu Val Pro Val Ile Lys Gly Ser Ala Leu
165 170 175
Lys Ala Leu Glu Gly Asp Ala Gln Trp Glu Glu Lys Ile Val Glu Leu
180 185 190
Met Asn Ala Val Asp Glu Tyr Ile Pro Thr Pro Gln Arg Glu Val Asp
195 200 205
Lys Pro Phe Met Met Pro Val Glu Asp Val Phe Ser Ile Thr Gly Arg
210 215 220
Gly Thr Val Ala Thr Gly Arg Val Glu Arg Gly Thr Leu Lys Val Gly
225 230 235 240
Asp Pro Val Glu Ile Ile Gly Leu Ser Asp Glu Pro Lys Ser Thr Thr
245 250 255
Val Thr Gly Val Glu Met Phe Arg Lys Leu Leu Asp Gln Ala Glu Ala
260 265 270
Gly Asp Asn Ile Gly Ala Leu Leu Arg Gly Val Ser Arg Asp Glu Val
275 280 285
Glu Arg Gly Gln Val Leu Ala Lys Pro Gly Ser Ile Thr Pro His Thr
290 295 300
Lys Phe Lys Ala Gln Val Tyr Val Leu Thr Lys Glu Glu Gly Gly Arg
305 310 315 320
His Thr Pro Phe Phe Ser Asn Tyr Arg Pro Gln Phe Tyr Phe Arg Thr
325 330 335
Thr Asp Val Thr Gly Ile Ile Thr Leu Pro Glu Gly Val Glu Met Val
340 345 350
Met Pro Gly Asp Asn Val Glu Met Thr Val Glu Leu Ile Ala Pro Ile
355 360 365
Ala Ile Glu Glu Gly Thr Lys Phe Ser Ile Arg Glu Gly Gly Arg Thr
370 375 380
Val Gly Ala Gly Ser Val Ser Glu Ile Ile Glu
385 390 395
<210> 79
<211> 882
<212> DNA
<213> Geobacillus subterraneus
<400> 79
atggcgatta cagcacaaat ggtaaaagag ctgcgcgaaa aaacgggcgc aggcatgatg 60
gactgcaaaa aagcgctcac cgaaacgaac ggtgacatgg aaaaagcgat cgactggctg 120
cgtgaaaaag gaattgctaa agcagcgaaa aaagcagatc gcatcgcagc ggaaggaatg 180
acatacatcg cgacggaagg caatgcggct gtcattttgg aagtaaactc ggaaacggac 240
ttcgttgcca aaaacgaagc gttccaaacg ctcgttaagg agctggctgc acatctgctg 300
aaacaaaagc cagccacgct tgatgaagcg ctcggacaaa cgatgagcag tggttccact 360
gttcaagatt acattaacga agcagttgct aaaatcggtg aaaaaattac gctccgccgc 420
tttgctgttg tcaacaaagc ggatgatgaa acgtttggcg cgtacttgca catgggcggg 480
cgcatcggcg tattaacatt attagccggc aacgcaactg aagaggtcgc taaagatgtg 540
gcgatgcata ttgctgcgct ccatccgaaa tacgtttcgc gcgatgaagt gccgcaagaa 600
gagattgcgc gcgaacgtga agtgttgaaa caacaagcgt tgaacgaagg taagccggaa 660
aacatcgttg aaaaaatggt tgaaggccgt ctgaaaaagt tttacgaaga tgtttgcctg 720
cttgagcaag cgttcgtgaa aaacccggat gtgacggtac gccaatacgt cgaatcgagc 780
ggagcaaccg tgaagcagtt catccgctac gaagttggtg aagggctcga aaaacgtcaa 840
gataatttcg ctgaagaagt catgagccaa gtaagaaaac aa 882
<210> 80
<211> 294
<212> PRT
<213> Geobacillus subterraneus
<400> 80
Met Ala Ile Thr Ala Gln Met Val Lys Glu Leu Arg Glu Lys Thr Gly
1 5 10 15
Ala Gly Met Met Asp Cys Lys Lys Ala Leu Thr Glu Thr Asn Gly Asp
20 25 30
Met Glu Lys Ala Ile Asp Trp Leu Arg Glu Lys Gly Ile Ala Lys Ala
35 40 45
Ala Lys Lys Ala Asp Arg Ile Ala Ala Glu Gly Met Thr Tyr Ile Ala
50 55 60
Thr Glu Gly Asn Ala Ala Val Ile Leu Glu Val Asn Ser Glu Thr Asp
65 70 75 80
Phe Val Ala Lys Asn Glu Ala Phe Gln Thr Leu Val Lys Glu Leu Ala
85 90 95
Ala His Leu Leu Lys Gln Lys Pro Ala Thr Leu Asp Glu Ala Leu Gly
100 105 110
Gln Thr Met Ser Ser Gly Ser Thr Val Gln Asp Tyr Ile Asn Glu Ala
115 120 125
Val Ala Lys Ile Gly Glu Lys Ile Thr Leu Arg Arg Phe Ala Val Val
130 135 140
Asn Lys Ala Asp Asp Glu Thr Phe Gly Ala Tyr Leu His Met Gly Gly
145 150 155 160
Arg Ile Gly Val Leu Thr Leu Leu Ala Gly Asn Ala Thr Glu Glu Val
165 170 175
Ala Lys Asp Val Ala Met His Ile Ala Ala Leu His Pro Lys Tyr Val
180 185 190
Ser Arg Asp Glu Val Pro Gln Glu Glu Ile Ala Arg Glu Arg Glu Val
195 200 205
Leu Lys Gln Gln Ala Leu Asn Glu Gly Lys Pro Glu Asn Ile Val Glu
210 215 220
Lys Met Val Glu Gly Arg Leu Lys Lys Phe Tyr Glu Asp Val Cys Leu
225 230 235 240
Leu Glu Gln Ala Phe Val Lys Asn Pro Asp Val Thr Val Arg Gln Tyr
245 250 255
Val Glu Ser Ser Gly Ala Thr Val Lys Gln Phe Ile Arg Tyr Glu Val
260 265 270
Gly Glu Gly Leu Glu Lys Arg Gln Asp Asn Phe Ala Glu Glu Val Met
275 280 285
Ser Gln Val Arg Lys Gln
290
<210> 81
<211> 1827
<212> DNA
<213> Geobacillus subterraneus
<400> 81
atgaaccggg aagaacggtt gaaacggcag gaacggattc gcaacttttc gattatcgct 60
cacattgacc acggaaaatc gacgcttgcg gaccgcattt tagaaaaaac aggtgcgctg 120
tcggagcgcg agttgcgcga gcagacgctc gatatgatgg agctcgagcg cgagcgcggc 180
atcacgatca aattgaatgc ggtccagttg acatataaag cgaaaaacgg ggaagagtat 240
attttccatt tgatcgatac gccgggccac gtcgatttta cgtatgaagt gtcgcgcagc 300
ttggctgctt gcgaaggagc gatcttagtc gtcgatgcgg cgcaaggcat tgaagcgcag 360
acgctcgcaa acgtgtattt ggccattgac aacaatttag aaattttacc agtcattaat 420
aaaatcgatt tgccaagcgc cgagccggag cgtgtccgcc aagaaatcga agacgtcatt 480
ggcctcgatg cctctgaagc ggtgctcgcc tccgcgaaag tcggcatcgg cgtcgaggac 540
attttagaac aaatcgtgga aaaaattcct gctccgtcag gcgatccgga cgcgccgttg 600
aaggcgctca tttttgattc actttatgac ccgtaccgcg gcgttgtcgc ctacgtccgt 660
atcgtcgatg gaacggttaa gccgggccag cgcattaaaa tgatgtcgac cggcaaagag 720
tttgaagtga ccgaagtcgg cgtgtttaca ccaaaaccaa aagttgtcga cgaactgatg 780
gtcggtgatg tcggctattt aactgcgtcg atcaaaaacg tacaagatac gcgcgtcggc 840
gatacgatta ccgatgccga acggccggct gctgagccac tccctggcta ccggaagctc 900
aatccgatgg tgttttgcgg catgtacccg atcgacacgg cgcgctacaa cgacttgcgc 960
gaagcgttag aaaagctgca gctcaacgat gcggcgcttc actttgaacc ggaaacgtcg 1020
caggcgctcg ggtttggctt tcgttgcggg tttctcggct tgcttcatat ggagattatc 1080
caagagcgga ttgaacgtga atttcatatc gatttaatta caacggcgcc gagcgttgtc 1140
tacaaagtat atttaacgga cggaacggaa gtcgatgtcg acaacccgac gaacatgccg 1200
gatccgcaaa aaatcgaccg catcgaagag ccgtatgtaa aagcgacgat tatggtgccg 1260
aacgactacg tcggaccggt gatggagctg tgccaaggaa agcgtggcac gttcgttgac 1320
atgcaatatt tagatgaaaa gcgggtcatg ttgatttacg atattccgct gtcggaaatc 1380
gtgtatgact ttttcgatgc gttaaagtcg aacacgaaag ggtatgcgtc gtttgactat 1440
gaattgatcg gttaccggcc gtccaatctt gtcaaaatgg atattttgtt gaatggcgaa 1500
aaaattgacg ctttatcgtt tattgttcac cgcgattcgg cttatgagcg cggcaaagtg 1560
atcgtcgaga agctgaaaga tttaattcca cgccaacagt ttgaagtgcc tgtgcaggcg 1620
gcgatcggca ataagatcat cgcccgttcg acgatcaagg cgctgcgtaa aaacgtgctc 1680
gccaaatgtt acggcggcga cgtgtcgcgg aaacggaaac tgcttgagaa acaaaaagaa 1740
ggaaagaaac ggatgaaaca aatcggttcg gtcgaagtgc cgcaggaagc gtttatggct 1800
gtcttgaaaa tcgacgacca gaaaaaa 1827
<210> 82
<211> 609
<212> PRT
<213> Geobacillus subterraneus
<400> 82
Met Asn Arg Glu Glu Arg Leu Lys Arg Gln Glu Arg Ile Arg Asn Phe
1 5 10 15
Ser Ile Ile Ala His Ile Asp His Gly Lys Ser Thr Leu Ala Asp Arg
20 25 30
Ile Leu Glu Lys Thr Gly Ala Leu Ser Glu Arg Glu Leu Arg Glu Gln
35 40 45
Thr Leu Asp Met Met Glu Leu Glu Arg Glu Arg Gly Ile Thr Ile Lys
50 55 60
Leu Asn Ala Val Gln Leu Thr Tyr Lys Ala Lys Asn Gly Glu Glu Tyr
65 70 75 80
Ile Phe His Leu Ile Asp Thr Pro Gly His Val Asp Phe Thr Tyr Glu
85 90 95
Val Ser Arg Ser Leu Ala Ala Cys Glu Gly Ala Ile Leu Val Val Asp
100 105 110
Ala Ala Gln Gly Ile Glu Ala Gln Thr Leu Ala Asn Val Tyr Leu Ala
115 120 125
Ile Asp Asn Asn Leu Glu Ile Leu Pro Val Ile Asn Lys Ile Asp Leu
130 135 140
Pro Ser Ala Glu Pro Glu Arg Val Arg Gln Glu Ile Glu Asp Val Ile
145 150 155 160
Gly Leu Asp Ala Ser Glu Ala Val Leu Ala Ser Ala Lys Val Gly Ile
165 170 175
Gly Val Glu Asp Ile Leu Glu Gln Ile Val Glu Lys Ile Pro Ala Pro
180 185 190
Ser Gly Asp Pro Asp Ala Pro Leu Lys Ala Leu Ile Phe Asp Ser Leu
195 200 205
Tyr Asp Pro Tyr Arg Gly Val Val Ala Tyr Val Arg Ile Val Asp Gly
210 215 220
Thr Val Lys Pro Gly Gln Arg Ile Lys Met Met Ser Thr Gly Lys Glu
225 230 235 240
Phe Glu Val Thr Glu Val Gly Val Phe Thr Pro Lys Pro Lys Val Val
245 250 255
Asp Glu Leu Met Val Gly Asp Val Gly Tyr Leu Thr Ala Ser Ile Lys
260 265 270
Asn Val Gln Asp Thr Arg Val Gly Asp Thr Ile Thr Asp Ala Glu Arg
275 280 285
Pro Ala Ala Glu Pro Leu Pro Gly Tyr Arg Lys Leu Asn Pro Met Val
290 295 300
Phe Cys Gly Met Tyr Pro Ile Asp Thr Ala Arg Tyr Asn Asp Leu Arg
305 310 315 320
Glu Ala Leu Glu Lys Leu Gln Leu Asn Asp Ala Ala Leu His Phe Glu
325 330 335
Pro Glu Thr Ser Gln Ala Leu Gly Phe Gly Phe Arg Cys Gly Phe Leu
340 345 350
Gly Leu Leu His Met Glu Ile Ile Gln Glu Arg Ile Glu Arg Glu Phe
355 360 365
His Ile Asp Leu Ile Thr Thr Ala Pro Ser Val Val Tyr Lys Val Tyr
370 375 380
Leu Thr Asp Gly Thr Glu Val Asp Val Asp Asn Pro Thr Asn Met Pro
385 390 395 400
Asp Pro Gln Lys Ile Asp Arg Ile Glu Glu Pro Tyr Val Lys Ala Thr
405 410 415
Ile Met Val Pro Asn Asp Tyr Val Gly Pro Val Met Glu Leu Cys Gln
420 425 430
Gly Lys Arg Gly Thr Phe Val Asp Met Gln Tyr Leu Asp Glu Lys Arg
435 440 445
Val Met Leu Ile Tyr Asp Ile Pro Leu Ser Glu Ile Val Tyr Asp Phe
450 455 460
Phe Asp Ala Leu Lys Ser Asn Thr Lys Gly Tyr Ala Ser Phe Asp Tyr
465 470 475 480
Glu Leu Ile Gly Tyr Arg Pro Ser Asn Leu Val Lys Met Asp Ile Leu
485 490 495
Leu Asn Gly Glu Lys Ile Asp Ala Leu Ser Phe Ile Val His Arg Asp
500 505 510
Ser Ala Tyr Glu Arg Gly Lys Val Ile Val Glu Lys Leu Lys Asp Leu
515 520 525
Ile Pro Arg Gln Gln Phe Glu Val Pro Val Gln Ala Ala Ile Gly Asn
530 535 540
Lys Ile Ile Ala Arg Ser Thr Ile Lys Ala Leu Arg Lys Asn Val Leu
545 550 555 560
Ala Lys Cys Tyr Gly Gly Asp Val Ser Arg Lys Arg Lys Leu Leu Glu
565 570 575
Lys Gln Lys Glu Gly Lys Lys Arg Met Lys Gln Ile Gly Ser Val Glu
580 585 590
Val Pro Gln Glu Ala Phe Met Ala Val Leu Lys Ile Asp Asp Gln Lys
595 600 605
Lys
<210> 83
<211> 555
<212> DNA
<213> Geobacillus subterraneus
<400> 83
atgatttcag tgaacgattt tcgcacaggg cttacgattg aggtcgacgg cgagatttgg 60
cgcgtccttg agttccagca tgttaagccg ggcaaagggg cggcgttcgt ccgttcgaag 120
ctgcgcaact tgcgtaccgg cgccattcaa gagcggacgt tccgcgctgg cgaaaaagta 180
aaccgggcac aaattgatac gcgcaaaatg caatatttat acgctaacgg cgacttgcat 240
gtctttatgg atatggaaac atacgaacaa atcgagctgc cagcgaaaca aattgagtat 300
gagctgaagt tcttaaaaga aaacatggaa gtatttatca tgatgtatca aggcgaaacg 360
atcggtgttg agctgccgaa caccgtcgag ttgaaagtcg ttgaaacaga gccgggcatc 420
aaaggtgaca cggcttccgg cggttcgaag ccggccaagc tcgaaaccgg tcttgtcgtt 480
caagtgccgt ttttcgtcaa tgaaggcgac acgctcatca ttaacacggc tgacggtacg 540
tacgtttcgc gggca 555
<210> 84
<211> 185
<212> PRT
<213> Geobacillus subterraneus
<400> 84
Met Ile Ser Val Asn Asp Phe Arg Thr Gly Leu Thr Ile Glu Val Asp
1 5 10 15
Gly Glu Ile Trp Arg Val Leu Glu Phe Gln His Val Lys Pro Gly Lys
20 25 30
Gly Ala Ala Phe Val Arg Ser Lys Leu Arg Asn Leu Arg Thr Gly Ala
35 40 45
Ile Gln Glu Arg Thr Phe Arg Ala Gly Glu Lys Val Asn Arg Ala Gln
50 55 60
Ile Asp Thr Arg Lys Met Gln Tyr Leu Tyr Ala Asn Gly Asp Leu His
65 70 75 80
Val Phe Met Asp Met Glu Thr Tyr Glu Gln Ile Glu Leu Pro Ala Lys
85 90 95
Gln Ile Glu Tyr Glu Leu Lys Phe Leu Lys Glu Asn Met Glu Val Phe
100 105 110
Ile Met Met Tyr Gln Gly Glu Thr Ile Gly Val Glu Leu Pro Asn Thr
115 120 125
Val Glu Leu Lys Val Val Glu Thr Glu Pro Gly Ile Lys Gly Asp Thr
130 135 140
Ala Ser Gly Gly Ser Lys Pro Ala Lys Leu Glu Thr Gly Leu Val Val
145 150 155 160
Gln Val Pro Phe Phe Val Asn Glu Gly Asp Thr Leu Ile Ile Asn Thr
165 170 175
Ala Asp Gly Thr Tyr Val Ser Arg Ala
180 185
<210> 85
<211> 1017
<212> DNA
<213> Geobacillus subterraneus
<400> 85
atggatccag ccgttatcaa cgacccgaaa aagttgcgcg attattcgaa agagcaggct 60
gatttgactg aaacggtgca aacgtaccgt gaatacaagt ccgttcgcag tcagctcgcg 120
gaagcgaagg ctatgctgga agaaaaactt gagccagagc tgcgcgagat ggtgaaagag 180
gaaattgatg agctcgaaga acgggaagaa gcgctcgttg agaagttgaa agtgttgctt 240
ttgccgaaag atccgaatga tgagaaaaac gtcattatgg aaattcgtgc cgccgccggt 300
ggcgaggaag ccgcgctgtt tgccggcgac ttgtaccgga tgtatacgcg ctatgcggag 360
tcgcaagggt ggaaaacgga agtgatcgaa gcaagcccaa caggtcttgg cggctataaa 420
gaaatcatct ttatggtcaa tgggaaaggg gcgtattcga agctgaagtt tgaaaacggc 480
gctcatcgcg tccaacgcgt cccggaaacg gaatcaggcg gacgcatcca tacatcgacg 540
gcaacggtcg cctgcttgcc ggaaatggaa gaagtcgaag tcgaaattca tgaaaaagac 600
attcgcgtcg atacgtacgc ctcgagcggg ccagggggac aaagcgtgaa cacgacgatg 660
tcagccgtac gcctcaccca tattccgacc ggcattgtcg ttacttgcca agacgaaaaa 720
tcgcaaatta aaaacaaaga aaaagcgatg aaagtgttgc gcgcccgcat ttacgacaaa 780
taccagcaag aagcgcgcgc cgagtatgac caaacgcgta agcaagcagt cggcaccggc 840
gatcgctcag agcgcatccg cacgtacaac ttcccgcaaa accgcgtcac tgaccaccgt 900
atcgggttga cgattcaaaa gcttgacctc gtgttagacg ggcagctcga tgaaattatc 960
gaggcgctca ttttagacga ccagtcgaaa aaactggagc aagcgaacga tgcgtcg 1017
<210> 86
<211> 339
<212> PRT
<213> Geobacillus subterraneus
<400> 86
Met Asp Pro Ala Val Ile Asn Asp Pro Lys Lys Leu Arg Asp Tyr Ser
1 5 10 15
Lys Glu Gln Ala Asp Leu Thr Glu Thr Val Gln Thr Tyr Arg Glu Tyr
20 25 30
Lys Ser Val Arg Ser Gln Leu Ala Glu Ala Lys Ala Met Leu Glu Glu
35 40 45
Lys Leu Glu Pro Glu Leu Arg Glu Met Val Lys Glu Glu Ile Asp Glu
50 55 60
Leu Glu Glu Arg Glu Glu Ala Leu Val Glu Lys Leu Lys Val Leu Leu
65 70 75 80
Leu Pro Lys Asp Pro Asn Asp Glu Lys Asn Val Ile Met Glu Ile Arg
85 90 95
Ala Ala Ala Gly Gly Glu Glu Ala Ala Leu Phe Ala Gly Asp Leu Tyr
100 105 110
Arg Met Tyr Thr Arg Tyr Ala Glu Ser Gln Gly Trp Lys Thr Glu Val
115 120 125
Ile Glu Ala Ser Pro Thr Gly Leu Gly Gly Tyr Lys Glu Ile Ile Phe
130 135 140
Met Val Asn Gly Lys Gly Ala Tyr Ser Lys Leu Lys Phe Glu Asn Gly
145 150 155 160
Ala His Arg Val Gln Arg Val Pro Glu Thr Glu Ser Gly Gly Arg Ile
165 170 175
His Thr Ser Thr Ala Thr Val Ala Cys Leu Pro Glu Met Glu Glu Val
180 185 190
Glu Val Glu Ile His Glu Lys Asp Ile Arg Val Asp Thr Tyr Ala Ser
195 200 205
Ser Gly Pro Gly Gly Gln Ser Val Asn Thr Thr Met Ser Ala Val Arg
210 215 220
Leu Thr His Ile Pro Thr Gly Ile Val Val Thr Cys Gln Asp Glu Lys
225 230 235 240
Ser Gln Ile Lys Asn Lys Glu Lys Ala Met Lys Val Leu Arg Ala Arg
245 250 255
Ile Tyr Asp Lys Tyr Gln Gln Glu Ala Arg Ala Glu Tyr Asp Gln Thr
260 265 270
Arg Lys Gln Ala Val Gly Thr Gly Asp Arg Ser Glu Arg Ile Arg Thr
275 280 285
Tyr Asn Phe Pro Gln Asn Arg Val Thr Asp His Arg Ile Gly Leu Thr
290 295 300
Ile Gln Lys Leu Asp Leu Val Leu Asp Gly Gln Leu Asp Glu Ile Ile
305 310 315 320
Glu Ala Leu Ile Leu Asp Asp Gln Ser Lys Lys Leu Glu Gln Ala Asn
325 330 335
Asp Ala Ser
<210> 87
<211> 981
<212> DNA
<213> Geobacillus subterraneus
<400> 87
atggccgcgc ccggcttttg ggatgaccag aaagcggcgc aggcgatcat ttccgaagcg 60
aatgcgctca aggaattagt cggcgagttt gaatcgctcg cggaacggtt cgacaacttg 120
gaagtgacgt atgagttgtt gaaagaggag ccggatgacg agctgcaggc tgaacttgtg 180
gaagaagcga aaaaattgac gaaagacttc agccagtttg agctgcagct gttgctcaac 240
gagccgtacg accaaaataa cgcgattttg gagcttcatc cgggtgcggg cggcacggaa 300
tcgcaagact gggcgtcgat gctgttgcgc atgtacacgc gctgggcgga gaaaaaagga 360
tttaaagtcg aaacactgga ttatctccca ggcgaggaag ccggggtgaa aagcgtcacc 420
ttgcttatca agggacataa tgcatacggc tacttaaagg cggaaaaagg ggtacaccgg 480
cttgtgcgca tctccccgtt tgacgcctca ggccgccgcc atacgtcgtt cgtgtcatgc 540
gaagtcgtgc cggagatgga cgataacatt gagattgaga tccgtccgga agagctgaaa 600
atcgacacgt accgctcaag cggtgcgggc gggcagcacg tcaacacgac cgactccgcg 660
gtgcgcatca cccacttgcc gaccggcatt gtcgttacgt gccaatcgga gcggtcgcaa 720
attaaaaacc gcgaaaaagc gatgaatatg ttaaaagcga agctgtatca aaagaaaatg 780
gaggaacagc aagctgaact cgccgagctg cgcggcgagc aaaaagaaat cggctggggc 840
agccaaatcc gctcctacgt cttccatccg tattcgcttg tcaaagacca tcggacgaat 900
gtggaggtcg gcaacgtgca agcggtgatg gatggggaaa tcgatgtgtt cattgacgcg 960
tatttgcgcg cgaaattgaa g 981
<210> 88
<211> 327
<212> PRT
<213> Geobacillus subterraneus
<400> 88
Met Ala Ala Pro Gly Phe Trp Asp Asp Gln Lys Ala Ala Gln Ala Ile
1 5 10 15
Ile Ser Glu Ala Asn Ala Leu Lys Glu Leu Val Gly Glu Phe Glu Ser
20 25 30
Leu Ala Glu Arg Phe Asp Asn Leu Glu Val Thr Tyr Glu Leu Leu Lys
35 40 45
Glu Glu Pro Asp Asp Glu Leu Gln Ala Glu Leu Val Glu Glu Ala Lys
50 55 60
Lys Leu Thr Lys Asp Phe Ser Gln Phe Glu Leu Gln Leu Leu Leu Asn
65 70 75 80
Glu Pro Tyr Asp Gln Asn Asn Ala Ile Leu Glu Leu His Pro Gly Ala
85 90 95
Gly Gly Thr Glu Ser Gln Asp Trp Ala Ser Met Leu Leu Arg Met Tyr
100 105 110
Thr Arg Trp Ala Glu Lys Lys Gly Phe Lys Val Glu Thr Leu Asp Tyr
115 120 125
Leu Pro Gly Glu Glu Ala Gly Val Lys Ser Val Thr Leu Leu Ile Lys
130 135 140
Gly His Asn Ala Tyr Gly Tyr Leu Lys Ala Glu Lys Gly Val His Arg
145 150 155 160
Leu Val Arg Ile Ser Pro Phe Asp Ala Ser Gly Arg Arg His Thr Ser
165 170 175
Phe Val Ser Cys Glu Val Val Pro Glu Met Asp Asp Asn Ile Glu Ile
180 185 190
Glu Ile Arg Pro Glu Glu Leu Lys Ile Asp Thr Tyr Arg Ser Ser Gly
195 200 205
Ala Gly Gly Gln His Val Asn Thr Thr Asp Ser Ala Val Arg Ile Thr
210 215 220
His Leu Pro Thr Gly Ile Val Val Thr Cys Gln Ser Glu Arg Ser Gln
225 230 235 240
Ile Lys Asn Arg Glu Lys Ala Met Asn Met Leu Lys Ala Lys Leu Tyr
245 250 255
Gln Lys Lys Met Glu Glu Gln Gln Ala Glu Leu Ala Glu Leu Arg Gly
260 265 270
Glu Gln Lys Glu Ile Gly Trp Gly Ser Gln Ile Arg Ser Tyr Val Phe
275 280 285
His Pro Tyr Ser Leu Val Lys Asp His Arg Thr Asn Val Glu Val Gly
290 295 300
Asn Val Gln Ala Val Met Asp Gly Glu Ile Asp Val Phe Ile Asp Ala
305 310 315 320
Tyr Leu Arg Ala Lys Leu Lys
325
<210> 89
<211> 555
<212> DNA
<213> Geobacillus subterraneus
<400> 89
atggcaaagc aagtgatcca acaggcgaaa gaaaaaatgg ataaagctgt gcaagcgttc 60
agccgcgagt tggcgaccgt ccgtgccggt cgggcgaacg cggggttgct tgagaaagta 120
accgttgact attacggtgt cgcaacgccg atcaaccagc tcgctacgat cagcgtgccg 180
gaagcgcgta tgcttgtcat tcagccgtat gacaaatcgg tcattaaaga aatggaaaaa 240
gcgattttag cgtcggactt aggagtgacg ccgtcgaatg acggatcggt tatccgcctt 300
gtcattccgc cgcttactga agaacgtcgc cgtgaactgg cgaagctcgt caaaaaatat 360
tcggaagaag cgaaagttgc ggtgcgcaac atccgtcgcg atgcaaacga tgagctgaaa 420
aaactcgaga aaaatagcga gattacggaa gatgagctgc gcagctatac cgacgaagtg 480
caaaagctga ccgacagcca tatcgccaaa attgacgcca tcacaaaaga gaaagaaaaa 540
gaagtgatgg aagta 555
<210> 90
<211> 185
<212> PRT
<213> Geobacillus subterraneus
<400> 90
Met Ala Lys Gln Val Ile Gln Gln Ala Lys Glu Lys Met Asp Lys Ala
1 5 10 15
Val Gln Ala Phe Ser Arg Glu Leu Ala Thr Val Arg Ala Gly Arg Ala
20 25 30
Asn Ala Gly Leu Leu Glu Lys Val Thr Val Asp Tyr Tyr Gly Val Ala
35 40 45
Thr Pro Ile Asn Gln Leu Ala Thr Ile Ser Val Pro Glu Ala Arg Met
50 55 60
Leu Val Ile Gln Pro Tyr Asp Lys Ser Val Ile Lys Glu Met Glu Lys
65 70 75 80
Ala Ile Leu Ala Ser Asp Leu Gly Val Thr Pro Ser Asn Asp Gly Ser
85 90 95
Val Ile Arg Leu Val Ile Pro Pro Leu Thr Glu Glu Arg Arg Arg Glu
100 105 110
Leu Ala Lys Leu Val Lys Lys Tyr Ser Glu Glu Ala Lys Val Ala Val
115 120 125
Arg Asn Ile Arg Arg Asp Ala Asn Asp Glu Leu Lys Lys Leu Glu Lys
130 135 140
Asn Ser Glu Ile Thr Glu Asp Glu Leu Arg Ser Tyr Thr Asp Glu Val
145 150 155 160
Gln Lys Leu Thr Asp Ser His Ile Ala Lys Ile Asp Ala Ile Thr Lys
165 170 175
Glu Lys Glu Lys Glu Val Met Glu Val
180 185
<210> 91
<211> 2676
<212> DNA
<213> Geobacillus subterraneus
<400> 91
atgagagttt ttttatataa aagaccaaag gggaggattg ttatgaaaaa gttaacatct 60
gccgaagtgc ggcgtatgtt tttgcagttt ttccaagaaa aaggccatgc ggtcgagccg 120
agcgcttcgc tcattcctgt cgatgacccg tcgttattat ggatcaacag cggtgtcgcg 180
acgctgaaaa aatattttga tggccgtatc atcccggaca acccgcgcat ttgcaatgcg 240
caaaaatcga tccgcacaaa cgacatcgaa aatgtcggga aaacggctcg ccaccatacg 300
ttttttgaaa tgctcggcaa cttttcgatc ggcgattatt tcaagcgtga agcgattcat 360
tgggcatggg agtttttaac aagtgaaaag tggattggtt ttgatccaga gcggttgtca 420
gtcactgttc atccggaaga cgaagaggcg tataacattt ggcgcaacga gatcggtctt 480
cctgaagagc ggattattcg tttagaagga aacttctggg atatcggtga aggcccgagc 540
ggtccgaaca cggaaatttt ttatgaccgc ggtgaagcgt tcggcaacga tccaaacgat 600
ccagaactgt atccaggcgg ggaaaatgac cgctacttag aagtatggaa tctcgtcttt 660
tcacagttca accataaccc ggacggcacg tacacgccgc tgccgaagaa aaacatcgat 720
accggcatgg gcttagagcg gatgtgctcg attttgcaag atgtaccgac gaactttgaa 780
actgatttgt tcatgccgat catccgcgcg actgagcaga tcgcgggtga gcaatacggc 840
aaagatccga ataaagacgt tgcttttaag gtcatcgctg accatattcg tgccgtgacg 900
tttgcggtcg gcgacggggc gctgccgtcg aacgaaggac gaggctatgt attgcgccgc 960
ctgcttcgcc gcgctgtgcg ctatgcgaaa caaatcggca ttgaccgtcc atttatgtat 1020
gagcttgttc cggttgtcgg tgaaattatg caagactatt atccggaagt gaaagaaaaa 1080
gccgatttca tcgcccgcgt cattcggacg gaagaagagc ggttccacga aacgcttcat 1140
gaagggctcg ccattttggc agaagtgatg gaaaaggcga aaaaacaagg aagcaccgtc 1200
attccaggag aagaggcgtt ccgcttgtac gatacgtacg gcttcccgct cgagctgacg 1260
gaagaatatg ctgctgaagc gggcatgtcg gtcgatcacg ccggttttga gcgcgagatg 1320
gagcgccagc gcgaacgggc ccgtgccgct cgccaagatg tcgattcgat gcaagtgcaa 1380
ggcggggtgc tcggcgacat taaagacgaa agccgttttg tcggctacga tgagctcgtc 1440
gtttcttcga cggtcattgc catcattaaa gacggacagc tcgtggagga agtcgggact 1500
ggcgaggaag cacaaatcat cgttgatgtg acgccgtttt acgccgaaag cggcggacaa 1560
atcgctgacc aaggtgtgtt tgaaggcgaa acgggaacag cggtcgtcaa agatgtgcaa 1620
aaagcaccga acggtcagca cctccattcg attgtcgtcg aacgcggtgc ggtgaaaaaa 1680
ggcgatcgct atacggcgcg cgtcgatgaa gtgaagcggt cgcaaatcgt gaaaaaccat 1740
acggcgaccc acttgcttca tcaagcgtta aaagacgttc ttggccgcca tgtcaaccag 1800
gccggatcac tcgttgcccc ggatcggctt cgctttgact ttactcattt cgggcaagtg 1860
aagcctgatg agctcgagcg cattgaggcg atcgtcaatg aacaaatttg gaagagtatt 1920
ccggtcgaca ttttttacaa accgctcgag gaagcaaaag cgatgggggc gatggcgctg 1980
tttggtgaaa aatacggcga tatcgtccgc gttgttaaag ttggcgacta cagcttagag 2040
ttgtgcggcg gctgccatgt gccgaataca gcggccattg ggttgtttaa aatcgtctcc 2100
gagtccggca tcggtgccgg cacgcgccgg attgaagcgg tgactgggga agcggcatac 2160
cgctttatga gcgaacagct tgctctgttg caagaagcgg cgcaaaagct gaaaacgagc 2220
ccgagagagc tgaatgcccg ccttgatggg ctgtttgccg aactgcgcca actgcagcgc 2280
gaaaatgagt cgcttgctgc ccgtctcgcc catatggagg cggaacacct cacccgtcaa 2340
gtgaaagagg tgggcggtgt gccggtatta gccgcaaaag tgcaggcgaa cgacatgaac 2400
caattgcggg cgatggctga tgacttgaag caaaaactag ggacggcggt catcgtgtta 2460
gcggccgtgc aaggtggcaa agtccaattg attgctgcgg tgactgatga cttagtgaaa 2520
aaaggatacc acgccggcaa actcgtcaaa gaagtggctt cacgttgcgg cggcggaggc 2580
ggcggacgtc ctgatatggc gcaggccggt gggaaggacg cgaacaaagt cggcgaagcg 2640
ctcgattatg tcgaaacatg ggtcaaatcc atttcc 2676
<210> 92
<211> 892
<212> PRT
<213> Geobacillus subterraneus
<400> 92
Met Arg Val Phe Leu Tyr Lys Arg Pro Lys Gly Arg Ile Val Met Lys
1 5 10 15
Lys Leu Thr Ser Ala Glu Val Arg Arg Met Phe Leu Gln Phe Phe Gln
20 25 30
Glu Lys Gly His Ala Val Glu Pro Ser Ala Ser Leu Ile Pro Val Asp
35 40 45
Asp Pro Ser Leu Leu Trp Ile Asn Ser Gly Val Ala Thr Leu Lys Lys
50 55 60
Tyr Phe Asp Gly Arg Ile Ile Pro Asp Asn Pro Arg Ile Cys Asn Ala
65 70 75 80
Gln Lys Ser Ile Arg Thr Asn Asp Ile Glu Asn Val Gly Lys Thr Ala
85 90 95
Arg His His Thr Phe Phe Glu Met Leu Gly Asn Phe Ser Ile Gly Asp
100 105 110
Tyr Phe Lys Arg Glu Ala Ile His Trp Ala Trp Glu Phe Leu Thr Ser
115 120 125
Glu Lys Trp Ile Gly Phe Asp Pro Glu Arg Leu Ser Val Thr Val His
130 135 140
Pro Glu Asp Glu Glu Ala Tyr Asn Ile Trp Arg Asn Glu Ile Gly Leu
145 150 155 160
Pro Glu Glu Arg Ile Ile Arg Leu Glu Gly Asn Phe Trp Asp Ile Gly
165 170 175
Glu Gly Pro Ser Gly Pro Asn Thr Glu Ile Phe Tyr Asp Arg Gly Glu
180 185 190
Ala Phe Gly Asn Asp Pro Asn Asp Pro Glu Leu Tyr Pro Gly Gly Glu
195 200 205
Asn Asp Arg Tyr Leu Glu Val Trp Asn Leu Val Phe Ser Gln Phe Asn
210 215 220
His Asn Pro Asp Gly Thr Tyr Thr Pro Leu Pro Lys Lys Asn Ile Asp
225 230 235 240
Thr Gly Met Gly Leu Glu Arg Met Cys Ser Ile Leu Gln Asp Val Pro
245 250 255
Thr Asn Phe Glu Thr Asp Leu Phe Met Pro Ile Ile Arg Ala Thr Glu
260 265 270
Gln Ile Ala Gly Glu Gln Tyr Gly Lys Asp Pro Asn Lys Asp Val Ala
275 280 285
Phe Lys Val Ile Ala Asp His Ile Arg Ala Val Thr Phe Ala Val Gly
290 295 300
Asp Gly Ala Leu Pro Ser Asn Glu Gly Arg Gly Tyr Val Leu Arg Arg
305 310 315 320
Leu Leu Arg Arg Ala Val Arg Tyr Ala Lys Gln Ile Gly Ile Asp Arg
325 330 335
Pro Phe Met Tyr Glu Leu Val Pro Val Val Gly Glu Ile Met Gln Asp
340 345 350
Tyr Tyr Pro Glu Val Lys Glu Lys Ala Asp Phe Ile Ala Arg Val Ile
355 360 365
Arg Thr Glu Glu Glu Arg Phe His Glu Thr Leu His Glu Gly Leu Ala
370 375 380
Ile Leu Ala Glu Val Met Glu Lys Ala Lys Lys Gln Gly Ser Thr Val
385 390 395 400
Ile Pro Gly Glu Glu Ala Phe Arg Leu Tyr Asp Thr Tyr Gly Phe Pro
405 410 415
Leu Glu Leu Thr Glu Glu Tyr Ala Ala Glu Ala Gly Met Ser Val Asp
420 425 430
His Ala Gly Phe Glu Arg Glu Met Glu Arg Gln Arg Glu Arg Ala Arg
435 440 445
Ala Ala Arg Gln Asp Val Asp Ser Met Gln Val Gln Gly Gly Val Leu
450 455 460
Gly Asp Ile Lys Asp Glu Ser Arg Phe Val Gly Tyr Asp Glu Leu Val
465 470 475 480
Val Ser Ser Thr Val Ile Ala Ile Ile Lys Asp Gly Gln Leu Val Glu
485 490 495
Glu Val Gly Thr Gly Glu Glu Ala Gln Ile Ile Val Asp Val Thr Pro
500 505 510
Phe Tyr Ala Glu Ser Gly Gly Gln Ile Ala Asp Gln Gly Val Phe Glu
515 520 525
Gly Glu Thr Gly Thr Ala Val Val Lys Asp Val Gln Lys Ala Pro Asn
530 535 540
Gly Gln His Leu His Ser Ile Val Val Glu Arg Gly Ala Val Lys Lys
545 550 555 560
Gly Asp Arg Tyr Thr Ala Arg Val Asp Glu Val Lys Arg Ser Gln Ile
565 570 575
Val Lys Asn His Thr Ala Thr His Leu Leu His Gln Ala Leu Lys Asp
580 585 590
Val Leu Gly Arg His Val Asn Gln Ala Gly Ser Leu Val Ala Pro Asp
595 600 605
Arg Leu Arg Phe Asp Phe Thr His Phe Gly Gln Val Lys Pro Asp Glu
610 615 620
Leu Glu Arg Ile Glu Ala Ile Val Asn Glu Gln Ile Trp Lys Ser Ile
625 630 635 640
Pro Val Asp Ile Phe Tyr Lys Pro Leu Glu Glu Ala Lys Ala Met Gly
645 650 655
Ala Met Ala Leu Phe Gly Glu Lys Tyr Gly Asp Ile Val Arg Val Val
660 665 670
Lys Val Gly Asp Tyr Ser Leu Glu Leu Cys Gly Gly Cys His Val Pro
675 680 685
Asn Thr Ala Ala Ile Gly Leu Phe Lys Ile Val Ser Glu Ser Gly Ile
690 695 700
Gly Ala Gly Thr Arg Arg Ile Glu Ala Val Thr Gly Glu Ala Ala Tyr
705 710 715 720
Arg Phe Met Ser Glu Gln Leu Ala Leu Leu Gln Glu Ala Ala Gln Lys
725 730 735
Leu Lys Thr Ser Pro Arg Glu Leu Asn Ala Arg Leu Asp Gly Leu Phe
740 745 750
Ala Glu Leu Arg Gln Leu Gln Arg Glu Asn Glu Ser Leu Ala Ala Arg
755 760 765
Leu Ala His Met Glu Ala Glu His Leu Thr Arg Gln Val Lys Glu Val
770 775 780
Gly Gly Val Pro Val Leu Ala Ala Lys Val Gln Ala Asn Asp Met Asn
785 790 795 800
Gln Leu Arg Ala Met Ala Asp Asp Leu Lys Gln Lys Leu Gly Thr Ala
805 810 815
Val Ile Val Leu Ala Ala Val Gln Gly Gly Lys Val Gln Leu Ile Ala
820 825 830
Ala Val Thr Asp Asp Leu Val Lys Lys Gly Tyr His Ala Gly Lys Leu
835 840 845
Val Lys Glu Val Ala Ser Arg Cys Gly Gly Gly Gly Gly Gly Arg Pro
850 855 860
Asp Met Ala Gln Ala Gly Gly Lys Asp Ala Asn Lys Val Gly Glu Ala
865 870 875 880
Leu Asp Tyr Val Glu Thr Trp Val Lys Ser Ile Ser
885 890
<210> 93
<211> 1671
<212> DNA
<213> Geobacillus subterraneus
<400> 93
atgaacattg tcggacaaat gaaagaacag ctgaaagagg aaattcgcca ggcggtggga 60
aaagccgggc tggtggcggc tgaggagctg ccagaagtat tgcttgaggt gccgcgcgaa 120
aaggctcatg gcgattattc gacgaatatc gccatgcagc tcgcccgcat cgcgaaaaag 180
ccaccgcggg caatcgccga agccatcgtt gaaaagtttg acgccgagcg tgtttcggtg 240
gcgcgcatcg aggtagccgg cccagggttt attaactttt acatggacaa tcgctatttg 300
acagcggttg tgccggcgat tttgcaagcg ggccaagcgt atggcgagtc gaatgtcggc 360
aaaggggaaa aagtgcaagt cgagttcgtc tcggctaacc cgaccggcaa cttgcattta 420
ggtcatgctc gcggtgcggc ggttggcgat tcacttagca atattttggc gaaagccgga 480
ttcgatgtga cgcgtgaata ttacattaat gatgccggca aacaaattta taacttggcg 540
aaatcagtcg aagcccgcta tttccaagcg ctcggtaccg atatgccgct gccggaggac 600
ggctattacg gtgacgacat cgtggaaatc ggcaaaaagc tcgccgatga atatggcgat 660
cggttcgtcc atgtggacga agaagaacga ctcgcctttt tccgcgaata cggcctccgt 720
tatgagctcg acaaaattaa aaacgatttg gctgccttcc gcgttccatt tgacgtttgg 780
tattcggaaa catcgcttta tgagagcggc aaaatcgatg aggcgctctc aacgctgcgt 840
gagcgcggtt acatttacga acaggacgga gccacatggt ttcgttcgac ggcgtttggc 900
gatgacaaag accgtgtgtt aatcaagcaa gacggaacgt atacgtattt gcttccggac 960
atcgcttacc atcaagataa gctgcggcgt gggttcacga agctaatcaa cgtctgggga 1020
gcggatcatc atggctacat cccgcgcatg aaagcggcga tcgctgcgct cggctacgat 1080
ccagaagcgc tcgaggtcga aattatccaa atggtgaact tataccaaaa cggcgagcgc 1140
gtcaaaatga gcaaacgtac tggcaaagcg gtgacgatgc gcgagctgat ggaagaagtc 1200
ggcgtcgatg ctgtccgcta cttcttcgct atgcgttcgg gcgatacgca tctcgatttt 1260
gatatggact tggctgttgc ccagtcgaat gaaaacccgg tctactatgt ccaatatgca 1320
catgcccgcg tctcaagcat tctccgtcaa gcaaaagagc atcaactgtc gtatgaaggc 1380
gacgtcgatc ttcatcatct cgtggaaaca gaaaaagaaa tcgagctgct caaagcgctt 1440
ggcgacttcc cggacgttgt cgctgaggcg gccttgaaac ggatgccaca tcgcgtcacc 1500
gcctatgcgt ttgatttggc gtcggcgctc cacagctttt acaatgcgga aaaagtgctt 1560
gacctagacc agatcgaaaa aacgaaagct cgtctcgcgc ttgtcaaggc ggtgcaaatc 1620
acgctgcaaa acgctctagc gttaatcggc gtctcagcgc cggaacaaat g 1671
<210> 94
<211> 557
<212> PRT
<213> Geobacillus subterraneus
<400> 94
Met Asn Ile Val Gly Gln Met Lys Glu Gln Leu Lys Glu Glu Ile Arg
1 5 10 15
Gln Ala Val Gly Lys Ala Gly Leu Val Ala Ala Glu Glu Leu Pro Glu
20 25 30
Val Leu Leu Glu Val Pro Arg Glu Lys Ala His Gly Asp Tyr Ser Thr
35 40 45
Asn Ile Ala Met Gln Leu Ala Arg Ile Ala Lys Lys Pro Pro Arg Ala
50 55 60
Ile Ala Glu Ala Ile Val Glu Lys Phe Asp Ala Glu Arg Val Ser Val
65 70 75 80
Ala Arg Ile Glu Val Ala Gly Pro Gly Phe Ile Asn Phe Tyr Met Asp
85 90 95
Asn Arg Tyr Leu Thr Ala Val Val Pro Ala Ile Leu Gln Ala Gly Gln
100 105 110
Ala Tyr Gly Glu Ser Asn Val Gly Lys Gly Glu Lys Val Gln Val Glu
115 120 125
Phe Val Ser Ala Asn Pro Thr Gly Asn Leu His Leu Gly His Ala Arg
130 135 140
Gly Ala Ala Val Gly Asp Ser Leu Ser Asn Ile Leu Ala Lys Ala Gly
145 150 155 160
Phe Asp Val Thr Arg Glu Tyr Tyr Ile Asn Asp Ala Gly Lys Gln Ile
165 170 175
Tyr Asn Leu Ala Lys Ser Val Glu Ala Arg Tyr Phe Gln Ala Leu Gly
180 185 190
Thr Asp Met Pro Leu Pro Glu Asp Gly Tyr Tyr Gly Asp Asp Ile Val
195 200 205
Glu Ile Gly Lys Lys Leu Ala Asp Glu Tyr Gly Asp Arg Phe Val His
210 215 220
Val Asp Glu Glu Glu Arg Leu Ala Phe Phe Arg Glu Tyr Gly Leu Arg
225 230 235 240
Tyr Glu Leu Asp Lys Ile Lys Asn Asp Leu Ala Ala Phe Arg Val Pro
245 250 255
Phe Asp Val Trp Tyr Ser Glu Thr Ser Leu Tyr Glu Ser Gly Lys Ile
260 265 270
Asp Glu Ala Leu Ser Thr Leu Arg Glu Arg Gly Tyr Ile Tyr Glu Gln
275 280 285
Asp Gly Ala Thr Trp Phe Arg Ser Thr Ala Phe Gly Asp Asp Lys Asp
290 295 300
Arg Val Leu Ile Lys Gln Asp Gly Thr Tyr Thr Tyr Leu Leu Pro Asp
305 310 315 320
Ile Ala Tyr His Gln Asp Lys Leu Arg Arg Gly Phe Thr Lys Leu Ile
325 330 335
Asn Val Trp Gly Ala Asp His His Gly Tyr Ile Pro Arg Met Lys Ala
340 345 350
Ala Ile Ala Ala Leu Gly Tyr Asp Pro Glu Ala Leu Glu Val Glu Ile
355 360 365
Ile Gln Met Val Asn Leu Tyr Gln Asn Gly Glu Arg Val Lys Met Ser
370 375 380
Lys Arg Thr Gly Lys Ala Val Thr Met Arg Glu Leu Met Glu Glu Val
385 390 395 400
Gly Val Asp Ala Val Arg Tyr Phe Phe Ala Met Arg Ser Gly Asp Thr
405 410 415
His Leu Asp Phe Asp Met Asp Leu Ala Val Ala Gln Ser Asn Glu Asn
420 425 430
Pro Val Tyr Tyr Val Gln Tyr Ala His Ala Arg Val Ser Ser Ile Leu
435 440 445
Arg Gln Ala Lys Glu His Gln Leu Ser Tyr Glu Gly Asp Val Asp Leu
450 455 460
His His Leu Val Glu Thr Glu Lys Glu Ile Glu Leu Leu Lys Ala Leu
465 470 475 480
Gly Asp Phe Pro Asp Val Val Ala Glu Ala Ala Leu Lys Arg Met Pro
485 490 495
His Arg Val Thr Ala Tyr Ala Phe Asp Leu Ala Ser Ala Leu His Ser
500 505 510
Phe Tyr Asn Ala Glu Lys Val Leu Asp Leu Asp Gln Ile Glu Lys Thr
515 520 525
Lys Ala Arg Leu Ala Leu Val Lys Ala Val Gln Ile Thr Leu Gln Asn
530 535 540
Ala Leu Ala Leu Ile Gly Val Ser Ala Pro Glu Gln Met
545 550 555
<210> 95
<211> 1323
<212> DNA
<213> Geobacillus subterraneus
<400> 95
atggacgtgt cgattattgg agggaatgtg tacgtgaaaa cgacgattgc tgaagtgaac 60
caatatgtag gtcaagaagt cacgatcggc gcttggttgg cgaacaagcg ctcgagcgga 120
aaaatcgcct ttttacagct gcgtgatggg actggcttta ttcaaggtgt agttgaaaaa 180
gcgaacgtct cagaagaggt atttcaacgt gcgaaaacgc tgacgcaaga aacgtcgctc 240
tatgtgaccg gcacggtgcg cgtcgacgag cgttcaccgt tcggttatga gctttcggtg 300
acgaacatac aggtcatcaa tgaagcggtc gattatccga ttacgccaaa agaacacggt 360
gtcgagtttt taatggatca tcgtcacctt tggcttcgtt cgcggcgcca acatgcgatc 420
atgaaaatcc gcaacgaatt gatccgtgcg acgtatgagt tttttaacga acgtggcttc 480
gtcaaagtcg atgcgccgat tttgactggc agcgcaccgg aaggaacgac cgagctgttc 540
catacgaagt attttgacga ggatgcctat ttatcgcaaa gcggccagct atatatggaa 600
gcagcagcca tggcgctcgg taaagtgttt tcgttcggtc cgacattccg tgccgaaaag 660
tcgaaaacgc gccgccattt gatcgaattt tggatgatcg agcctgaaat ggcgttttac 720
gaatttgaag acaatttgcg gctgcaagaa gagtatgtct cttatctcgt acagtcggtg 780
cttagccgtt gccaacttga gctcgggcgc cttggacgcg acgtcaccaa gcttgagctt 840
gtcaagccgc cgtttccgcg tctaacgtat gacgaagcga tcaagctgct gcatgacaaa 900
gggtttaccg atatcgaatg gggcgatgac ttcggtgcgc cgcatgagac agccatcgct 960
gaaagcttcg acaagccggt gtttatcact cactacccga cgtcgttaaa gccgttttat 1020
atgcagccag atccgaaccg tccggacgtc gtgctatgtg ctgatttaat cgcgccggag 1080
ggatacgggg agattatcgg cggttccgag cgcattcatg attatgagct gctcaagcag 1140
cgtctcgagg agcatcattt gccgcttgaa gcatatgaat ggtatttaga tttgcgcaaa 1200
tacggttccg tgccgcactc cggattcggg ctcggcctcg agcgaacggt tgcttggatt 1260
tgcggcgttg agcatgtacg cgagacgatc ccgtttccgc ggttgctcaa ccgtctatac 1320
ccg 1323
<210> 96
<211> 441
<212> PRT
<213> Geobacillus subterraneus
<400> 96
Met Asp Val Ser Ile Ile Gly Gly Asn Val Tyr Val Lys Thr Thr Ile
1 5 10 15
Ala Glu Val Asn Gln Tyr Val Gly Gln Glu Val Thr Ile Gly Ala Trp
20 25 30
Leu Ala Asn Lys Arg Ser Ser Gly Lys Ile Ala Phe Leu Gln Leu Arg
35 40 45
Asp Gly Thr Gly Phe Ile Gln Gly Val Val Glu Lys Ala Asn Val Ser
50 55 60
Glu Glu Val Phe Gln Arg Ala Lys Thr Leu Thr Gln Glu Thr Ser Leu
65 70 75 80
Tyr Val Thr Gly Thr Val Arg Val Asp Glu Arg Ser Pro Phe Gly Tyr
85 90 95
Glu Leu Ser Val Thr Asn Ile Gln Val Ile Asn Glu Ala Val Asp Tyr
100 105 110
Pro Ile Thr Pro Lys Glu His Gly Val Glu Phe Leu Met Asp His Arg
115 120 125
His Leu Trp Leu Arg Ser Arg Arg Gln His Ala Ile Met Lys Ile Arg
130 135 140
Asn Glu Leu Ile Arg Ala Thr Tyr Glu Phe Phe Asn Glu Arg Gly Phe
145 150 155 160
Val Lys Val Asp Ala Pro Ile Leu Thr Gly Ser Ala Pro Glu Gly Thr
165 170 175
Thr Glu Leu Phe His Thr Lys Tyr Phe Asp Glu Asp Ala Tyr Leu Ser
180 185 190
Gln Ser Gly Gln Leu Tyr Met Glu Ala Ala Ala Met Ala Leu Gly Lys
195 200 205
Val Phe Ser Phe Gly Pro Thr Phe Arg Ala Glu Lys Ser Lys Thr Arg
210 215 220
Arg His Leu Ile Glu Phe Trp Met Ile Glu Pro Glu Met Ala Phe Tyr
225 230 235 240
Glu Phe Glu Asp Asn Leu Arg Leu Gln Glu Glu Tyr Val Ser Tyr Leu
245 250 255
Val Gln Ser Val Leu Ser Arg Cys Gln Leu Glu Leu Gly Arg Leu Gly
260 265 270
Arg Asp Val Thr Lys Leu Glu Leu Val Lys Pro Pro Phe Pro Arg Leu
275 280 285
Thr Tyr Asp Glu Ala Ile Lys Leu Leu His Asp Lys Gly Phe Thr Asp
290 295 300
Ile Glu Trp Gly Asp Asp Phe Gly Ala Pro His Glu Thr Ala Ile Ala
305 310 315 320
Glu Ser Phe Asp Lys Pro Val Phe Ile Thr His Tyr Pro Thr Ser Leu
325 330 335
Lys Pro Phe Tyr Met Gln Pro Asp Pro Asn Arg Pro Asp Val Val Leu
340 345 350
Cys Ala Asp Leu Ile Ala Pro Glu Gly Tyr Gly Glu Ile Ile Gly Gly
355 360 365
Ser Glu Arg Ile His Asp Tyr Glu Leu Leu Lys Gln Arg Leu Glu Glu
370 375 380
His His Leu Pro Leu Glu Ala Tyr Glu Trp Tyr Leu Asp Leu Arg Lys
385 390 395 400
Tyr Gly Ser Val Pro His Ser Gly Phe Gly Leu Gly Leu Glu Arg Thr
405 410 415
Val Ala Trp Ile Cys Gly Val Glu His Val Arg Glu Thr Ile Pro Phe
420 425 430
Pro Arg Leu Leu Asn Arg Leu Tyr Pro
435 440
<210> 97
<211> 1359
<212> DNA
<213> Geobacillus subterraneus
<400> 97
atgtttcaaa cacttgagct tcgtcataaa gtggcgaagg cggtgcgcaa ctttttagac 60
ggcgaacgct ttttagaagt ggagacgcca atgttgacga aaagcacacc ggaaggggcg 120
cgcgattatt tagtgccaag ccgcgttcat ccgggggaat tttacgcctt gccgcagtcg 180
ccgcaaattt ttaagcagct tttgatggtc ggcggttttg aacgctatta ccaaatcact 240
cgttgcttcc gcgatgaaga tttgcgcgct gaccgccagc cagagtttac gcaaattgac 300
attgaaatgt cgtttgtcga ccaagaagac atcatcgatt taaccgaacg gatgatggcg 360
gcggtcgtca aagcaactaa agggattgac attccgcgcc catttccacg catcacgtat 420
gacgaagcga tgagccgtta cggttccgat aagccggacg tacgttttgg ccttgagctt 480
gtcgatgtgt cggaagcggt ccgcggctcc gcgtttcaag tgttcgcccg cgccgttgag 540
caaggtggtc aagtgaaggc aatcaacgta aaaggagcgg cgagccgtta ttcgcgtaaa 600
gacattgacg cgttagcgga gtttgccggc cgctacggag cgaaagggct cgcttggtta 660
aaagttgaag gcggggagct gaaagggccg atcgccaagt ttttcgtcga tgatgagcaa 720
acagcgctgc gccagctgct tgctgccgaa gatggggatt tgctgttgtt tgttgctgac 780
gagaaggcga ttgtcgcggc ggctcttggt gcgttgcggt taaagctcgg caaagagctt 840
ggcttgatcg atgaaacgaa gctcgctttt ttatgggtaa cagattggcc gcttttagag 900
tacgacgaag aagaaggccg ctattacgcc gcccaccatc cgtttacgat gccggtgcgt 960
gacgatatcc cgctgcttga gacaaaccca ggcgctgttc gggcgcaggc gtatgattta 1020
gtgttaaacg gctatgagct tggcggcggt tcgctccgta tttttgagcg cgatgtacaa 1080
gaaaaaatgt tccgcgctct aggatttgac caggaagagg cgcgccgcca gtttggcttc 1140
ctgcttgagg cgtttgaata tggcactccg ccgcatggcg gtatcgccct cggcctcgat 1200
cgacttgtga tgctcttagc tgggcgcaca aacttgcgcg atacgatcgc cttcccgaaa 1260
actgcgagcg ccagctgcct gcttactgaa gcgccgggac cggtcagtga aaaacaactg 1320
aaagagttgc atttggctgt ggtgcttccc gaccagcaa 1359
<210> 98
<211> 453
<212> PRT
<213> Geobacillus subterraneus
<400> 98
Met Phe Gln Thr Leu Glu Leu Arg His Lys Val Ala Lys Ala Val Arg
1 5 10 15
Asn Phe Leu Asp Gly Glu Arg Phe Leu Glu Val Glu Thr Pro Met Leu
20 25 30
Thr Lys Ser Thr Pro Glu Gly Ala Arg Asp Tyr Leu Val Pro Ser Arg
35 40 45
Val His Pro Gly Glu Phe Tyr Ala Leu Pro Gln Ser Pro Gln Ile Phe
50 55 60
Lys Gln Leu Leu Met Val Gly Gly Phe Glu Arg Tyr Tyr Gln Ile Thr
65 70 75 80
Arg Cys Phe Arg Asp Glu Asp Leu Arg Ala Asp Arg Gln Pro Glu Phe
85 90 95
Thr Gln Ile Asp Ile Glu Met Ser Phe Val Asp Gln Glu Asp Ile Ile
100 105 110
Asp Leu Thr Glu Arg Met Met Ala Ala Val Val Lys Ala Thr Lys Gly
115 120 125
Ile Asp Ile Pro Arg Pro Phe Pro Arg Ile Thr Tyr Asp Glu Ala Met
130 135 140
Ser Arg Tyr Gly Ser Asp Lys Pro Asp Val Arg Phe Gly Leu Glu Leu
145 150 155 160
Val Asp Val Ser Glu Ala Val Arg Gly Ser Ala Phe Gln Val Phe Ala
165 170 175
Arg Ala Val Glu Gln Gly Gly Gln Val Lys Ala Ile Asn Val Lys Gly
180 185 190
Ala Ala Ser Arg Tyr Ser Arg Lys Asp Ile Asp Ala Leu Ala Glu Phe
195 200 205
Ala Gly Arg Tyr Gly Ala Lys Gly Leu Ala Trp Leu Lys Val Glu Gly
210 215 220
Gly Glu Leu Lys Gly Pro Ile Ala Lys Phe Phe Val Asp Asp Glu Gln
225 230 235 240
Thr Ala Leu Arg Gln Leu Leu Ala Ala Glu Asp Gly Asp Leu Leu Leu
245 250 255
Phe Val Ala Asp Glu Lys Ala Ile Val Ala Ala Ala Leu Gly Ala Leu
260 265 270
Arg Leu Lys Leu Gly Lys Glu Leu Gly Leu Ile Asp Glu Thr Lys Leu
275 280 285
Ala Phe Leu Trp Val Thr Asp Trp Pro Leu Leu Glu Tyr Asp Glu Glu
290 295 300
Glu Gly Arg Tyr Tyr Ala Ala His His Pro Phe Thr Met Pro Val Arg
305 310 315 320
Asp Asp Ile Pro Leu Leu Glu Thr Asn Pro Gly Ala Val Arg Ala Gln
325 330 335
Ala Tyr Asp Leu Val Leu Asn Gly Tyr Glu Leu Gly Gly Gly Ser Leu
340 345 350
Arg Ile Phe Glu Arg Asp Val Gln Glu Lys Met Phe Arg Ala Leu Gly
355 360 365
Phe Asp Gln Glu Glu Ala Arg Arg Gln Phe Gly Phe Leu Leu Glu Ala
370 375 380
Phe Glu Tyr Gly Thr Pro Pro His Gly Gly Ile Ala Leu Gly Leu Asp
385 390 395 400
Arg Leu Val Met Leu Leu Ala Gly Arg Thr Asn Leu Arg Asp Thr Ile
405 410 415
Ala Phe Pro Lys Thr Ala Ser Ala Ser Cys Leu Leu Thr Glu Ala Pro
420 425 430
Gly Pro Val Ser Glu Lys Gln Leu Lys Glu Leu His Leu Ala Val Val
435 440 445
Leu Pro Asp Gln Gln
450
<210> 99
<211> 1419
<212> DNA
<213> Geobacillus subterraneus
<400> 99
atgaaaggaa gagcgaatat gagcagtatc cgactttata atacgttgac gcgaaaaaag 60
gaaacgtttg agccgctcga accgaacaaa gtgaaaatgt atgtatgtgg cccgacggtc 120
tataattata ttcatatcgg caatgctcgc gccgctatcg tctttgatac gatccgccgt 180
tatttagagt tccgcggtta tgatgtgacg tatgtatcca actttactga tgtcgacgac 240
aagctaatca gggcggcccg cgagcttggt gagagcgtgc cggcgatcgc cgagcggttt 300
attgaggcgt attttgagga cattgaggcg ctcggctgca aaaaagcaga tatccatccg 360
cgcgtgacgg aaaatatcga aacgattatc gaattcattc aagcgctcat tgacaaaggc 420
tatgcgtacg aagtcgatgg tgacgtatac tatcggacgc gcaagtttga tggctacggc 480
aaattgtcgc atcagtcgat cgatgagcta caagcggggg cgcgcatcga agttggggaa 540
aagaaagatg atccactcga ttttgctctt tggaaagcag cgaaagaagg agagatttct 600
tgggacagcc catgggggaa agggcggccc ggctggcata tcgaatgttc agcgatggcg 660
cgcaaatatt taggagatac gatcgacatt catgctggcg gccaagactt aacgtttcca 720
caccatgaaa acgaaattgc ccaatcggaa gcactgaccg gcaaaccgtt tgcgaaatat 780
tggctgcaca atgggtattt aaatattaac aatgaaaaaa tgtccaagtc gcttggcaac 840
tttgtacttg ttcacgatat catccggcag attgacccac aagtgttgcg tttctttatg 900
ctgtcggtgc actatcgcca cccgatcaac tatagcgagg agctgcttga gagcgctcgg 960
cgtggtctcg aacgcttgag gacagcatac ggtaatttgc agcaccggct tggggcgagc 1020
acgaacttaa ccgataacga cggcgagtgg ctttcgcgcc tcgcggatat ccgcgcctcg 1080
ttcattcgtg aaatggacga tgatttcaac acagcaaacg gcattgcggt cttgttcgag 1140
ctcgccaaac aagcgaactt gtatttgcag gagaaaacga catccgagaa tgtcattcac 1200
gcgtttttgc gcgaatttga gcagctgatg gatgtactcg gccttacttt gaaacaagag 1260
gagttgcttg acgaagaaat tgaggcgctg atccgccagc gcaatgaagc gcggaaaaat 1320
cgtgactttg ccttagccga ccgcatccgc gacgagttga aagcaaaaaa tatcattttg 1380
gaagatacgc cgcaagggac gagatggaaa cggggatcg 1419
<210> 100
<211> 473
<212> PRT
<213> Geobacillus subterraneus
<400> 100
Met Lys Gly Arg Ala Asn Met Ser Ser Ile Arg Leu Tyr Asn Thr Leu
1 5 10 15
Thr Arg Lys Lys Glu Thr Phe Glu Pro Leu Glu Pro Asn Lys Val Lys
20 25 30
Met Tyr Val Cys Gly Pro Thr Val Tyr Asn Tyr Ile His Ile Gly Asn
35 40 45
Ala Arg Ala Ala Ile Val Phe Asp Thr Ile Arg Arg Tyr Leu Glu Phe
50 55 60
Arg Gly Tyr Asp Val Thr Tyr Val Ser Asn Phe Thr Asp Val Asp Asp
65 70 75 80
Lys Leu Ile Arg Ala Ala Arg Glu Leu Gly Glu Ser Val Pro Ala Ile
85 90 95
Ala Glu Arg Phe Ile Glu Ala Tyr Phe Glu Asp Ile Glu Ala Leu Gly
100 105 110
Cys Lys Lys Ala Asp Ile His Pro Arg Val Thr Glu Asn Ile Glu Thr
115 120 125
Ile Ile Glu Phe Ile Gln Ala Leu Ile Asp Lys Gly Tyr Ala Tyr Glu
130 135 140
Val Asp Gly Asp Val Tyr Tyr Arg Thr Arg Lys Phe Asp Gly Tyr Gly
145 150 155 160
Lys Leu Ser His Gln Ser Ile Asp Glu Leu Gln Ala Gly Ala Arg Ile
165 170 175
Glu Val Gly Glu Lys Lys Asp Asp Pro Leu Asp Phe Ala Leu Trp Lys
180 185 190
Ala Ala Lys Glu Gly Glu Ile Ser Trp Asp Ser Pro Trp Gly Lys Gly
195 200 205
Arg Pro Gly Trp His Ile Glu Cys Ser Ala Met Ala Arg Lys Tyr Leu
210 215 220
Gly Asp Thr Ile Asp Ile His Ala Gly Gly Gln Asp Leu Thr Phe Pro
225 230 235 240
His His Glu Asn Glu Ile Ala Gln Ser Glu Ala Leu Thr Gly Lys Pro
245 250 255
Phe Ala Lys Tyr Trp Leu His Asn Gly Tyr Leu Asn Ile Asn Asn Glu
260 265 270
Lys Met Ser Lys Ser Leu Gly Asn Phe Val Leu Val His Asp Ile Ile
275 280 285
Arg Gln Ile Asp Pro Gln Val Leu Arg Phe Phe Met Leu Ser Val His
290 295 300
Tyr Arg His Pro Ile Asn Tyr Ser Glu Glu Leu Leu Glu Ser Ala Arg
305 310 315 320
Arg Gly Leu Glu Arg Leu Arg Thr Ala Tyr Gly Asn Leu Gln His Arg
325 330 335
Leu Gly Ala Ser Thr Asn Leu Thr Asp Asn Asp Gly Glu Trp Leu Ser
340 345 350
Arg Leu Ala Asp Ile Arg Ala Ser Phe Ile Arg Glu Met Asp Asp Asp
355 360 365
Phe Asn Thr Ala Asn Gly Ile Ala Val Leu Phe Glu Leu Ala Lys Gln
370 375 380
Ala Asn Leu Tyr Leu Gln Glu Lys Thr Thr Ser Glu Asn Val Ile His
385 390 395 400
Ala Phe Leu Arg Glu Phe Glu Gln Leu Met Asp Val Leu Gly Leu Thr
405 410 415
Leu Lys Gln Glu Glu Leu Leu Asp Glu Glu Ile Glu Ala Leu Ile Arg
420 425 430
Gln Arg Asn Glu Ala Arg Lys Asn Arg Asp Phe Ala Leu Ala Asp Arg
435 440 445
Ile Arg Asp Glu Leu Lys Ala Lys Asn Ile Ile Leu Glu Asp Thr Pro
450 455 460
Gln Gly Thr Arg Trp Lys Arg Gly Ser
465 470
<210> 101
<211> 1488
<212> DNA
<213> Geobacillus subterraneus
<400> 101
atggaattgg aggtttggac gatggcaaaa aacgtgcgcg tgcgctatgc gccgagcccg 60
actggccatt tgcatatcgg tggggcacgg acagcgctgt ttaactattt gtttgcccgc 120
cattacggcg gaaaaatgat cgtccgcatc gaagatacgg atattgaacg gaacgttgaa 180
ggcggcgaag agtcgcagct tgaaaactta aaatggcttg gcatcgatta tgacgaatcg 240
attgataagg acggcggata tgggccgtat cgtcagacgg aacggctcga tatctatcgg 300
aagtatgtga acgagctgct tgaacaaggg catgcgtata aatgtttttg tacaccggaa 360
gagctcgagc gggaacgtga ggagcaacgg gcggcaggta ttgctgctcc gcaatacagc 420
ggcaaatgcc gccatttaac gccggagcaa gttgccgagc ttgaagcaca aggaaaaccg 480
tatacgatcc gcttgaaagt gccggaaggg aaaacgtatg aagtagatga tttagtgcgc 540
ggtaaagtga cgtttgaatc gaaagacatc ggcgattggg tcattgtgaa ggcgaacggt 600
attccgacgt acaactttgc cgttgtcatt gatgaccatt tgatggaaat cagccatgtg 660
ttccgcggtg aggagcattt atccaacacg ccgaaacagc taatggtgta cgaatatttc 720
ggttgggagc caccgcaatt cgcccatatg acattgattg tcaacgagca gcggaaaaag 780
ctatccaagc gcgatgaatc gattatccag ttcgtgtcgc aatataaaga gctcggctat 840
ttgccggagg cgatgttcaa ctttttcgcc cttcttggct ggtcgccgga aggagaagaa 900
gaaattttta cgaaggacga gctcatccgc atttttgatg tcgcccggct gtcgaaatcg 960
ccgtcgatgt ttgatacgaa aaagctgaca tggatgaaca accaatatat caaaaagctg 1020
gatctcgaca ggcttgtcga gctggcgttg ccgcatttag tgaaagccgg acgcctgccg 1080
gcagatatga gtgatgagca gcggcaatgg gcacgcgatt tgattgcctt gtaccaagag 1140
caaatgagct acggtgcgga gatcgtttcg ctgtccgagc tgttctttaa agaagaagtc 1200
gaatacgaag acgaagcccg ccaagtgctc gccgaagaac aagtaccgga tgtgctctcc 1260
gcctttttgg cgaatgtgcg tgagcttgag ccgtttacgg cggatgagat taaagcagcg 1320
atcaaagcag tgcaaaaatc gacagggcaa aaaggcaaga agctgtttat gccgattcgc 1380
gccgcagtga ctgggcaaac acacggaccg gaactgccgt ttgccatcca actgcttggc 1440
aaacaaaagg tgattgaacg gctcgaacgg gcactgcatg aaaaattt 1488
<210> 102
<211> 496
<212> PRT
<213> Geobacillus subterraneus
<400> 102
Met Glu Leu Glu Val Trp Thr Met Ala Lys Asn Val Arg Val Arg Tyr
1 5 10 15
Ala Pro Ser Pro Thr Gly His Leu His Ile Gly Gly Ala Arg Thr Ala
20 25 30
Leu Phe Asn Tyr Leu Phe Ala Arg His Tyr Gly Gly Lys Met Ile Val
35 40 45
Arg Ile Glu Asp Thr Asp Ile Glu Arg Asn Val Glu Gly Gly Glu Glu
50 55 60
Ser Gln Leu Glu Asn Leu Lys Trp Leu Gly Ile Asp Tyr Asp Glu Ser
65 70 75 80
Ile Asp Lys Asp Gly Gly Tyr Gly Pro Tyr Arg Gln Thr Glu Arg Leu
85 90 95
Asp Ile Tyr Arg Lys Tyr Val Asn Glu Leu Leu Glu Gln Gly His Ala
100 105 110
Tyr Lys Cys Phe Cys Thr Pro Glu Glu Leu Glu Arg Glu Arg Glu Glu
115 120 125
Gln Arg Ala Ala Gly Ile Ala Ala Pro Gln Tyr Ser Gly Lys Cys Arg
130 135 140
His Leu Thr Pro Glu Gln Val Ala Glu Leu Glu Ala Gln Gly Lys Pro
145 150 155 160
Tyr Thr Ile Arg Leu Lys Val Pro Glu Gly Lys Thr Tyr Glu Val Asp
165 170 175
Asp Leu Val Arg Gly Lys Val Thr Phe Glu Ser Lys Asp Ile Gly Asp
180 185 190
Trp Val Ile Val Lys Ala Asn Gly Ile Pro Thr Tyr Asn Phe Ala Val
195 200 205
Val Ile Asp Asp His Leu Met Glu Ile Ser His Val Phe Arg Gly Glu
210 215 220
Glu His Leu Ser Asn Thr Pro Lys Gln Leu Met Val Tyr Glu Tyr Phe
225 230 235 240
Gly Trp Glu Pro Pro Gln Phe Ala His Met Thr Leu Ile Val Asn Glu
245 250 255
Gln Arg Lys Lys Leu Ser Lys Arg Asp Glu Ser Ile Ile Gln Phe Val
260 265 270
Ser Gln Tyr Lys Glu Leu Gly Tyr Leu Pro Glu Ala Met Phe Asn Phe
275 280 285
Phe Ala Leu Leu Gly Trp Ser Pro Glu Gly Glu Glu Glu Ile Phe Thr
290 295 300
Lys Asp Glu Leu Ile Arg Ile Phe Asp Val Ala Arg Leu Ser Lys Ser
305 310 315 320
Pro Ser Met Phe Asp Thr Lys Lys Leu Thr Trp Met Asn Asn Gln Tyr
325 330 335
Ile Lys Lys Leu Asp Leu Asp Arg Leu Val Glu Leu Ala Leu Pro His
340 345 350
Leu Val Lys Ala Gly Arg Leu Pro Ala Asp Met Ser Asp Glu Gln Arg
355 360 365
Gln Trp Ala Arg Asp Leu Ile Ala Leu Tyr Gln Glu Gln Met Ser Tyr
370 375 380
Gly Ala Glu Ile Val Ser Leu Ser Glu Leu Phe Phe Lys Glu Glu Val
385 390 395 400
Glu Tyr Glu Asp Glu Ala Arg Gln Val Leu Ala Glu Glu Gln Val Pro
405 410 415
Asp Val Leu Ser Ala Phe Leu Ala Asn Val Arg Glu Leu Glu Pro Phe
420 425 430
Thr Ala Asp Glu Ile Lys Ala Ala Ile Lys Ala Val Gln Lys Ser Thr
435 440 445
Gly Gln Lys Gly Lys Lys Leu Phe Met Pro Ile Arg Ala Ala Val Thr
450 455 460
Gly Gln Thr His Gly Pro Glu Leu Pro Phe Ala Ile Gln Leu Leu Gly
465 470 475 480
Lys Gln Lys Val Ile Glu Arg Leu Glu Arg Ala Leu His Glu Lys Phe
485 490 495
<210> 103
<211> 1398
<212> DNA
<213> Geobacillus subterraneus
<400> 103
atggaggagg atgatgacat ggctgcaaca atggaagaaa tcgttgccca cgccaagcat 60
cgcggcttcg tgtttccggg gtcggaaatt tacggtgggc tggcgaacac atgggattac 120
ggtccgctcg gtgtcgagct gaaaaataac attaaacggg cgtggtggaa aaagttcgtc 180
caagaatcgc cacacaatgt cggtttggac gctgccattt taatgaaccc aaaaacgtgg 240
gaagcatccg gccatttagg caacttcaac gatccgatgg tcgactgcaa acagtgtaaa 300
gcgcgtcatc gcgccgacaa gctgattgag caggcacttg aagaaaaagg aattgagatg 360
gtcgttgacg gtttgccgct tgccaagatg gaagagctta tccgtgaata cgacatcgct 420
tgtccagaat gcggcagtcg tgactttacg aacgtgcgtc agtttaattt aatgttcaaa 480
acataccaag gtgtcaccga atcaagcgct aacgaaattt atttgcgccc ggagacggcc 540
caaggtattt ttgtcaactt taaaaacgtc cagcgcacga tgcgcaaaaa attaccgttt 600
ggcatcgcgc aaatcggaaa aagtttccgc aacgaaatta cgccagggaa ctttacgttc 660
cgcacacgtg aatttgaaca aatggagctt gagtttttct gcaaaccggg cgaagagctg 720
aaatggttcg actactggaa acaattttgc aaggaatggc tgttgtcgct cggcatgaac 780
gaagaacata tccgcctgcg cgaccatacg aaagaagaat tatcccacta tagtaatgcg 840
acgactgata tcgagtatca gttcccgttc ggctggggcg agctctgggg tattgcgtcg 900
cgcaccgatt acgacttaaa acagcatatg gaacactccg gtgaggattt ccattatctt 960
gaccaagaaa cgaatgagcg ctacatcccg tactgcattg agccgtcgct cggtgccgac 1020
cgtgtcacgc tcgcgtttat gattgacgcc tatgacgagg aagagctcga agacggcacg 1080
acccggacag ttatgcattt gcatccagcg cttgcgccgt acaaagcagc tgtcttgccg 1140
ttatcgaaaa agctgggtga cggagcgcgc cgaatttatg aagagctcgc gaagcatttc 1200
atggtcgact acgatgaaac aggttcgatt ggcaagcggt atcgtcgtca agatgaaatc 1260
ggcacgccgt tttgtatcac gtacgacttt gagtccgagc aagatggcca agtaaccgtt 1320
cgtgaccgtg acacgatgga acaagtgcgg ttgccgattg gggagctcaa agcctttttg 1380
gataaaaaaa ttgccttt 1398
<210> 104
<211> 466
<212> PRT
<213> Geobacillus subterraneus
<400> 104
Met Glu Glu Asp Asp Asp Met Ala Ala Thr Met Glu Glu Ile Val Ala
1 5 10 15
His Ala Lys His Arg Gly Phe Val Phe Pro Gly Ser Glu Ile Tyr Gly
20 25 30
Gly Leu Ala Asn Thr Trp Asp Tyr Gly Pro Leu Gly Val Glu Leu Lys
35 40 45
Asn Asn Ile Lys Arg Ala Trp Trp Lys Lys Phe Val Gln Glu Ser Pro
50 55 60
His Asn Val Gly Leu Asp Ala Ala Ile Leu Met Asn Pro Lys Thr Trp
65 70 75 80
Glu Ala Ser Gly His Leu Gly Asn Phe Asn Asp Pro Met Val Asp Cys
85 90 95
Lys Gln Cys Lys Ala Arg His Arg Ala Asp Lys Leu Ile Glu Gln Ala
100 105 110
Leu Glu Glu Lys Gly Ile Glu Met Val Val Asp Gly Leu Pro Leu Ala
115 120 125
Lys Met Glu Glu Leu Ile Arg Glu Tyr Asp Ile Ala Cys Pro Glu Cys
130 135 140
Gly Ser Arg Asp Phe Thr Asn Val Arg Gln Phe Asn Leu Met Phe Lys
145 150 155 160
Thr Tyr Gln Gly Val Thr Glu Ser Ser Ala Asn Glu Ile Tyr Leu Arg
165 170 175
Pro Glu Thr Ala Gln Gly Ile Phe Val Asn Phe Lys Asn Val Gln Arg
180 185 190
Thr Met Arg Lys Lys Leu Pro Phe Gly Ile Ala Gln Ile Gly Lys Ser
195 200 205
Phe Arg Asn Glu Ile Thr Pro Gly Asn Phe Thr Phe Arg Thr Arg Glu
210 215 220
Phe Glu Gln Met Glu Leu Glu Phe Phe Cys Lys Pro Gly Glu Glu Leu
225 230 235 240
Lys Trp Phe Asp Tyr Trp Lys Gln Phe Cys Lys Glu Trp Leu Leu Ser
245 250 255
Leu Gly Met Asn Glu Glu His Ile Arg Leu Arg Asp His Thr Lys Glu
260 265 270
Glu Leu Ser His Tyr Ser Asn Ala Thr Thr Asp Ile Glu Tyr Gln Phe
275 280 285
Pro Phe Gly Trp Gly Glu Leu Trp Gly Ile Ala Ser Arg Thr Asp Tyr
290 295 300
Asp Leu Lys Gln His Met Glu His Ser Gly Glu Asp Phe His Tyr Leu
305 310 315 320
Asp Gln Glu Thr Asn Glu Arg Tyr Ile Pro Tyr Cys Ile Glu Pro Ser
325 330 335
Leu Gly Ala Asp Arg Val Thr Leu Ala Phe Met Ile Asp Ala Tyr Asp
340 345 350
Glu Glu Glu Leu Glu Asp Gly Thr Thr Arg Thr Val Met His Leu His
355 360 365
Pro Ala Leu Ala Pro Tyr Lys Ala Ala Val Leu Pro Leu Ser Lys Lys
370 375 380
Leu Gly Asp Gly Ala Arg Arg Ile Tyr Glu Glu Leu Ala Lys His Phe
385 390 395 400
Met Val Asp Tyr Asp Glu Thr Gly Ser Ile Gly Lys Arg Tyr Arg Arg
405 410 415
Gln Asp Glu Ile Gly Thr Pro Phe Cys Ile Thr Tyr Asp Phe Glu Ser
420 425 430
Glu Gln Asp Gly Gln Val Thr Val Arg Asp Arg Asp Thr Met Glu Gln
435 440 445
Val Arg Leu Pro Ile Gly Glu Leu Lys Ala Phe Leu Asp Lys Lys Ile
450 455 460
Ala Phe
465
<210> 105
<211> 1278
<212> DNA
<213> Geobacillus subterraneus
<400> 105
atggcttttc aaattccaag agggacacaa gatttattac cgggtgaaac ggaaaaatgg 60
caatatgtcg aacaagtggc ccgcgacctg tgtagacggt acggctatga agaaatacgg 120
acgccgattt ttgaacatac ggagctgttt ttacgtggcg ttggtgatac gaccgatatc 180
gtccaaaaag agatgtacac gtttgaagac aaagggggcc gtgcgttgac gctccgtccg 240
gaaggaaccg caccggtcgt gcgggcgttc gtcgagcata agctgtacgg cagcccgaat 300
cagccggtca agttgtatta tgcgggacca atgttccgtt atgagcggcc ggaagccgga 360
cggttccgcc aattcgtcca gtttggtgtt gaggcaattg gcagcagtga tccggcgatt 420
gacgccgagg tgatggcgtt agcgatgcat atttataagg cgcttggttt aaaacacatc 480
cggctcgtaa tcaacagttt aggcgatgta gacagccgcc gggcgcatcg cgaagcgctt 540
gtccgccatt tttctgaccg cattcatgaa ctgtgcccgg actgtcaggc gcggcttgag 600
acgaatccgc tccgcattct cgattgtaaa aaggaccgcg atcatgaact gatggcgtca 660
gcaccgtcga ttttagacta tttgaatgac gaatcgcgcg cgtattttga gaaggtgaag 720
caatatttaa cgatgcttga catcccgttt gtcattgact cgcggctcgt gcgcggcctc 780
gattattaca accatacgac gtttgaaatt atgagcgagg ctgaaggatt cggcgcagcg 840
gcgactcttt gcggcggcgg acgctataac gggcttgtgc aagaaattgg cggcccggaa 900
acgcctggca tcggctttgc gttaagcatt gaacggctgc tggcggcgct tgaagcggaa 960
gggattgaac tgccgatcca tcgaggaatc gattgctatg ttgtcgctgt cggtgagcgg 1020
gcaaaagatg aaactgtccg cctcgtttac gaattgcgcc gtgccggcct gcgtgtggag 1080
caagactatt taggtcgaaa aatgaaggca cagctgaagg cagctgaccg tcttggcgca 1140
tcattcgttg ccatcatcgg cgacgaggag ctggaaaaac agacagcagc tgtgaaacac 1200
atggcgagcg gcgagcaaac tgatgtgccg cttggagagt tggcgtcctt tttaatagaa 1260
cgaacaaaac gggaggag 1278
<210> 106
<211> 426
<212> PRT
<213> Geobacillus subterraneus
<400> 106
Met Ala Phe Gln Ile Pro Arg Gly Thr Gln Asp Leu Leu Pro Gly Glu
1 5 10 15
Thr Glu Lys Trp Gln Tyr Val Glu Gln Val Ala Arg Asp Leu Cys Arg
20 25 30
Arg Tyr Gly Tyr Glu Glu Ile Arg Thr Pro Ile Phe Glu His Thr Glu
35 40 45
Leu Phe Leu Arg Gly Val Gly Asp Thr Thr Asp Ile Val Gln Lys Glu
50 55 60
Met Tyr Thr Phe Glu Asp Lys Gly Gly Arg Ala Leu Thr Leu Arg Pro
65 70 75 80
Glu Gly Thr Ala Pro Val Val Arg Ala Phe Val Glu His Lys Leu Tyr
85 90 95
Gly Ser Pro Asn Gln Pro Val Lys Leu Tyr Tyr Ala Gly Pro Met Phe
100 105 110
Arg Tyr Glu Arg Pro Glu Ala Gly Arg Phe Arg Gln Phe Val Gln Phe
115 120 125
Gly Val Glu Ala Ile Gly Ser Ser Asp Pro Ala Ile Asp Ala Glu Val
130 135 140
Met Ala Leu Ala Met His Ile Tyr Lys Ala Leu Gly Leu Lys His Ile
145 150 155 160
Arg Leu Val Ile Asn Ser Leu Gly Asp Val Asp Ser Arg Arg Ala His
165 170 175
Arg Glu Ala Leu Val Arg His Phe Ser Asp Arg Ile His Glu Leu Cys
180 185 190
Pro Asp Cys Gln Ala Arg Leu Glu Thr Asn Pro Leu Arg Ile Leu Asp
195 200 205
Cys Lys Lys Asp Arg Asp His Glu Leu Met Ala Ser Ala Pro Ser Ile
210 215 220
Leu Asp Tyr Leu Asn Asp Glu Ser Arg Ala Tyr Phe Glu Lys Val Lys
225 230 235 240
Gln Tyr Leu Thr Met Leu Asp Ile Pro Phe Val Ile Asp Ser Arg Leu
245 250 255
Val Arg Gly Leu Asp Tyr Tyr Asn His Thr Thr Phe Glu Ile Met Ser
260 265 270
Glu Ala Glu Gly Phe Gly Ala Ala Ala Thr Leu Cys Gly Gly Gly Arg
275 280 285
Tyr Asn Gly Leu Val Gln Glu Ile Gly Gly Pro Glu Thr Pro Gly Ile
290 295 300
Gly Phe Ala Leu Ser Ile Glu Arg Leu Leu Ala Ala Leu Glu Ala Glu
305 310 315 320
Gly Ile Glu Leu Pro Ile His Arg Gly Ile Asp Cys Tyr Val Val Ala
325 330 335
Val Gly Glu Arg Ala Lys Asp Glu Thr Val Arg Leu Val Tyr Glu Leu
340 345 350
Arg Arg Ala Gly Leu Arg Val Glu Gln Asp Tyr Leu Gly Arg Lys Met
355 360 365
Lys Ala Gln Leu Lys Ala Ala Asp Arg Leu Gly Ala Ser Phe Val Ala
370 375 380
Ile Ile Gly Asp Glu Glu Leu Glu Lys Gln Thr Ala Ala Val Lys His
385 390 395 400
Met Ala Ser Gly Glu Gln Thr Asp Val Pro Leu Gly Glu Leu Ala Ser
405 410 415
Phe Leu Ile Glu Arg Thr Lys Arg Glu Glu
420 425
<210> 107
<211> 2772
<212> DNA
<213> Geobacillus subterraneus
<400> 107
atggactaca aagagacgct gctcatgccg caaacggagt tcccgatgcg tggcaacttg 60
ccgaagcggg agccggaaat gcaaaaaaaa tgggaggaaa tggacattta ccggaaagtg 120
caggagcgga cgaaaggacg gccgctgttt gtgctgcacg acggcccgcc atacgccaac 180
ggtgatattc atatgggcca tgcattaaat aaaattttaa aagatattat cgtccgctac 240
aagtcgatga gcggcttttg tgcgccgtat gtgcctggct gggatacaca tggcttaccg 300
attgaaacgg cactgacgaa gcaaggtgtc gaccgcaaat cgatgagtgt cgctgagttc 360
cgcaagctgt gcgaacaata cgcgtatgag caaatcgaca accagcgcca acagtttaaa 420
cggctcgggg tgcggggcga ttgggacaac ccgtacatta cgctcaagcc ggaatacgaa 480
gcccagcaaa ttaaagtgtt cggtgaaatg gcgaaaaaag ggctcattta taaagggctg 540
aagccggtgt attggtcgcc gtcgagcgaa tcggcgctcg ccgaagcgga aatcgaatat 600
aaagacaaac ggtcgccgtc gatttatgtc gcgttcccag ttaaagatgg taaaggtgtg 660
cttcaagggg atgaacgaat cgtcatttgg acgacgacac cgtggacgat tccagcgaac 720
ttggcgatcg ccgttcaccc ggatttggac tactatattg tcgaagcaaa cgggcaaaaa 780
tacgttgttg ctgcggcctt ggcggaatcg gtagcgaaag aagtcggctg ggaggcatgg 840
tccgtcgtca aaacggtaaa aggaaaagaa cttgagtacg tagtcgccaa acatccgttt 900
tacgagcgcg actcgcttgt cgtctgcggc gagcacgtca cgaccgacgc cggtaccggc 960
tgcgttcata cggcaccagg acacggggaa gacgacttta tcgtcggaca aaaatacggg 1020
cttccggttc tttgcccggt tgatgagcgc ggctatatga cagaagaagc gcctggattt 1080
gcagggatgt tttacgacga ggcgaacaaa gcgattacac aaaagctcga ggaagttgga 1140
gcgctcctta agctcagctt cattacccac tcgtatccgc atgattggcg gacgaagcaa 1200
ccgacaattt tccgagcgac gacacaatgg tttgcctcca ttgataaaat tcgtgatcaa 1260
cttcttgatg ccatcaagga aacgaaatgg gtgccagaat ggggagaaat ccgcatccat 1320
aacatggtgc gcgaccgcgg tgactggtgc atctcccgcc aacgcgcttg gggcgtgcca 1380
attccggtct tttacggcga aaacggcgag ccgatcatca cagatgagac gatcgagcac 1440
gtgtcaaacc tattccgcca gtacggctcg aatgtttggt ttgagcgtga ggcgaaagac 1500
ttattgccgg aaggattcac ccatccgtcc agcccgaacg gcctctttac gaaagagacg 1560
gatattatgg acgtctggtt tgactccggt tcgtcgcatc aagccgtgct tgttgaacgc 1620
gatgacctag agcgtccggc tgatttatac ttagaaggat ctgaccaata tcgcggctgg 1680
tttaactcgt cgctgtctac agccgttgcc gtcaccggaa aagcaccgta taaaggggtg 1740
ttaagccatg gcttcgtttt agacggcgaa gggcgaaaaa tgagcaaatc gctcggcaac 1800
gtcgtcgtgc cggccaaagt catggaacag ctcggtgccg acattttacg cctttgggtc 1860
gcctcggttg actatcaggc ggatgtacgc atttccgata acattttaaa acaagtgtcc 1920
gaagtgtatc ggaaaatccg caatacgttc cgctttatgc tcggcaactt gtttgatttt 1980
gacccgaatc aaaacgctgt gccggttggg gagcttggcg aagtcgatcg ctacatgtta 2040
gcgaaattaa ataaactcat cgctaaagtg aaaaaggcgt atgacagcta tgattttgct 2100
gctgtttatc atgagatgaa ccatttctgc accgtcgagt taagcgcatt ttatttggat 2160
atggcgaaag acattttgta catcgaagcg gccgattgtc gtgcccgccg tgcggtgcag 2220
acggtgctgt atgaaacggt tgtcgccttg gcgaagctca ttgcgccgat tttgccgcac 2280
acggccgatg aagtgtggga gcatatcccg aaccggaaag agcaagtgga aagcgtccag 2340
ctcaccgaca tgccggagtc aatggccatc gatggtgaag aagcgctgct tgcgaaatgg 2400
gatgcgttta tggatgtacg agatgacatt ttaaaagcgc tcgagaatgc gcgtaatgaa 2460
aaagtgatcg gtaagtcgct cacggcgagc gtcactgttt acccgaaaga cgaagtgcgg 2520
gcgcttttgg cttcgatcaa cgaggacttg cgccaacttc tcatcgtttc cgcgttttcg 2580
gtcgccgatg aatcgtatga cgccgcgcca gccgaagcag aacggctcaa ccatgtggcc 2640
gtcatcgttc gcccggcgga aggtgagacg tgcgaacgtt gctggacggt gacaccggac 2700
gtcggacgcg atgagtccca cccgacgctt tgtccgcgct gcgcacatat tgtgaacgaa 2760
cattattcgg ca 2772
<210> 108
<211> 924
<212> PRT
<213> Geobacillus subterraneus
<400> 108
Met Asp Tyr Lys Glu Thr Leu Leu Met Pro Gln Thr Glu Phe Pro Met
1 5 10 15
Arg Gly Asn Leu Pro Lys Arg Glu Pro Glu Met Gln Lys Lys Trp Glu
20 25 30
Glu Met Asp Ile Tyr Arg Lys Val Gln Glu Arg Thr Lys Gly Arg Pro
35 40 45
Leu Phe Val Leu His Asp Gly Pro Pro Tyr Ala Asn Gly Asp Ile His
50 55 60
Met Gly His Ala Leu Asn Lys Ile Leu Lys Asp Ile Ile Val Arg Tyr
65 70 75 80
Lys Ser Met Ser Gly Phe Cys Ala Pro Tyr Val Pro Gly Trp Asp Thr
85 90 95
His Gly Leu Pro Ile Glu Thr Ala Leu Thr Lys Gln Gly Val Asp Arg
100 105 110
Lys Ser Met Ser Val Ala Glu Phe Arg Lys Leu Cys Glu Gln Tyr Ala
115 120 125
Tyr Glu Gln Ile Asp Asn Gln Arg Gln Gln Phe Lys Arg Leu Gly Val
130 135 140
Arg Gly Asp Trp Asp Asn Pro Tyr Ile Thr Leu Lys Pro Glu Tyr Glu
145 150 155 160
Ala Gln Gln Ile Lys Val Phe Gly Glu Met Ala Lys Lys Gly Leu Ile
165 170 175
Tyr Lys Gly Leu Lys Pro Val Tyr Trp Ser Pro Ser Ser Glu Ser Ala
180 185 190
Leu Ala Glu Ala Glu Ile Glu Tyr Lys Asp Lys Arg Ser Pro Ser Ile
195 200 205
Tyr Val Ala Phe Pro Val Lys Asp Gly Lys Gly Val Leu Gln Gly Asp
210 215 220
Glu Arg Ile Val Ile Trp Thr Thr Thr Pro Trp Thr Ile Pro Ala Asn
225 230 235 240
Leu Ala Ile Ala Val His Pro Asp Leu Asp Tyr Tyr Ile Val Glu Ala
245 250 255
Asn Gly Gln Lys Tyr Val Val Ala Ala Ala Leu Ala Glu Ser Val Ala
260 265 270
Lys Glu Val Gly Trp Glu Ala Trp Ser Val Val Lys Thr Val Lys Gly
275 280 285
Lys Glu Leu Glu Tyr Val Val Ala Lys His Pro Phe Tyr Glu Arg Asp
290 295 300
Ser Leu Val Val Cys Gly Glu His Val Thr Thr Asp Ala Gly Thr Gly
305 310 315 320
Cys Val His Thr Ala Pro Gly His Gly Glu Asp Asp Phe Ile Val Gly
325 330 335
Gln Lys Tyr Gly Leu Pro Val Leu Cys Pro Val Asp Glu Arg Gly Tyr
340 345 350
Met Thr Glu Glu Ala Pro Gly Phe Ala Gly Met Phe Tyr Asp Glu Ala
355 360 365
Asn Lys Ala Ile Thr Gln Lys Leu Glu Glu Val Gly Ala Leu Leu Lys
370 375 380
Leu Ser Phe Ile Thr His Ser Tyr Pro His Asp Trp Arg Thr Lys Gln
385 390 395 400
Pro Thr Ile Phe Arg Ala Thr Thr Gln Trp Phe Ala Ser Ile Asp Lys
405 410 415
Ile Arg Asp Gln Leu Leu Asp Ala Ile Lys Glu Thr Lys Trp Val Pro
420 425 430
Glu Trp Gly Glu Ile Arg Ile His Asn Met Val Arg Asp Arg Gly Asp
435 440 445
Trp Cys Ile Ser Arg Gln Arg Ala Trp Gly Val Pro Ile Pro Val Phe
450 455 460
Tyr Gly Glu Asn Gly Glu Pro Ile Ile Thr Asp Glu Thr Ile Glu His
465 470 475 480
Val Ser Asn Leu Phe Arg Gln Tyr Gly Ser Asn Val Trp Phe Glu Arg
485 490 495
Glu Ala Lys Asp Leu Leu Pro Glu Gly Phe Thr His Pro Ser Ser Pro
500 505 510
Asn Gly Leu Phe Thr Lys Glu Thr Asp Ile Met Asp Val Trp Phe Asp
515 520 525
Ser Gly Ser Ser His Gln Ala Val Leu Val Glu Arg Asp Asp Leu Glu
530 535 540
Arg Pro Ala Asp Leu Tyr Leu Glu Gly Ser Asp Gln Tyr Arg Gly Trp
545 550 555 560
Phe Asn Ser Ser Leu Ser Thr Ala Val Ala Val Thr Gly Lys Ala Pro
565 570 575
Tyr Lys Gly Val Leu Ser His Gly Phe Val Leu Asp Gly Glu Gly Arg
580 585 590
Lys Met Ser Lys Ser Leu Gly Asn Val Val Val Pro Ala Lys Val Met
595 600 605
Glu Gln Leu Gly Ala Asp Ile Leu Arg Leu Trp Val Ala Ser Val Asp
610 615 620
Tyr Gln Ala Asp Val Arg Ile Ser Asp Asn Ile Leu Lys Gln Val Ser
625 630 635 640
Glu Val Tyr Arg Lys Ile Arg Asn Thr Phe Arg Phe Met Leu Gly Asn
645 650 655
Leu Phe Asp Phe Asp Pro Asn Gln Asn Ala Val Pro Val Gly Glu Leu
660 665 670
Gly Glu Val Asp Arg Tyr Met Leu Ala Lys Leu Asn Lys Leu Ile Ala
675 680 685
Lys Val Lys Lys Ala Tyr Asp Ser Tyr Asp Phe Ala Ala Val Tyr His
690 695 700
Glu Met Asn His Phe Cys Thr Val Glu Leu Ser Ala Phe Tyr Leu Asp
705 710 715 720
Met Ala Lys Asp Ile Leu Tyr Ile Glu Ala Ala Asp Cys Arg Ala Arg
725 730 735
Arg Ala Val Gln Thr Val Leu Tyr Glu Thr Val Val Ala Leu Ala Lys
740 745 750
Leu Ile Ala Pro Ile Leu Pro His Thr Ala Asp Glu Val Trp Glu His
755 760 765
Ile Pro Asn Arg Lys Glu Gln Val Glu Ser Val Gln Leu Thr Asp Met
770 775 780
Pro Glu Ser Met Ala Ile Asp Gly Glu Glu Ala Leu Leu Ala Lys Trp
785 790 795 800
Asp Ala Phe Met Asp Val Arg Asp Asp Ile Leu Lys Ala Leu Glu Asn
805 810 815
Ala Arg Asn Glu Lys Val Ile Gly Lys Ser Leu Thr Ala Ser Val Thr
820 825 830
Val Tyr Pro Lys Asp Glu Val Arg Ala Leu Leu Ala Ser Ile Asn Glu
835 840 845
Asp Leu Arg Gln Leu Leu Ile Val Ser Ala Phe Ser Val Ala Asp Glu
850 855 860
Ser Tyr Asp Ala Ala Pro Ala Glu Ala Glu Arg Leu Asn His Val Ala
865 870 875 880
Val Ile Val Arg Pro Ala Glu Gly Glu Thr Cys Glu Arg Cys Trp Thr
885 890 895
Val Thr Pro Asp Val Gly Arg Asp Glu Ser His Pro Thr Leu Cys Pro
900 905 910
Arg Cys Ala His Ile Val Asn Glu His Tyr Ser Ala
915 920
<210> 109
<211> 2433
<212> DNA
<213> Geobacillus subterraneus
<400> 109
atgaggagga gtgcgacgat gagtttcaac catcgcgaaa ttgagaaaaa gtggcaggat 60
tattgggaac agcataaaac gttccgcacc ccggatgaaa gcgataaacc gaagttttac 120
gtgttggata tgtttccgta tccgtctggc gctggcttgc acgtcggcca tccggaaggg 180
tatacggcga ctgatatttt ggcgcgcatg aagcggatgc aagggtacaa tgtccttcac 240
ccgatggggt gggacgcgtt cggattgccg gcagaacaat atgcgctcga taccggcaac 300
gacccggccg aatttacgca aaaaaacatc gacaacttcc gccggcaaat taagtcgctt 360
ggtttttcgt atgactggga tcgggaaatt aacacgactg atccgaacta ttacaaatgg 420
acgcaatgga ttttcttgaa gctgtatgaa aaagggctcg cctacatgga cgaagtaccg 480
gtcaactggt gtccggcgct tggcaccgtg ctggcgaacg aagaagtcat caacggccgg 540
agcgagcgcg gtgggcatcc ggtcatccgc aagccaatgc ggcaatggat gctgaaaatt 600
accgcctatg ccgaccggct gctcgaagat ttggaggagc ttgactggcc ggaaagcatt 660
aaagaaatgc aacgcaactg gatcggccgt tcggaaggag cggaaattga gtttgctgtc 720
gacggccatg acgagtcgtt cacggtattt acgacgcggc cagatacgct gtttggcgcc 780
acgtacgcag tgttggctcc ggaacatccg cttgttgaga aaattacaac gccggagcaa 840
aaaccagccg ttgatgctta cttaaaagaa gtgcaaagca aaagcgacct cgagcgcacc 900
gacttggcga aagaaaaaac aggcgtgttc actggtgcgt acgccatcca tccagttacc 960
ggcgacaagc tgccgatttg gatcgccgat tacgtgttga tgggctacgg cactggggcg 1020
atcatggctg taccggcgca tgatgagcgc gactacgagt ttgcgaaaac attcaacttg 1080
ccgatcaaag aagtcgttgc cggcgggaat gtcgaaaacg agccgtacac tggcgacggg 1140
gagcacatca actctgagtt tttgaacggc ttgaacaaac aagaagcgat cgaaaaaatg 1200
atcgcctggc ttgaagaaaa cggaaaagga caaaagaaag tgtcgtaccg gctgcgcgac 1260
tggttgttta gccgccaacg ctactggggt gagccgattc cggtcatcca ttgggaagat 1320
gggacgatga cgacggtgcc ggaagaagaa ttgccgcttg tcttgccgaa aacggatgaa 1380
attaaaccgt cgggaacggg tgaatcgccg ctcgccaaca tcgaagaatg ggtcaatgtt 1440
gtcgatccga aaaccgggaa aaaagggcgg cgtgaaacaa acacgatgcc gcaatgggcg 1500
ggaagctgct ggtattattt gcgctacatc gacccgcata acgacaaaca gctcgccgat 1560
ccggaaaagt tgaaacaatg gctgccggtt gacgtctaca tcggcggggc ggagcatgcg 1620
gtcttgcact tgctgtacgc tcgcttctgg cataaagtgt tgtacgacct tggcatcgtg 1680
ccgacgaaag agccgttcca aaagctgttt aaccaaggga tgatcttagg cgaaaacaat 1740
gaaaaaatga gcaaatcgaa aggcaatgtc gtcaacccgg atgatatcgt cgagagccat 1800
ggcgcggata cgttgcggct gtatgaaatg tttatggggc cgcttgaagc gtcgatcgcc 1860
tggtcgacga aagggcttga cggagcgcgc cgtttcttag agcgcgtctg gcgtctgttt 1920
gtcaccgaag atggtcaact gaacccgaac atcgttgacg agccagcgaa cgatacgctc 1980
gagcgcgtct accatcaaac ggtgaaaaaa gtgacggaag actacgaagc gctgcgcttc 2040
aacaccgcca tttcgcagct gatggtgttc attaacgaag cgtataaagc ggagcagatg 2100
aaaaaagaat atatggaagg gttcgtcaag ctcttatcgc cggtttgccc gcatattggc 2160
gaagagctct ggcaaaagct cggccatact gacaccatcg cctatgaacc atggccgaca 2220
tatgacgaag cgaaactcgt cgaagatgtc gttgaaatcg tgatccaaat caacggcaaa 2280
gtgcgggcga aactgaacgt gccggcggac ttatcgaaag aggcgctaga agaacgggcg 2340
ctcgccgatg aaaaaattaa agagcagctt gcagggaaaa cggtgcgtaa ggtgatcact 2400
gtccctggta agctcgtcaa tatcgtcgcc aac 2433
<210> 110
<211> 811
<212> PRT
<213> Geobacillus subterraneus
<400> 110
Met Arg Arg Ser Ala Thr Met Ser Phe Asn His Arg Glu Ile Glu Lys
1 5 10 15
Lys Trp Gln Asp Tyr Trp Glu Gln His Lys Thr Phe Arg Thr Pro Asp
20 25 30
Glu Ser Asp Lys Pro Lys Phe Tyr Val Leu Asp Met Phe Pro Tyr Pro
35 40 45
Ser Gly Ala Gly Leu His Val Gly His Pro Glu Gly Tyr Thr Ala Thr
50 55 60
Asp Ile Leu Ala Arg Met Lys Arg Met Gln Gly Tyr Asn Val Leu His
65 70 75 80
Pro Met Gly Trp Asp Ala Phe Gly Leu Pro Ala Glu Gln Tyr Ala Leu
85 90 95
Asp Thr Gly Asn Asp Pro Ala Glu Phe Thr Gln Lys Asn Ile Asp Asn
100 105 110
Phe Arg Arg Gln Ile Lys Ser Leu Gly Phe Ser Tyr Asp Trp Asp Arg
115 120 125
Glu Ile Asn Thr Thr Asp Pro Asn Tyr Tyr Lys Trp Thr Gln Trp Ile
130 135 140
Phe Leu Lys Leu Tyr Glu Lys Gly Leu Ala Tyr Met Asp Glu Val Pro
145 150 155 160
Val Asn Trp Cys Pro Ala Leu Gly Thr Val Leu Ala Asn Glu Glu Val
165 170 175
Ile Asn Gly Arg Ser Glu Arg Gly Gly His Pro Val Ile Arg Lys Pro
180 185 190
Met Arg Gln Trp Met Leu Lys Ile Thr Ala Tyr Ala Asp Arg Leu Leu
195 200 205
Glu Asp Leu Glu Glu Leu Asp Trp Pro Glu Ser Ile Lys Glu Met Gln
210 215 220
Arg Asn Trp Ile Gly Arg Ser Glu Gly Ala Glu Ile Glu Phe Ala Val
225 230 235 240
Asp Gly His Asp Glu Ser Phe Thr Val Phe Thr Thr Arg Pro Asp Thr
245 250 255
Leu Phe Gly Ala Thr Tyr Ala Val Leu Ala Pro Glu His Pro Leu Val
260 265 270
Glu Lys Ile Thr Thr Pro Glu Gln Lys Pro Ala Val Asp Ala Tyr Leu
275 280 285
Lys Glu Val Gln Ser Lys Ser Asp Leu Glu Arg Thr Asp Leu Ala Lys
290 295 300
Glu Lys Thr Gly Val Phe Thr Gly Ala Tyr Ala Ile His Pro Val Thr
305 310 315 320
Gly Asp Lys Leu Pro Ile Trp Ile Ala Asp Tyr Val Leu Met Gly Tyr
325 330 335
Gly Thr Gly Ala Ile Met Ala Val Pro Ala His Asp Glu Arg Asp Tyr
340 345 350
Glu Phe Ala Lys Thr Phe Asn Leu Pro Ile Lys Glu Val Val Ala Gly
355 360 365
Gly Asn Val Glu Asn Glu Pro Tyr Thr Gly Asp Gly Glu His Ile Asn
370 375 380
Ser Glu Phe Leu Asn Gly Leu Asn Lys Gln Glu Ala Ile Glu Lys Met
385 390 395 400
Ile Ala Trp Leu Glu Glu Asn Gly Lys Gly Gln Lys Lys Val Ser Tyr
405 410 415
Arg Leu Arg Asp Trp Leu Phe Ser Arg Gln Arg Tyr Trp Gly Glu Pro
420 425 430
Ile Pro Val Ile His Trp Glu Asp Gly Thr Met Thr Thr Val Pro Glu
435 440 445
Glu Glu Leu Pro Leu Val Leu Pro Lys Thr Asp Glu Ile Lys Pro Ser
450 455 460
Gly Thr Gly Glu Ser Pro Leu Ala Asn Ile Glu Glu Trp Val Asn Val
465 470 475 480
Val Asp Pro Lys Thr Gly Lys Lys Gly Arg Arg Glu Thr Asn Thr Met
485 490 495
Pro Gln Trp Ala Gly Ser Cys Trp Tyr Tyr Leu Arg Tyr Ile Asp Pro
500 505 510
His Asn Asp Lys Gln Leu Ala Asp Pro Glu Lys Leu Lys Gln Trp Leu
515 520 525
Pro Val Asp Val Tyr Ile Gly Gly Ala Glu His Ala Val Leu His Leu
530 535 540
Leu Tyr Ala Arg Phe Trp His Lys Val Leu Tyr Asp Leu Gly Ile Val
545 550 555 560
Pro Thr Lys Glu Pro Phe Gln Lys Leu Phe Asn Gln Gly Met Ile Leu
565 570 575
Gly Glu Asn Asn Glu Lys Met Ser Lys Ser Lys Gly Asn Val Val Asn
580 585 590
Pro Asp Asp Ile Val Glu Ser His Gly Ala Asp Thr Leu Arg Leu Tyr
595 600 605
Glu Met Phe Met Gly Pro Leu Glu Ala Ser Ile Ala Trp Ser Thr Lys
610 615 620
Gly Leu Asp Gly Ala Arg Arg Phe Leu Glu Arg Val Trp Arg Leu Phe
625 630 635 640
Val Thr Glu Asp Gly Gln Leu Asn Pro Asn Ile Val Asp Glu Pro Ala
645 650 655
Asn Asp Thr Leu Glu Arg Val Tyr His Gln Thr Val Lys Lys Val Thr
660 665 670
Glu Asp Tyr Glu Ala Leu Arg Phe Asn Thr Ala Ile Ser Gln Leu Met
675 680 685
Val Phe Ile Asn Glu Ala Tyr Lys Ala Glu Gln Met Lys Lys Glu Tyr
690 695 700
Met Glu Gly Phe Val Lys Leu Leu Ser Pro Val Cys Pro His Ile Gly
705 710 715 720
Glu Glu Leu Trp Gln Lys Leu Gly His Thr Asp Thr Ile Ala Tyr Glu
725 730 735
Pro Trp Pro Thr Tyr Asp Glu Ala Lys Leu Val Glu Asp Val Val Glu
740 745 750
Ile Val Ile Gln Ile Asn Gly Lys Val Arg Ala Lys Leu Asn Val Pro
755 760 765
Ala Asp Leu Ser Lys Glu Ala Leu Glu Glu Arg Ala Leu Ala Asp Glu
770 775 780
Lys Ile Lys Glu Gln Leu Ala Gly Lys Thr Val Arg Lys Val Ile Thr
785 790 795 800
Val Pro Gly Lys Leu Val Asn Ile Val Ala Asn
805 810
<210> 111
<211> 1482
<212> DNA
<213> Geobacillus subterraneus
<400> 111
atgagccatg aagaattgaa cgaccaattg cgtgtccgcc gggaaaagtt aaaaaaaatc 60
gaagagctag gtgtcgaccc gtttggcaaa cggttcgagc gcacgcataa agcagaagag 120
ctgtttaaac tgtacggcga tttgtccaaa gaagaacttg aagatcagca aattgaagtc 180
gctgtcgccg gccgcattat gacgaaacgc ggtaaaggaa aagcaggatt tgctcacatt 240
caagacgtca cagggcaaat tcaaatttat gtccgccaag acgatgtcgg tgaacagcaa 300
tatgagctgt ttaaaatctc tgaccttggt gatatcgtcg gtgtgcgcgg cactatgttc 360
aaaacaaaag tcggcgagct ttccatcaaa gtgtcatcat atgaattttt aacaaaagca 420
ttgcgtccat tgccggaaaa ataccatggt ttaaaggacg tcgaacaacg ttaccgccaa 480
cgttatctcg acttaactat gaatccgcaa agtaagcaga cgtttatcac ccgtagtctc 540
attattcaat cgatgcggcg ttatctcgac agccaaggtt atttggaagt cgaaacaccg 600
atgatgcacg ccatagcagg tggtgcggct gcacgtccgt ttattacgca ccataatgcc 660
cttgatatga cactttatat gcgaatcgcc atcgaactcc atttaaaacg gctcatcgtc 720
ggcggtttgg aaaaagtgta tgaaatcgga cgcgtcttcc ggaatgaggg gatttccacc 780
cgtcacaatc cggagtttac gatgcttgaa ctgtacgagg catatgccga cttccgtgac 840
atcatgaaat tgacagaaaa cttaattgct cacattgcca cggaagtgct tggcacgacg 900
aaaattcaat acggcgaaca taccgtcgat ttaacgcctg aatggcggcg acttcatatg 960
gtcgatgcga ttaaagaata cgtcggcgtt gatttctggc ggcacatgga cgacgaggaa 1020
gcgcgggcgt tggcgaaaga acatggggtc gaaatcgccc cgcacatgac gtttggtcat 1080
atcgtcaatg aattttttga acaaaaagtc gagtcgcaac tcatccaacc gacgttcatt 1140
tatggccacc ctgtcgaaat ttcgccgtta gctaagaaaa acccggacga tccacgcttt 1200
accgatcgat ttgagctatt tatcgttgga cgtgaacatg cgaacgcgtt tacggaacta 1260
aacgatccga tcgaccagcg ccaacgtttc gaagcacagt tgaaagaacg tgaacaaggg 1320
aacgatgaag cgcacgaaat ggacgaagat ttcctcgaag cgctcgagta cggtatgcct 1380
ccaacaggcg gactcggcat cggcgttgac cgtctagtca tgctcttgac taactctccg 1440
tccattcggg atgtgttact cttcccgcaa atgcgtcata aa 1482
<210> 112
<211> 494
<212> PRT
<213> Geobacillus subterraneus
<400> 112
Met Ser His Glu Glu Leu Asn Asp Gln Leu Arg Val Arg Arg Glu Lys
1 5 10 15
Leu Lys Lys Ile Glu Glu Leu Gly Val Asp Pro Phe Gly Lys Arg Phe
20 25 30
Glu Arg Thr His Lys Ala Glu Glu Leu Phe Lys Leu Tyr Gly Asp Leu
35 40 45
Ser Lys Glu Glu Leu Glu Asp Gln Gln Ile Glu Val Ala Val Ala Gly
50 55 60
Arg Ile Met Thr Lys Arg Gly Lys Gly Lys Ala Gly Phe Ala His Ile
65 70 75 80
Gln Asp Val Thr Gly Gln Ile Gln Ile Tyr Val Arg Gln Asp Asp Val
85 90 95
Gly Glu Gln Gln Tyr Glu Leu Phe Lys Ile Ser Asp Leu Gly Asp Ile
100 105 110
Val Gly Val Arg Gly Thr Met Phe Lys Thr Lys Val Gly Glu Leu Ser
115 120 125
Ile Lys Val Ser Ser Tyr Glu Phe Leu Thr Lys Ala Leu Arg Pro Leu
130 135 140
Pro Glu Lys Tyr His Gly Leu Lys Asp Val Glu Gln Arg Tyr Arg Gln
145 150 155 160
Arg Tyr Leu Asp Leu Thr Met Asn Pro Gln Ser Lys Gln Thr Phe Ile
165 170 175
Thr Arg Ser Leu Ile Ile Gln Ser Met Arg Arg Tyr Leu Asp Ser Gln
180 185 190
Gly Tyr Leu Glu Val Glu Thr Pro Met Met His Ala Ile Ala Gly Gly
195 200 205
Ala Ala Ala Arg Pro Phe Ile Thr His His Asn Ala Leu Asp Met Thr
210 215 220
Leu Tyr Met Arg Ile Ala Ile Glu Leu His Leu Lys Arg Leu Ile Val
225 230 235 240
Gly Gly Leu Glu Lys Val Tyr Glu Ile Gly Arg Val Phe Arg Asn Glu
245 250 255
Gly Ile Ser Thr Arg His Asn Pro Glu Phe Thr Met Leu Glu Leu Tyr
260 265 270
Glu Ala Tyr Ala Asp Phe Arg Asp Ile Met Lys Leu Thr Glu Asn Leu
275 280 285
Ile Ala His Ile Ala Thr Glu Val Leu Gly Thr Thr Lys Ile Gln Tyr
290 295 300
Gly Glu His Thr Val Asp Leu Thr Pro Glu Trp Arg Arg Leu His Met
305 310 315 320
Val Asp Ala Ile Lys Glu Tyr Val Gly Val Asp Phe Trp Arg His Met
325 330 335
Asp Asp Glu Glu Ala Arg Ala Leu Ala Lys Glu His Gly Val Glu Ile
340 345 350
Ala Pro His Met Thr Phe Gly His Ile Val Asn Glu Phe Phe Glu Gln
355 360 365
Lys Val Glu Ser Gln Leu Ile Gln Pro Thr Phe Ile Tyr Gly His Pro
370 375 380
Val Glu Ile Ser Pro Leu Ala Lys Lys Asn Pro Asp Asp Pro Arg Phe
385 390 395 400
Thr Asp Arg Phe Glu Leu Phe Ile Val Gly Arg Glu His Ala Asn Ala
405 410 415
Phe Thr Glu Leu Asn Asp Pro Ile Asp Gln Arg Gln Arg Phe Glu Ala
420 425 430
Gln Leu Lys Glu Arg Glu Gln Gly Asn Asp Glu Ala His Glu Met Asp
435 440 445
Glu Asp Phe Leu Glu Ala Leu Glu Tyr Gly Met Pro Pro Thr Gly Gly
450 455 460
Leu Gly Ile Gly Val Asp Arg Leu Val Met Leu Leu Thr Asn Ser Pro
465 470 475 480
Ser Ile Arg Asp Val Leu Leu Phe Pro Gln Met Arg His Lys
485 490
<210> 113
<211> 1950
<212> DNA
<213> Geobacillus subterraneus
<400> 113
atggagaaaa agacgtttta tttgacgacg ccgatttatt atccgagcga caaattgcac 60
atcggccatg cttatacaac agtggcgggg gatacgctag cgcgctataa acggatgcgc 120
ggttacgatg ttatgtattt gacgggaacc gatgagcacg ggcaaaaaat tcaacgcaag 180
gcggaggaaa aaggagtaac gccgcagcaa tatgtcgatg agatcgtcgc tggcattcag 240
gagctatgga aaaagctcga catttcttat gacgatttca tccgtacaac gcaggagcgg 300
cataaaaaag tagtcgaaaa gattttcgcg cgtcttgtcg aacaagggga tatttattta 360
ggtgaatatg aaggatggta ttgcacgcca tgcgaatcgt tttacactga gcgacagctt 420
gtcgacggca actgcccgga ctgtggtcgt ccggttgaaa aagtgaaaga gcagtcgtac 480
tttttccgaa tgagcaaata cgtcgaccgt ttgcttcaat attatgagga aaatccagat 540
ttcatccagc cggaatcgcg gaaaaacgaa atgattaaca attttattaa gccggggctt 600
gaagatttag ctgtgtcgcg gacgacgttt gactggggca ttaaagtgcc gggcgatccg 660
aaacatgtca tttacgtctg gattgacgcg cttgccaact atattacagc gctcggttac 720
ggcacggaca atgatgaaaa gttccgcaaa tattggccgg ccgatgtcca tttagtcggc 780
aaggaaatca tccgctttca tacgatttat tggccgatta tgctcatggc gcttgacttg 840
ccgctgccga aaaaagtatt cggtcatggc tggctgctca tgaaagacgg gaaaatgtcg 900
aaatcgaaag gcaatgtcgt tgacccggtg acgttgatcg atcgatacgg actcgatgcg 960
cttcgttatt atttactcag ggaagtgccg ttcggttctg acggcgtatt cacgccggaa 1020
ggatttattg agcgcatcaa ctacgattta gccaatgacc taggcaattt attgaatcgt 1080
acagtagcga tgattaagaa atattttgat ggggtgattc cgccgtaccg cggtccgaaa 1140
acgccgtttg acgaagagct ggtacaaacg gcgcgtgagg tggtccgtca gtatgaggaa 1200
gcgatggaac ggatggagtt ttccgttgcc cttgcttcgg tttggcaact gattggccgg 1260
acgaacaaat acattgatga gacgcagcca tgggtattgg ccaaagatga aagcaaacgg 1320
gaagagcttg cttctgtcat gacccaccta gccgagtcgc tccgccatac ggcagtgctg 1380
ttgcagccgt ttttgacacg cacgccagag cgcatttttg cccagctcgg cattgccgac 1440
cgttcattaa aagagtggga tagcttgtac gagttcgggc tcattccgga aggaacaaac 1500
gtgcaaaaag gagaaccact gttcccgcgc cttgatattg aagcggaagt cgagtacatt 1560
aaggcgcata tgcaaggcgg caagccggcg gtggaacccg ttaaagagga gaagcaagcg 1620
gctgagacgg ccgaaatctc aattgatgag tttgccaaag ttgacttgcg cgttgctgaa 1680
gtcgtgcatg ctgaacggat gaaaaacgcc aataagctgt tgaagctcca acttgatctt 1740
ggcggcgaga aacggcaagt catctctggt atcgctgaat tttacaaacc agaggaactc 1800
atcggcaaaa aggtcatttg cgtcgccaat ttaaaaccgg ccaaactgcg cggtgagtgg 1860
tcggaaggaa tgattttggc cggcggtaac ggcggagagt tttcactggc gaccgtcgat 1920
caacatgtgc caaacggaac aaaaattaaa 1950
<210> 114
<211> 650
<212> PRT
<213> Geobacillus subterraneus
<400> 114
Met Glu Lys Lys Thr Phe Tyr Leu Thr Thr Pro Ile Tyr Tyr Pro Ser
1 5 10 15
Asp Lys Leu His Ile Gly His Ala Tyr Thr Thr Val Ala Gly Asp Thr
20 25 30
Leu Ala Arg Tyr Lys Arg Met Arg Gly Tyr Asp Val Met Tyr Leu Thr
35 40 45
Gly Thr Asp Glu His Gly Gln Lys Ile Gln Arg Lys Ala Glu Glu Lys
50 55 60
Gly Val Thr Pro Gln Gln Tyr Val Asp Glu Ile Val Ala Gly Ile Gln
65 70 75 80
Glu Leu Trp Lys Lys Leu Asp Ile Ser Tyr Asp Asp Phe Ile Arg Thr
85 90 95
Thr Gln Glu Arg His Lys Lys Val Val Glu Lys Ile Phe Ala Arg Leu
100 105 110
Val Glu Gln Gly Asp Ile Tyr Leu Gly Glu Tyr Glu Gly Trp Tyr Cys
115 120 125
Thr Pro Cys Glu Ser Phe Tyr Thr Glu Arg Gln Leu Val Asp Gly Asn
130 135 140
Cys Pro Asp Cys Gly Arg Pro Val Glu Lys Val Lys Glu Gln Ser Tyr
145 150 155 160
Phe Phe Arg Met Ser Lys Tyr Val Asp Arg Leu Leu Gln Tyr Tyr Glu
165 170 175
Glu Asn Pro Asp Phe Ile Gln Pro Glu Ser Arg Lys Asn Glu Met Ile
180 185 190
Asn Asn Phe Ile Lys Pro Gly Leu Glu Asp Leu Ala Val Ser Arg Thr
195 200 205
Thr Phe Asp Trp Gly Ile Lys Val Pro Gly Asp Pro Lys His Val Ile
210 215 220
Tyr Val Trp Ile Asp Ala Leu Ala Asn Tyr Ile Thr Ala Leu Gly Tyr
225 230 235 240
Gly Thr Asp Asn Asp Glu Lys Phe Arg Lys Tyr Trp Pro Ala Asp Val
245 250 255
His Leu Val Gly Lys Glu Ile Ile Arg Phe His Thr Ile Tyr Trp Pro
260 265 270
Ile Met Leu Met Ala Leu Asp Leu Pro Leu Pro Lys Lys Val Phe Gly
275 280 285
His Gly Trp Leu Leu Met Lys Asp Gly Lys Met Ser Lys Ser Lys Gly
290 295 300
Asn Val Val Asp Pro Val Thr Leu Ile Asp Arg Tyr Gly Leu Asp Ala
305 310 315 320
Leu Arg Tyr Tyr Leu Leu Arg Glu Val Pro Phe Gly Ser Asp Gly Val
325 330 335
Phe Thr Pro Glu Gly Phe Ile Glu Arg Ile Asn Tyr Asp Leu Ala Asn
340 345 350
Asp Leu Gly Asn Leu Leu Asn Arg Thr Val Ala Met Ile Lys Lys Tyr
355 360 365
Phe Asp Gly Val Ile Pro Pro Tyr Arg Gly Pro Lys Thr Pro Phe Asp
370 375 380
Glu Glu Leu Val Gln Thr Ala Arg Glu Val Val Arg Gln Tyr Glu Glu
385 390 395 400
Ala Met Glu Arg Met Glu Phe Ser Val Ala Leu Ala Ser Val Trp Gln
405 410 415
Leu Ile Gly Arg Thr Asn Lys Tyr Ile Asp Glu Thr Gln Pro Trp Val
420 425 430
Leu Ala Lys Asp Glu Ser Lys Arg Glu Glu Leu Ala Ser Val Met Thr
435 440 445
His Leu Ala Glu Ser Leu Arg His Thr Ala Val Leu Leu Gln Pro Phe
450 455 460
Leu Thr Arg Thr Pro Glu Arg Ile Phe Ala Gln Leu Gly Ile Ala Asp
465 470 475 480
Arg Ser Leu Lys Glu Trp Asp Ser Leu Tyr Glu Phe Gly Leu Ile Pro
485 490 495
Glu Gly Thr Asn Val Gln Lys Gly Glu Pro Leu Phe Pro Arg Leu Asp
500 505 510
Ile Glu Ala Glu Val Glu Tyr Ile Lys Ala His Met Gln Gly Gly Lys
515 520 525
Pro Ala Val Glu Pro Val Lys Glu Glu Lys Gln Ala Ala Glu Thr Ala
530 535 540
Glu Ile Ser Ile Asp Glu Phe Ala Lys Val Asp Leu Arg Val Ala Glu
545 550 555 560
Val Val His Ala Glu Arg Met Lys Asn Ala Asn Lys Leu Leu Lys Leu
565 570 575
Gln Leu Asp Leu Gly Gly Glu Lys Arg Gln Val Ile Ser Gly Ile Ala
580 585 590
Glu Phe Tyr Lys Pro Glu Glu Leu Ile Gly Lys Lys Val Ile Cys Val
595 600 605
Ala Asn Leu Lys Pro Ala Lys Leu Arg Gly Glu Trp Ser Glu Gly Met
610 615 620
Ile Leu Ala Gly Gly Asn Gly Gly Glu Phe Ser Leu Ala Thr Val Asp
625 630 635 640
Gln His Val Pro Asn Gly Thr Lys Ile Lys
645 650
<210> 115
<211> 1074
<212> DNA
<213> Geobacillus subterraneus
<400> 115
atgagggacg ggttttttta ttttgttaga ggagggattg gcgtgaaaga acggttgcat 60
gagcttgaac gagaagcgct tgaaaaaatt gaacaagctg gcgatttaaa agcgctcaac 120
gatgtgcgtg tcgcctattt aggcaaaaaa gggccgatta ccgaagtgct gcgcggcatg 180
ggagcattgc cgtcagaaga gcgtccgaaa attggtgcgc ttgccaatga ggtaagagag 240
gcgatccaaa aggcgctcga agcaaaacaa acgaaactgg aagaagaaga agtcgagcgg 300
aagttggcgg ctgaagcgat cgatgtgacg cttccgggcc gtccggtgaa actggggaat 360
cctcatccgc tgacgcgcgt catcgaggaa attgaagatt tgtttatcgg catgggctat 420
acggtcgccg aaggtccgga agtcgagacc gattattaca attttgaggc gctcaatttg 480
ccgaaaggac acccggcccg cgatatgcaa gattcgtttt atattacgga agaaattctg 540
cttcgcaccc acacgtcgcc gatgcaggca cggacgatgg aaaaacatcg cgggcgcggt 600
ccggtaaaaa tcatttgccc ggggaaagtg tatcgccgcg ataccgatga tgcgacccat 660
tcacatcagt ttacgcaaat tgaaggattg gttgttgacc gcaacatccg gatgagcgat 720
ttaaaaggga cgctgcgcga atttgcccgc aagctgttcg gtgaagggcg cgacatccgt 780
tttcgtccga gctttttccc gtttaccgag ccttcagtcg aggtcgatgt gtcctgcttc 840
cgctgcgaag ggcacggctg cagcgtttgc aaaggtacgg gctggattga aattttaggc 900
gctggcatgg tgcacccgaa cgtgcttgag atggccggct ttgattcgaa aacgtatacc 960
ggatttgcgt tcggcatggg gccggagcgg atcgcgatgt tgaaatacgg cattgatgac 1020
atccgccatt tctatcagaa cgatcttcgt ttcttgcaac aatttttgcg tgtc 1074
<210> 116
<211> 358
<212> PRT
<213> Geobacillus subterraneus
<400> 116
Met Arg Asp Gly Phe Phe Tyr Phe Val Arg Gly Gly Ile Gly Val Lys
1 5 10 15
Glu Arg Leu His Glu Leu Glu Arg Glu Ala Leu Glu Lys Ile Glu Gln
20 25 30
Ala Gly Asp Leu Lys Ala Leu Asn Asp Val Arg Val Ala Tyr Leu Gly
35 40 45
Lys Lys Gly Pro Ile Thr Glu Val Leu Arg Gly Met Gly Ala Leu Pro
50 55 60
Ser Glu Glu Arg Pro Lys Ile Gly Ala Leu Ala Asn Glu Val Arg Glu
65 70 75 80
Ala Ile Gln Lys Ala Leu Glu Ala Lys Gln Thr Lys Leu Glu Glu Glu
85 90 95
Glu Val Glu Arg Lys Leu Ala Ala Glu Ala Ile Asp Val Thr Leu Pro
100 105 110
Gly Arg Pro Val Lys Leu Gly Asn Pro His Pro Leu Thr Arg Val Ile
115 120 125
Glu Glu Ile Glu Asp Leu Phe Ile Gly Met Gly Tyr Thr Val Ala Glu
130 135 140
Gly Pro Glu Val Glu Thr Asp Tyr Tyr Asn Phe Glu Ala Leu Asn Leu
145 150 155 160
Pro Lys Gly His Pro Ala Arg Asp Met Gln Asp Ser Phe Tyr Ile Thr
165 170 175
Glu Glu Ile Leu Leu Arg Thr His Thr Ser Pro Met Gln Ala Arg Thr
180 185 190
Met Glu Lys His Arg Gly Arg Gly Pro Val Lys Ile Ile Cys Pro Gly
195 200 205
Lys Val Tyr Arg Arg Asp Thr Asp Asp Ala Thr His Ser His Gln Phe
210 215 220
Thr Gln Ile Glu Gly Leu Val Val Asp Arg Asn Ile Arg Met Ser Asp
225 230 235 240
Leu Lys Gly Thr Leu Arg Glu Phe Ala Arg Lys Leu Phe Gly Glu Gly
245 250 255
Arg Asp Ile Arg Phe Arg Pro Ser Phe Phe Pro Phe Thr Glu Pro Ser
260 265 270
Val Glu Val Asp Val Ser Cys Phe Arg Cys Glu Gly His Gly Cys Ser
275 280 285
Val Cys Lys Gly Thr Gly Trp Ile Glu Ile Leu Gly Ala Gly Met Val
290 295 300
His Pro Asn Val Leu Glu Met Ala Gly Phe Asp Ser Lys Thr Tyr Thr
305 310 315 320
Gly Phe Ala Phe Gly Met Gly Pro Glu Arg Ile Ala Met Leu Lys Tyr
325 330 335
Gly Ile Asp Asp Ile Arg His Phe Tyr Gln Asn Asp Leu Arg Phe Leu
340 345 350
Gln Gln Phe Leu Arg Val
355
<210> 117
<211> 2412
<212> DNA
<213> Geobacillus subterraneus
<400> 117
atgctcgttt cttatcgttg gctaggcgaa tacgtcgatt tgacgggcgt gacggcggaa 60
caactcgctg atcgcattac aaaaagcggc attgaagtcg agcgggttga agcgcttgag 120
cggggaatga aaggagtcgt catcggccat gtgctcgaat gcgagccaca cccaaacgcc 180
gataaactgc ggaaatgtct tgttgatctt ggcgaaggag agccggtgca aatcatttgc 240
ggtgccccga acgtcgccaa ggggcaaaaa gttgctgtag cgaaagttgg agcgagactg 300
ccgggcaatt ttaaaatcaa acgggcgaag ctgcgcggcg aagagtcgaa cggcatgatt 360
tgctcgctcc aagaactcgg tgttgaaaca aaagtcgtgc cgaaagaata cgccgaaggc 420
attttcgtct tcccaagcga cgcgccggtc ggcgctgatg cgcttgaatg gctcggcttg 480
cacgatgaag tgctcgaact cgccttgacg ccgaatcgcg ccgattgctt aagcatgctt 540
ggcgttgcct acgaagtcgc tgcgattctc ggccgcgatg tgaagttgcc ggaaacggcg 600
gtgaacgaaa atgaagaaag cgtccatgac tacatttctg tccgtgtcga ggcgccggaa 660
gacaatccgc tgtacgccgg acggatcgtg aaaaacgtcc aaatcggccc gtcgccgctt 720
tggatgcaag cgcgcttgat ggcggccggc attcgtccac acaacaatgt tgtcgatatc 780
accaactaca ttttgcttga gtacggccag ccgcttcacg cgtttgacta cgaccgtctc 840
ggttcgaagg agatcgtcgt tcgtcgtgcc aaggcgggag aaatgatcgt gacgcttgac 900
gatgtcgagc ggaagctgac tgaagatcat ctcgtcatca caaacggccg tgagccggtc 960
gccttagccg gtgtgatggg cggagcgaac tcggaagtgc aggatgacac gaaaacagtg 1020
ttcatcgaag ccgcgtattt tacgagcccg gtcatccgcc aggcggtgaa agaccacggg 1080
ttgcgcagcg aagcgagcac ccggtttgaa aaagggattg atccggcgcg gacgaaagaa 1140
gcgctcgagc gcgctgctgc tttgatggca gaatacgccg gcggcgaggt cgtcagcggt 1200
atcgtggaag ctaatacatg gaaagaagag ccggttgtcg taacggtggc gctggaacgc 1260
atcaacggcg tcctcggcac agcgatgacg aaagaggaag tagctggcat tctttcaaac 1320
ttgcaattct cgtttacgga agataatgga acgtttacaa tccatgttcc atcgcgccgc 1380
cgcgatatta cgatcgaaga agatattatc gaggaagtcg cccgtttgta tggctacgac 1440
catttgccag cgactttgcc ggtggccgaa gcaaaaccgg gcgagttgac accgtaccaa 1500
gcgaaacgcc gccgtgtccg ccgctatttc gaaggcgcgg gcttgttcca ggcgatcacg 1560
tattcgctta ccagtccgga caaagcgacg cggtttgctt tggagacaac cgaaccagtc 1620
cgcttggcgt tgccgatgag tgaggagcgg agcgttctcc ggcaaagctt ggtgccgcat 1680
ttgctcgaag cggcgagcta caaccgtgcc cgccaagttg agaacgtcgc gctatatgaa 1740
atcggctctg tctatttgtc caagggggaa aatgtccaac cggcggaaaa agaacggctc 1800
gccggcgtca tcaccggttt atggcatgcc cacctttggc aaggagagaa aaaagcagct 1860
gatttctatg ttgcaaaagg cgtgcttgac ggcttgttcg ccctgcttgg gctgtctgat 1920
cgcatcagct accgtccggc gaagcgtgct gatttgcatc tggggcggac agcggagatt 1980
gtgcttgacg gcaaagagat cggctttgtc ggccagctcc atccggctgt acaaaaagag 2040
tacgatttga aagaaacgta tgtctttgaa ctcgccttcg ctgagctact gaatacagaa 2100
ggcgaaacga tccgttacga gtcgattccg cgcttcccgt cagtcgtgcg cgacatcgct 2160
ttagtcgtcg acgacaatgt cgaagcaggt gctctcaagc aggcgatcgc cgaagcgggg 2220
aacccgctat taaaagacgt ggccctcttt gacgtctata aaggcgaccg tctgccggcc 2280
gggaaaaaat cgctcgcctt ctcgctccgc tactacgatc cggaacggac gctcactgat 2340
gaggaagtta ctgccgtcca tgaacgggtt ttggcagcgg tcgaggagca gtttggcgcg 2400
gtgttgcgcg gg 2412
<210> 118
<211> 804
<212> PRT
<213> Geobacillus subterraneus
<400> 118
Met Leu Val Ser Tyr Arg Trp Leu Gly Glu Tyr Val Asp Leu Thr Gly
1 5 10 15
Val Thr Ala Glu Gln Leu Ala Asp Arg Ile Thr Lys Ser Gly Ile Glu
20 25 30
Val Glu Arg Val Glu Ala Leu Glu Arg Gly Met Lys Gly Val Val Ile
35 40 45
Gly His Val Leu Glu Cys Glu Pro His Pro Asn Ala Asp Lys Leu Arg
50 55 60
Lys Cys Leu Val Asp Leu Gly Glu Gly Glu Pro Val Gln Ile Ile Cys
65 70 75 80
Gly Ala Pro Asn Val Ala Lys Gly Gln Lys Val Ala Val Ala Lys Val
85 90 95
Gly Ala Arg Leu Pro Gly Asn Phe Lys Ile Lys Arg Ala Lys Leu Arg
100 105 110
Gly Glu Glu Ser Asn Gly Met Ile Cys Ser Leu Gln Glu Leu Gly Val
115 120 125
Glu Thr Lys Val Val Pro Lys Glu Tyr Ala Glu Gly Ile Phe Val Phe
130 135 140
Pro Ser Asp Ala Pro Val Gly Ala Asp Ala Leu Glu Trp Leu Gly Leu
145 150 155 160
His Asp Glu Val Leu Glu Leu Ala Leu Thr Pro Asn Arg Ala Asp Cys
165 170 175
Leu Ser Met Leu Gly Val Ala Tyr Glu Val Ala Ala Ile Leu Gly Arg
180 185 190
Asp Val Lys Leu Pro Glu Thr Ala Val Asn Glu Asn Glu Glu Ser Val
195 200 205
His Asp Tyr Ile Ser Val Arg Val Glu Ala Pro Glu Asp Asn Pro Leu
210 215 220
Tyr Ala Gly Arg Ile Val Lys Asn Val Gln Ile Gly Pro Ser Pro Leu
225 230 235 240
Trp Met Gln Ala Arg Leu Met Ala Ala Gly Ile Arg Pro His Asn Asn
245 250 255
Val Val Asp Ile Thr Asn Tyr Ile Leu Leu Glu Tyr Gly Gln Pro Leu
260 265 270
His Ala Phe Asp Tyr Asp Arg Leu Gly Ser Lys Glu Ile Val Val Arg
275 280 285
Arg Ala Lys Ala Gly Glu Met Ile Val Thr Leu Asp Asp Val Glu Arg
290 295 300
Lys Leu Thr Glu Asp His Leu Val Ile Thr Asn Gly Arg Glu Pro Val
305 310 315 320
Ala Leu Ala Gly Val Met Gly Gly Ala Asn Ser Glu Val Gln Asp Asp
325 330 335
Thr Lys Thr Val Phe Ile Glu Ala Ala Tyr Phe Thr Ser Pro Val Ile
340 345 350
Arg Gln Ala Val Lys Asp His Gly Leu Arg Ser Glu Ala Ser Thr Arg
355 360 365
Phe Glu Lys Gly Ile Asp Pro Ala Arg Thr Lys Glu Ala Leu Glu Arg
370 375 380
Ala Ala Ala Leu Met Ala Glu Tyr Ala Gly Gly Glu Val Val Ser Gly
385 390 395 400
Ile Val Glu Ala Asn Thr Trp Lys Glu Glu Pro Val Val Val Thr Val
405 410 415
Ala Leu Glu Arg Ile Asn Gly Val Leu Gly Thr Ala Met Thr Lys Glu
420 425 430
Glu Val Ala Gly Ile Leu Ser Asn Leu Gln Phe Ser Phe Thr Glu Asp
435 440 445
Asn Gly Thr Phe Thr Ile His Val Pro Ser Arg Arg Arg Asp Ile Thr
450 455 460
Ile Glu Glu Asp Ile Ile Glu Glu Val Ala Arg Leu Tyr Gly Tyr Asp
465 470 475 480
His Leu Pro Ala Thr Leu Pro Val Ala Glu Ala Lys Pro Gly Glu Leu
485 490 495
Thr Pro Tyr Gln Ala Lys Arg Arg Arg Val Arg Arg Tyr Phe Glu Gly
500 505 510
Ala Gly Leu Phe Gln Ala Ile Thr Tyr Ser Leu Thr Ser Pro Asp Lys
515 520 525
Ala Thr Arg Phe Ala Leu Glu Thr Thr Glu Pro Val Arg Leu Ala Leu
530 535 540
Pro Met Ser Glu Glu Arg Ser Val Leu Arg Gln Ser Leu Val Pro His
545 550 555 560
Leu Leu Glu Ala Ala Ser Tyr Asn Arg Ala Arg Gln Val Glu Asn Val
565 570 575
Ala Leu Tyr Glu Ile Gly Ser Val Tyr Leu Ser Lys Gly Glu Asn Val
580 585 590
Gln Pro Ala Glu Lys Glu Arg Leu Ala Gly Val Ile Thr Gly Leu Trp
595 600 605
His Ala His Leu Trp Gln Gly Glu Lys Lys Ala Ala Asp Phe Tyr Val
610 615 620
Ala Lys Gly Val Leu Asp Gly Leu Phe Ala Leu Leu Gly Leu Ser Asp
625 630 635 640
Arg Ile Ser Tyr Arg Pro Ala Lys Arg Ala Asp Leu His Leu Gly Arg
645 650 655
Thr Ala Glu Ile Val Leu Asp Gly Lys Glu Ile Gly Phe Val Gly Gln
660 665 670
Leu His Pro Ala Val Gln Lys Glu Tyr Asp Leu Lys Glu Thr Tyr Val
675 680 685
Phe Glu Leu Ala Phe Ala Glu Leu Leu Asn Thr Glu Gly Glu Thr Ile
690 695 700
Arg Tyr Glu Ser Ile Pro Arg Phe Pro Ser Val Val Arg Asp Ile Ala
705 710 715 720
Leu Val Val Asp Asp Asn Val Glu Ala Gly Ala Leu Lys Gln Ala Ile
725 730 735
Ala Glu Ala Gly Asn Pro Leu Leu Lys Asp Val Ala Leu Phe Asp Val
740 745 750
Tyr Lys Gly Asp Arg Leu Pro Ala Gly Lys Lys Ser Leu Ala Phe Ser
755 760 765
Leu Arg Tyr Tyr Asp Pro Glu Arg Thr Leu Thr Asp Glu Glu Val Thr
770 775 780
Ala Val His Glu Arg Val Leu Ala Ala Val Glu Glu Gln Phe Gly Ala
785 790 795 800
Val Leu Arg Gly
<210> 119
<211> 1755
<212> DNA
<213> Geobacillus subterraneus
<400> 119
atgacattca aaaattcttc ctataatgaa agagagaaaa cgaggtggct attgatgaga 60
caaagtcaag ggtttattcc gacattgcgc gaagtgccgg cggacgcgga agtgaaaagc 120
catcagctcc tgttgcgggc cggcttcgtc cgccaaagcg caagcggcgt ctacacgttt 180
ttgccgctcg ggcaacgtgt tttgcaaaaa gtggaagcga ttattcgtga ggagatgaat 240
cgcgccggag cattggagct tctcatgcct gctttgcagc cggctgagct ttggcagcag 300
tccgggcgct ggtattcgta tggaccggag ctcatgcgcc tgaaagaccg tcacgagcgc 360
gatttcgttc tcggaccgac acacgaagag atgattacta cgatcgttcg cgatgaagtg 420
aaaacgtata agcggctgcc gcttatcttg tatcaaattc aaacgaaatt ccgtgatgaa 480
aaacgtccgc gtttcgggct gttgcgcggt cgcgagttca tcatgaaaga tgcgtattca 540
ttccacacat cgcaggaaag tttggacgaa acgtacaata aaatgtatga agcgtacgcg 600
aacattttcc gccgctgcgg cttaaatttc cgcgctgtca ttgctgactc cggagcgatg 660
ggcggcaaag atacgcacga gtttatggtg ctgtctgata ttggcgagga tacgatcgct 720
tattccgatg cgtccgacta tgcggccaac attgaaatgg caccggtcgt cactacgtat 780
gaaaaaagca gtgagccgct ggtggaactg aaaaaagtgg cgaccccgga gcaaaaaacg 840
attgctgaag ttgcttcgta tttgcaagta gcaccggaac gttgcattaa atcgctttta 900
tttaacgttg atggccgcta cgtgctcgtt ctggtgcgcg gcgatcatga agcgaatgat 960
gtgaaagtga aaaatgtgct tgatgcgact gtcgtggagc tggcgacacc ggaagaaaca 1020
gcacgagtga tgaactgccc ggttggttcg ctcggcccga ttggcgtcag cgaagaggtg 1080
acgattatcg ccgatcatgc tgtcgcggcg atcgtaaacg gcgtctgcgg cgccaatgag 1140
gaaggatacc attatacggg tgtcaatcca gaccgcgatt ttgccgtcag tcaatatgcg 1200
gatttgcgtt tcgtccaaga aggcgaccct tctccggatg gcaacgggac gatccgcttc 1260
gctcgtggca ttgaagttgg acatgtgttt aagctcggta cgaaatatag cgaggcgatg 1320
aacgccgttt acctcgacga aaatggtcgg acacagacga tgattatggg ttgctacggc 1380
attggcgtct ctaggctcgt tgcggcgatc gccgagcagt tcgccgatga gaacgggctt 1440
gtatggccgg tttcggtcgc accgtttcac gttcatttgc tgacggcgaa cgcgaaaagc 1500
gatgaacagc gcatgctggc tgaagagtgg tacgaaaaac tcggacaggc cggatttgac 1560
gtgttgtatg atgaccgtcc ggaacgggcc ggggtgaagt ttgccgacag cgatttgatc 1620
ggcatcccgc tccgcgtcac cgttggcaag cgggcaagtg aaggtgtggt cgaagtaaaa 1680
gttcggaaaa caggcgagac gtttgacgtg ccggtcggtg agctgatcga aacagtgcgc 1740
cgtcttttgc aagga 1755
<210> 120
<211> 585
<212> PRT
<213> Geobacillus subterraneus
<400> 120
Met Thr Phe Lys Asn Ser Ser Tyr Asn Glu Arg Glu Lys Thr Arg Trp
1 5 10 15
Leu Leu Met Arg Gln Ser Gln Gly Phe Ile Pro Thr Leu Arg Glu Val
20 25 30
Pro Ala Asp Ala Glu Val Lys Ser His Gln Leu Leu Leu Arg Ala Gly
35 40 45
Phe Val Arg Gln Ser Ala Ser Gly Val Tyr Thr Phe Leu Pro Leu Gly
50 55 60
Gln Arg Val Leu Gln Lys Val Glu Ala Ile Ile Arg Glu Glu Met Asn
65 70 75 80
Arg Ala Gly Ala Leu Glu Leu Leu Met Pro Ala Leu Gln Pro Ala Glu
85 90 95
Leu Trp Gln Gln Ser Gly Arg Trp Tyr Ser Tyr Gly Pro Glu Leu Met
100 105 110
Arg Leu Lys Asp Arg His Glu Arg Asp Phe Val Leu Gly Pro Thr His
115 120 125
Glu Glu Met Ile Thr Thr Ile Val Arg Asp Glu Val Lys Thr Tyr Lys
130 135 140
Arg Leu Pro Leu Ile Leu Tyr Gln Ile Gln Thr Lys Phe Arg Asp Glu
145 150 155 160
Lys Arg Pro Arg Phe Gly Leu Leu Arg Gly Arg Glu Phe Ile Met Lys
165 170 175
Asp Ala Tyr Ser Phe His Thr Ser Gln Glu Ser Leu Asp Glu Thr Tyr
180 185 190
Asn Lys Met Tyr Glu Ala Tyr Ala Asn Ile Phe Arg Arg Cys Gly Leu
195 200 205
Asn Phe Arg Ala Val Ile Ala Asp Ser Gly Ala Met Gly Gly Lys Asp
210 215 220
Thr His Glu Phe Met Val Leu Ser Asp Ile Gly Glu Asp Thr Ile Ala
225 230 235 240
Tyr Ser Asp Ala Ser Asp Tyr Ala Ala Asn Ile Glu Met Ala Pro Val
245 250 255
Val Thr Thr Tyr Glu Lys Ser Ser Glu Pro Leu Val Glu Leu Lys Lys
260 265 270
Val Ala Thr Pro Glu Gln Lys Thr Ile Ala Glu Val Ala Ser Tyr Leu
275 280 285
Gln Val Ala Pro Glu Arg Cys Ile Lys Ser Leu Leu Phe Asn Val Asp
290 295 300
Gly Arg Tyr Val Leu Val Leu Val Arg Gly Asp His Glu Ala Asn Asp
305 310 315 320
Val Lys Val Lys Asn Val Leu Asp Ala Thr Val Val Glu Leu Ala Thr
325 330 335
Pro Glu Glu Thr Ala Arg Val Met Asn Cys Pro Val Gly Ser Leu Gly
340 345 350
Pro Ile Gly Val Ser Glu Glu Val Thr Ile Ile Ala Asp His Ala Val
355 360 365
Ala Ala Ile Val Asn Gly Val Cys Gly Ala Asn Glu Glu Gly Tyr His
370 375 380
Tyr Thr Gly Val Asn Pro Asp Arg Asp Phe Ala Val Ser Gln Tyr Ala
385 390 395 400
Asp Leu Arg Phe Val Gln Glu Gly Asp Pro Ser Pro Asp Gly Asn Gly
405 410 415
Thr Ile Arg Phe Ala Arg Gly Ile Glu Val Gly His Val Phe Lys Leu
420 425 430
Gly Thr Lys Tyr Ser Glu Ala Met Asn Ala Val Tyr Leu Asp Glu Asn
435 440 445
Gly Arg Thr Gln Thr Met Ile Met Gly Cys Tyr Gly Ile Gly Val Ser
450 455 460
Arg Leu Val Ala Ala Ile Ala Glu Gln Phe Ala Asp Glu Asn Gly Leu
465 470 475 480
Val Trp Pro Val Ser Val Ala Pro Phe His Val His Leu Leu Thr Ala
485 490 495
Asn Ala Lys Ser Asp Glu Gln Arg Met Leu Ala Glu Glu Trp Tyr Glu
500 505 510
Lys Leu Gly Gln Ala Gly Phe Asp Val Leu Tyr Asp Asp Arg Pro Glu
515 520 525
Arg Ala Gly Val Lys Phe Ala Asp Ser Asp Leu Ile Gly Ile Pro Leu
530 535 540
Arg Val Thr Val Gly Lys Arg Ala Ser Glu Gly Val Val Glu Val Lys
545 550 555 560
Val Arg Lys Thr Gly Glu Thr Phe Asp Val Pro Val Gly Glu Leu Ile
565 570 575
Glu Thr Val Arg Arg Leu Leu Gln Gly
580 585
<210> 121
<211> 1296
<212> DNA
<213> Geobacillus subterraneus
<400> 121
atggtggata aggaggtaaa gcgaatgctg gatgtgaaat tactacgcac ccaatttcaa 60
gaggtgaaag aaaaactgct gcagcgcggc gacgacttgg ccaacatcga ccggtttgag 120
cagcttgata aagagcgtcg tcgtttgatc gctcaggtgg aggagttaaa aagcaagcgc 180
aatgaggtgt cgcaacaaat tgctgtctta aagcgtgaaa aaaaggacgc cgagtcgttg 240
atcgtcgaaa tgcgcgaagt cggcgaccgc attaaacaaa tggacgagca aattcgccaa 300
cttgaagaag agctcgacag ccttctgtta tcgattccga atgtaccgca tgagtcagtg 360
ccagtcggtc agtcggaaga agataatgtc gaagtgcgaa gatgggggga accgcgttcg 420
ttctcgttcg aaccgaagcc acattgggac attgctgacc aactcggttt gctcgatttt 480
gagcgggctg ccaaagtggc aggaagtcgg tttgtgtttt acaaaggact aggggctcgt 540
cttgagcggg cattaatcaa ctttatgctc gacatccatc tcgatgaatt tggctatcaa 600
gaggtgttgc cgccatactt agtgaaccgg gcgagcatga tcggaacagg gcaattgcca 660
aaatttgcgg aagacgcgtt ccacttggac aatgaagact attttctcat tccaacagcg 720
gaagtgcctg tgacgaattt gcatcgcgat gaaattttaa cggctgatga cttgccgctt 780
tactatgcgg cttacagcgc gtgcttccgc gccgaagctg gctcggctgg ccgtgacacg 840
cgggggctca tccgccagca ccaattcaat aaagtggagc tcgtcaagtt cgtcaagccg 900
gaggattcat atgacgagtt ggaaaaattg acgcaccaag ccgaaacgat cctgcaacgg 960
ctcggacttc cgtatcgcgt cgtagccttg tgtacagggg atctgggatt ttcagcggcg 1020
aagacgtatg atattgaggt gtggctgcca agctatggaa cgtatcggga aatttcgtcg 1080
tgcagcaact ttgaggcgtt ccaggcgcgc cgagctaata tccgcttccg tcgcgagccg 1140
aaagcaaagc cagaatatgt gcatacgcta aacggttcgg ggctagccat cggccgcacg 1200
gttgctgcca ttttggaaaa ctaccaacaa gaagacggat cggtcgtcat cccggaagcg 1260
ctccgtccat atatggggaa tcgggatgtc attcgc 1296
<210> 122
<211> 432
<212> PRT
<213> Geobacillus subterraneus
<400> 122
Met Val Asp Lys Glu Val Lys Arg Met Leu Asp Val Lys Leu Leu Arg
1 5 10 15
Thr Gln Phe Gln Glu Val Lys Glu Lys Leu Leu Gln Arg Gly Asp Asp
20 25 30
Leu Ala Asn Ile Asp Arg Phe Glu Gln Leu Asp Lys Glu Arg Arg Arg
35 40 45
Leu Ile Ala Gln Val Glu Glu Leu Lys Ser Lys Arg Asn Glu Val Ser
50 55 60
Gln Gln Ile Ala Val Leu Lys Arg Glu Lys Lys Asp Ala Glu Ser Leu
65 70 75 80
Ile Val Glu Met Arg Glu Val Gly Asp Arg Ile Lys Gln Met Asp Glu
85 90 95
Gln Ile Arg Gln Leu Glu Glu Glu Leu Asp Ser Leu Leu Leu Ser Ile
100 105 110
Pro Asn Val Pro His Glu Ser Val Pro Val Gly Gln Ser Glu Glu Asp
115 120 125
Asn Val Glu Val Arg Arg Trp Gly Glu Pro Arg Ser Phe Ser Phe Glu
130 135 140
Pro Lys Pro His Trp Asp Ile Ala Asp Gln Leu Gly Leu Leu Asp Phe
145 150 155 160
Glu Arg Ala Ala Lys Val Ala Gly Ser Arg Phe Val Phe Tyr Lys Gly
165 170 175
Leu Gly Ala Arg Leu Glu Arg Ala Leu Ile Asn Phe Met Leu Asp Ile
180 185 190
His Leu Asp Glu Phe Gly Tyr Gln Glu Val Leu Pro Pro Tyr Leu Val
195 200 205
Asn Arg Ala Ser Met Ile Gly Thr Gly Gln Leu Pro Lys Phe Ala Glu
210 215 220
Asp Ala Phe His Leu Asp Asn Glu Asp Tyr Phe Leu Ile Pro Thr Ala
225 230 235 240
Glu Val Pro Val Thr Asn Leu His Arg Asp Glu Ile Leu Thr Ala Asp
245 250 255
Asp Leu Pro Leu Tyr Tyr Ala Ala Tyr Ser Ala Cys Phe Arg Ala Glu
260 265 270
Ala Gly Ser Ala Gly Arg Asp Thr Arg Gly Leu Ile Arg Gln His Gln
275 280 285
Phe Asn Lys Val Glu Leu Val Lys Phe Val Lys Pro Glu Asp Ser Tyr
290 295 300
Asp Glu Leu Glu Lys Leu Thr His Gln Ala Glu Thr Ile Leu Gln Arg
305 310 315 320
Leu Gly Leu Pro Tyr Arg Val Val Ala Leu Cys Thr Gly Asp Leu Gly
325 330 335
Phe Ser Ala Ala Lys Thr Tyr Asp Ile Glu Val Trp Leu Pro Ser Tyr
340 345 350
Gly Thr Tyr Arg Glu Ile Ser Ser Cys Ser Asn Phe Glu Ala Phe Gln
355 360 365
Ala Arg Arg Ala Asn Ile Arg Phe Arg Arg Glu Pro Lys Ala Lys Pro
370 375 380
Glu Tyr Val His Thr Leu Asn Gly Ser Gly Leu Ala Ile Gly Arg Thr
385 390 395 400
Val Ala Ala Ile Leu Glu Asn Tyr Gln Gln Glu Asp Gly Ser Val Val
405 410 415
Ile Pro Glu Ala Leu Arg Pro Tyr Met Gly Asn Arg Asp Val Ile Arg
420 425 430
<210> 123
<211> 1947
<212> DNA
<213> Geobacillus subterraneus
<400> 123
atgccagacg ttattcgcat tacgttcccg gacggggcga aaaaggagtt tccgagcgga 60
acgtcaactg aggacatcgc tgcctcgatc agtccgggat tgaagaaaaa agcgattgcc 120
gggaaactga acggccggtt tgttgattta cgcacgccgc ttcaagaaga cggcgagctt 180
gtcattatta cccaggacat gcctgaggca cttgatattt tgcgtcatag caccgcccat 240
ttaatggcgc aagcgatcaa gcggctgtat gacaacgtca agcttggcgt cggcccggtc 300
attgaaaacg gcttctacta tgatattgat atggaacata agctgacgcc ggatgatttg 360
ccgaaaattg aggcggaaat gcgcaaaatc gtaaaggaaa atcttgacgt tgttcgcaaa 420
gaggtgagcc gtgacgaggc gattcgcctg tatgaaaaaa ttggtgatca cttgaaactg 480
gagctcatca acgatattcc ggaaggcgag acgatttcca tttacgagca aggcgagttt 540
ttcgatcttt gtcggggtgt gcacgtgccg tcgaccggga aaatcaaaga gttcaagctg 600
ctcagcatct cgggggccta ctggcgcggt gacagcaaca acaaaatgct gcagcgtatt 660
tacggtacgg cgtttttcaa aaaagaagat ctggaccatt atttgcagtt gctcgaagag 720
gcgaaagagc gcgatcatcg caaattgggc aaagagcttg agctatttac gacatcacaa 780
aaagtcggac aaggactgcc gctttggttg ccgaaagggg cgacgatccg tcgcttgatt 840
gaacggtaca ttgtcgataa agaaatcgcc cttggttatg atcatgtata tacgccggtg 900
ctcggcagtg tggagctgta taaaacctca ggacactggg accattataa agaaaacatg 960
ttcccaccga tggaaatgga taacgaagag ctcgtgctgc ggccgatgaa ctgcccgcac 1020
catatgatga tttataaaag caagcttcat agctaccgtg agctgccgat ccgcatcgcc 1080
gagctcggca cgatgcatcg ctacgaaatg tccggggcgc ttactggact gcagcgtgtc 1140
cgcggcatga cgctcaacga cgcccatatt ttcgtgcgcc cggatcaaat taaagacgag 1200
tttaagcgcg tcgttaattt gattttggaa gtatacaaag actttgggct ggacgaatat 1260
tcgttccgcc tgtcgtaccg cgacccacaa gataaagaaa aatattacga cgacgacgag 1320
atgtgggaaa aggcgcaacg catgctgcgc gaggcgatgg atgaacttgg cctcgattac 1380
tacgaagcgg aaggggaagc agcgttttac ggaccgaagc tcgatgtgca agtgcgcacg 1440
gcactcggca aagatgagac gctgtcgact gtacagcttg acttcctctt gccggagcgg 1500
tttgacttaa catatatcgg cgaagatgga aaaccgcacc gcccggtcgt catccaccgc 1560
ggcgttgttt ccacgatgga acggtttgtc gccttcttga tcgaagaata caaaggggca 1620
tttccaacgt ggctcgcccc ggtgcaagtg gaagtcatcc cggtatcgtc ggaagcccat 1680
ctcgattatg cgtatgaagt gaaacaagcg ctgcaagtaa acggcttccg cgtcgaagtc 1740
gacgaacggg atgaaaaaat cggctataaa atccgcgaag cgcaaatgca aaaaattcct 1800
tatatgctcg ttgtcggcga caaagaagcg gccgagcgag cggtcaacgt ccgccgctac 1860
ggtgaaaaag aaagcgagac tgtggcgctt gacaagttta tcgcgatgct agaagaagat 1920
gtgcggcaaa aacgagtgaa aaaacga 1947
<210> 124
<211> 649
<212> PRT
<213> Geobacillus subterraneus
<400> 124
Met Pro Asp Val Ile Arg Ile Thr Phe Pro Asp Gly Ala Lys Lys Glu
1 5 10 15
Phe Pro Ser Gly Thr Ser Thr Glu Asp Ile Ala Ala Ser Ile Ser Pro
20 25 30
Gly Leu Lys Lys Lys Ala Ile Ala Gly Lys Leu Asn Gly Arg Phe Val
35 40 45
Asp Leu Arg Thr Pro Leu Gln Glu Asp Gly Glu Leu Val Ile Ile Thr
50 55 60
Gln Asp Met Pro Glu Ala Leu Asp Ile Leu Arg His Ser Thr Ala His
65 70 75 80
Leu Met Ala Gln Ala Ile Lys Arg Leu Tyr Asp Asn Val Lys Leu Gly
85 90 95
Val Gly Pro Val Ile Glu Asn Gly Phe Tyr Tyr Asp Ile Asp Met Glu
100 105 110
His Lys Leu Thr Pro Asp Asp Leu Pro Lys Ile Glu Ala Glu Met Arg
115 120 125
Lys Ile Val Lys Glu Asn Leu Asp Val Val Arg Lys Glu Val Ser Arg
130 135 140
Asp Glu Ala Ile Arg Leu Tyr Glu Lys Ile Gly Asp His Leu Lys Leu
145 150 155 160
Glu Leu Ile Asn Asp Ile Pro Glu Gly Glu Thr Ile Ser Ile Tyr Glu
165 170 175
Gln Gly Glu Phe Phe Asp Leu Cys Arg Gly Val His Val Pro Ser Thr
180 185 190
Gly Lys Ile Lys Glu Phe Lys Leu Leu Ser Ile Ser Gly Ala Tyr Trp
195 200 205
Arg Gly Asp Ser Asn Asn Lys Met Leu Gln Arg Ile Tyr Gly Thr Ala
210 215 220
Phe Phe Lys Lys Glu Asp Leu Asp His Tyr Leu Gln Leu Leu Glu Glu
225 230 235 240
Ala Lys Glu Arg Asp His Arg Lys Leu Gly Lys Glu Leu Glu Leu Phe
245 250 255
Thr Thr Ser Gln Lys Val Gly Gln Gly Leu Pro Leu Trp Leu Pro Lys
260 265 270
Gly Ala Thr Ile Arg Arg Leu Ile Glu Arg Tyr Ile Val Asp Lys Glu
275 280 285
Ile Ala Leu Gly Tyr Asp His Val Tyr Thr Pro Val Leu Gly Ser Val
290 295 300
Glu Leu Tyr Lys Thr Ser Gly His Trp Asp His Tyr Lys Glu Asn Met
305 310 315 320
Phe Pro Pro Met Glu Met Asp Asn Glu Glu Leu Val Leu Arg Pro Met
325 330 335
Asn Cys Pro His His Met Met Ile Tyr Lys Ser Lys Leu His Ser Tyr
340 345 350
Arg Glu Leu Pro Ile Arg Ile Ala Glu Leu Gly Thr Met His Arg Tyr
355 360 365
Glu Met Ser Gly Ala Leu Thr Gly Leu Gln Arg Val Arg Gly Met Thr
370 375 380
Leu Asn Asp Ala His Ile Phe Val Arg Pro Asp Gln Ile Lys Asp Glu
385 390 395 400
Phe Lys Arg Val Val Asn Leu Ile Leu Glu Val Tyr Lys Asp Phe Gly
405 410 415
Leu Asp Glu Tyr Ser Phe Arg Leu Ser Tyr Arg Asp Pro Gln Asp Lys
420 425 430
Glu Lys Tyr Tyr Asp Asp Asp Glu Met Trp Glu Lys Ala Gln Arg Met
435 440 445
Leu Arg Glu Ala Met Asp Glu Leu Gly Leu Asp Tyr Tyr Glu Ala Glu
450 455 460
Gly Glu Ala Ala Phe Tyr Gly Pro Lys Leu Asp Val Gln Val Arg Thr
465 470 475 480
Ala Leu Gly Lys Asp Glu Thr Leu Ser Thr Val Gln Leu Asp Phe Leu
485 490 495
Leu Pro Glu Arg Phe Asp Leu Thr Tyr Ile Gly Glu Asp Gly Lys Pro
500 505 510
His Arg Pro Val Val Ile His Arg Gly Val Val Ser Thr Met Glu Arg
515 520 525
Phe Val Ala Phe Leu Ile Glu Glu Tyr Lys Gly Ala Phe Pro Thr Trp
530 535 540
Leu Ala Pro Val Gln Val Glu Val Ile Pro Val Ser Ser Glu Ala His
545 550 555 560
Leu Asp Tyr Ala Tyr Glu Val Lys Gln Ala Leu Gln Val Asn Gly Phe
565 570 575
Arg Val Glu Val Asp Glu Arg Asp Glu Lys Ile Gly Tyr Lys Ile Arg
580 585 590
Glu Ala Gln Met Gln Lys Ile Pro Tyr Met Leu Val Val Gly Asp Lys
595 600 605
Glu Ala Ala Glu Arg Ala Val Asn Val Arg Arg Tyr Gly Glu Lys Glu
610 615 620
Ser Glu Thr Val Ala Leu Asp Lys Phe Ile Ala Met Leu Glu Glu Asp
625 630 635 640
Val Arg Gln Lys Arg Val Lys Lys Arg
645
<210> 125
<211> 984
<212> DNA
<213> Geobacillus subterraneus
<400> 125
atgaaaacca ttttttctgg cattcagcca agcggcgtca ttacccttgg caactacatt 60
ggtgcgatgc gacaatttgt cgaactgcag catgagtaca actgctattt ttgcattgtc 120
gaccaacatg ccattactgt tccgcaaaat ccgaacgaac tgcaacaaaa cattcgccgt 180
ctcgctgcct tatatttggc agtcggcatc gatcctaaac aggcgacgct gttcgttcaa 240
tcggaggtgc cggcgcacgc ccaagcggct tggatgctgc aatgcatcgt ctatatcggc 300
gaactggagc ggatgacgca gtttaaagac aaatcagccg gtaaagaggc ggtcagtgcc 360
gggttgctca cgtatccacc gcttatggca gccgacattt tgctttacaa cacggacatt 420
gtcccagtcg gcgaagacca aaagcagcac atcgagctga cgcgcgattt agctgagcgc 480
ttcaacaaac ggtacggcga gctgttcact atcccggaag cgcgcatccc gaaaatcggc 540
gcccgcatta tgtcgcttac cgatccgacg aaaaaaatga gcaaatctga cccaaacccg 600
aaatcgttta ttacgctgct tgacgacgcc aaaacgattg aaaagaaaat taaaagtgct 660
gtgaccgatt cagaaggaac gattcgctat gacaaggaag cgaaaccggg catttcgaac 720
ttgctcaaca tttattcgat tttatcgggt cagccgattg acgaacttga gcggcaatac 780
gaaggaaaag gatacggggt ctttaaatcc gatttggccc aagtggtcat tgaaacgctc 840
caaccgatcc aagagcggta ttatcattgg ctcgaaagtg aagagctcga ccgcgtccta 900
gacgaagggg cggaaaaagc gaaccgtgtc gcctcggaaa tggtgcgcaa aatggaacaa 960
gccatggggc ttgggcggcg tcgg 984
<210> 126
<211> 328
<212> PRT
<213> Geobacillus subterraneus
<400> 126
Met Lys Thr Ile Phe Ser Gly Ile Gln Pro Ser Gly Val Ile Thr Leu
1 5 10 15
Gly Asn Tyr Ile Gly Ala Met Arg Gln Phe Val Glu Leu Gln His Glu
20 25 30
Tyr Asn Cys Tyr Phe Cys Ile Val Asp Gln His Ala Ile Thr Val Pro
35 40 45
Gln Asn Pro Asn Glu Leu Gln Gln Asn Ile Arg Arg Leu Ala Ala Leu
50 55 60
Tyr Leu Ala Val Gly Ile Asp Pro Lys Gln Ala Thr Leu Phe Val Gln
65 70 75 80
Ser Glu Val Pro Ala His Ala Gln Ala Ala Trp Met Leu Gln Cys Ile
85 90 95
Val Tyr Ile Gly Glu Leu Glu Arg Met Thr Gln Phe Lys Asp Lys Ser
100 105 110
Ala Gly Lys Glu Ala Val Ser Ala Gly Leu Leu Thr Tyr Pro Pro Leu
115 120 125
Met Ala Ala Asp Ile Leu Leu Tyr Asn Thr Asp Ile Val Pro Val Gly
130 135 140
Glu Asp Gln Lys Gln His Ile Glu Leu Thr Arg Asp Leu Ala Glu Arg
145 150 155 160
Phe Asn Lys Arg Tyr Gly Glu Leu Phe Thr Ile Pro Glu Ala Arg Ile
165 170 175
Pro Lys Ile Gly Ala Arg Ile Met Ser Leu Thr Asp Pro Thr Lys Lys
180 185 190
Met Ser Lys Ser Asp Pro Asn Pro Lys Ser Phe Ile Thr Leu Leu Asp
195 200 205
Asp Ala Lys Thr Ile Glu Lys Lys Ile Lys Ser Ala Val Thr Asp Ser
210 215 220
Glu Gly Thr Ile Arg Tyr Asp Lys Glu Ala Lys Pro Gly Ile Ser Asn
225 230 235 240
Leu Leu Asn Ile Tyr Ser Ile Leu Ser Gly Gln Pro Ile Asp Glu Leu
245 250 255
Glu Arg Gln Tyr Glu Gly Lys Gly Tyr Gly Val Phe Lys Ser Asp Leu
260 265 270
Ala Gln Val Val Ile Glu Thr Leu Gln Pro Ile Gln Glu Arg Tyr Tyr
275 280 285
His Trp Leu Glu Ser Glu Glu Leu Asp Arg Val Leu Asp Glu Gly Ala
290 295 300
Glu Lys Ala Asn Arg Val Ala Ser Glu Met Val Arg Lys Met Glu Gln
305 310 315 320
Ala Met Gly Leu Gly Arg Arg Arg
325
<210> 127
<211> 1257
<212> DNA
<213> Geobacillus subterraneus
<400> 127
atgaacctgc ttgaagaact gcaatggcgc ggacttgtca atcaaacgac ggatgaggat 60
gggcttcgaa agctcctgaa tgaggagaag gtgacgcttt attgcgggtt tgacccgaca 120
gcagacagct tgcatatcgg ccatttggtc acgatcatga ccttgcgtcg tttccaacag 180
gcggggcatc aaccgatcgc cttagtcggc ggcgccaccg ggttgatcgg cgatccgagt 240
ggcagaaaaa gcgagcgcac gctcaacgcc aaggagacgg tcgagacgtg gagcgcccga 300
atcaaagcgc aactcgagcg gtttcttgat tttgaggctg agagcaatcc agcgaaaatc 360
aaaaacaact acgactggat cgggccgctt gatgtcatct cgtttttgcg tgacatcggc 420
aagcatttca gcgtcaatta catgcttgcg aaagaatcgg tgcagtcgcg cattgaaatg 480
ggcatttcgt ttaccgagtt cagctatatg atgctgcagg cgtacgactt cctcaacttg 540
tacgaaacgg aaggttgccg actacaaatc ggtggcagcg accaatgggg caacatcacg 600
gcggggcttg agctcatccg cagaacgaaa ggtgaggcga aagcatttgg tttgacggtt 660
ccgctcgtga cgaaagccga tgggacgaag ttcggaaaaa cggaaagcgg cgcggtttgg 720
ctcgatccgg aaaaaacgtc gccgtatgag ttttaccagt tctggatcaa caccgatgac 780
cgcgatgtga tccgttactt aaaatatttc acgttcttga caaaagaaga gatcgacgcg 840
cttgaacaag agctgcgcga agcgccggag aagcgggtgg cgcaaaaaac gcttgcttcc 900
gaagtgacga agctcgtgca tggcgaagag gcgctcaatc aagcgattcg tatttcagaa 960
gcactcttta gcggcgacat tgccgaactg acggctgcgg aaatcgagca agggtttaaa 1020
aacgtgccgt cgtttgtcca tgaaggaggc gacgtcccgc tcgtcgagct gctcgtagct 1080
gccggcatct cgccatcgaa gcggcaggcg cgcgaagatg ttcaaaacgg tgcgatttat 1140
gtcaacggcg agcgcatcca agatgtcggc gctgtcttaa cggccgaaca ccgtttggaa 1200
gggcggttta ccgtgatccg ccgcggcaag aagaagtatt atttaatccg ctacgct 1257
<210> 128
<211> 419
<212> PRT
<213> Geobacillus subterraneus
<400> 128
Met Asn Leu Leu Glu Glu Leu Gln Trp Arg Gly Leu Val Asn Gln Thr
1 5 10 15
Thr Asp Glu Asp Gly Leu Arg Lys Leu Leu Asn Glu Glu Lys Val Thr
20 25 30
Leu Tyr Cys Gly Phe Asp Pro Thr Ala Asp Ser Leu His Ile Gly His
35 40 45
Leu Val Thr Ile Met Thr Leu Arg Arg Phe Gln Gln Ala Gly His Gln
50 55 60
Pro Ile Ala Leu Val Gly Gly Ala Thr Gly Leu Ile Gly Asp Pro Ser
65 70 75 80
Gly Arg Lys Ser Glu Arg Thr Leu Asn Ala Lys Glu Thr Val Glu Thr
85 90 95
Trp Ser Ala Arg Ile Lys Ala Gln Leu Glu Arg Phe Leu Asp Phe Glu
100 105 110
Ala Glu Ser Asn Pro Ala Lys Ile Lys Asn Asn Tyr Asp Trp Ile Gly
115 120 125
Pro Leu Asp Val Ile Ser Phe Leu Arg Asp Ile Gly Lys His Phe Ser
130 135 140
Val Asn Tyr Met Leu Ala Lys Glu Ser Val Gln Ser Arg Ile Glu Met
145 150 155 160
Gly Ile Ser Phe Thr Glu Phe Ser Tyr Met Met Leu Gln Ala Tyr Asp
165 170 175
Phe Leu Asn Leu Tyr Glu Thr Glu Gly Cys Arg Leu Gln Ile Gly Gly
180 185 190
Ser Asp Gln Trp Gly Asn Ile Thr Ala Gly Leu Glu Leu Ile Arg Arg
195 200 205
Thr Lys Gly Glu Ala Lys Ala Phe Gly Leu Thr Val Pro Leu Val Thr
210 215 220
Lys Ala Asp Gly Thr Lys Phe Gly Lys Thr Glu Ser Gly Ala Val Trp
225 230 235 240
Leu Asp Pro Glu Lys Thr Ser Pro Tyr Glu Phe Tyr Gln Phe Trp Ile
245 250 255
Asn Thr Asp Asp Arg Asp Val Ile Arg Tyr Leu Lys Tyr Phe Thr Phe
260 265 270
Leu Thr Lys Glu Glu Ile Asp Ala Leu Glu Gln Glu Leu Arg Glu Ala
275 280 285
Pro Glu Lys Arg Val Ala Gln Lys Thr Leu Ala Ser Glu Val Thr Lys
290 295 300
Leu Val His Gly Glu Glu Ala Leu Asn Gln Ala Ile Arg Ile Ser Glu
305 310 315 320
Ala Leu Phe Ser Gly Asp Ile Ala Glu Leu Thr Ala Ala Glu Ile Glu
325 330 335
Gln Gly Phe Lys Asn Val Pro Ser Phe Val His Glu Gly Gly Asp Val
340 345 350
Pro Leu Val Glu Leu Leu Val Ala Ala Gly Ile Ser Pro Ser Lys Arg
355 360 365
Gln Ala Arg Glu Asp Val Gln Asn Gly Ala Ile Tyr Val Asn Gly Glu
370 375 380
Arg Ile Gln Asp Val Gly Ala Val Leu Thr Ala Glu His Arg Leu Glu
385 390 395 400
Gly Arg Phe Thr Val Ile Arg Arg Gly Lys Lys Lys Tyr Tyr Leu Ile
405 410 415
Arg Tyr Ala
<210> 129
<211> 2727
<212> DNA
<213> Geobacillus subterraneus
<400> 129
atgaaagggg cttttttgct tgcctatcgg acggttgatc ctgtaggcaa cacagccatt 60
gtttatcaca tgaaggaggg aataaaagtg gcacagcatg aagtgtcgat gccgccaaaa 120
tacgatcacc gcgctgttga agcggggcgc tatgactggt ggctgaaagg caagtttttt 180
gaaacgaccg gcgatccgga caaacaaccg tttacgatcg ttatcccacc gccgaacgtc 240
acaggcaaac tgcatttggg ccatgcgtgg gatacgacgc tgcaagacat cattacgcgc 300
atgaagcgga tgcaagggta tgatgtccta tggcttccgg gtatggacca tgccggcatc 360
gccacccagg cgaaagtgga agaaaaattg cgccaacaag gactgtcccg ctacgattta 420
ggacgggaaa aatttttgga agaaacgtgg aaatggaaag aagaatatgc cggccatatc 480
cgcagccaat gggcaaaatt agggctcggc ctcgattaca cgcgcgagcg gtttacgctt 540
gatgaagggc tgtcaaaagc cgtacgcgaa gtgttcgtct cgctttaccg gaaagggctc 600
atttaccgcg gtgaatacat tatcaactgg gatccggcga ccaaaaccgc cttgtccgac 660
atcgaggtca tttacaagga agtgaaaggt gcgctttatc atttgcgcta tccgctcgct 720
gacggctcgg gctacattga agtagcgaca acccgtccag aaacgatgct cggtgacacg 780
gccgtcgcgg ttcatccgga tgacgagcgg tataaacact tgatcggcaa gatggtgaaa 840
ttgccaatcg ttggccggga aattccgatc atcgctgatg agtatgtcga tatggaattc 900
ggttccggcg cggtaaaaat tacaccggca cacgatccga acgactttga agttggcaac 960
cgccacaact tgccgcgcat tctcgtcatg aacgaagacg gtacaatgaa cgaaaacgca 1020
ttgcaatatc aagggcttga ccggtttgaa tgccggaagc aaatcgtccg tgatttacaa 1080
gagcaaggcg tcctctttaa aattgaggaa cacgtccact cggtcgggca cagtgaacgg 1140
agcggcgccg ttgttgaacc gtatttgtcg acacaatggt tcgtaaaaat gaagccgctc 1200
gcggaagctg ccatcaagat gcagcaaaca gaaggaaaag tgcaatttgt gccggagcgg 1260
tttgaaaaaa cgtacttgca ctggcttgag aacattcgcg actggtgcat ttcgcgtcag 1320
ctttggtggg ggcaccgcat tccggcgtgg taccataaag aaacgggtga aatttacgtc 1380
gaccacgagc cgccggcaga cattgaaaat tgggagcaag acccggatgt gcttgataca 1440
tggttcagct cggcactctg gccgttctcc acaatggggt ggccggatac ggaagcgccg 1500
gactacaagc gctattaccc gaccgatgtg cttgtcaccg gctatgacat cattttcttc 1560
tgggtgtcgc gcatgatttt ccaagggctt gagttcactg ggaagagacc gtttaaagat 1620
gtgttgatcc acggcctcgt ccgcgacgct caaggaagaa aaatgagcaa gtcgctcggc 1680
aacggtgtcg acccgatgga tgtcattgac caatacggcg ccgatgcgct ccgctacttc 1740
ctagcgaccg gtagctcgcc aggacaagat ttgcgcttta gcacggaaaa agttgaggcg 1800
acgtggaatt ttgctaacaa aatttggaac gcttcacgtt tcgccttaat gaacatgggc 1860
ggcatgacat atgaggagct cgatttgagc ggcgaaaaaa cggtcgccga ccattggatt 1920
ttaacgcgct taaatgaaac gatcgacacg gtgacgaagc tcgccgacaa atacgagttt 1980
ggtgaagtcg gtcgcacgtt gtacaacttt atttgggacg atttgtgcga ctggtacatt 2040
gaaatggcga agctgccgct ttacggcgat gatgagacag cgaaaaagac gacgcgttca 2100
gttttagcgt atgtgcttga caatacgatg cgcttgttgc atccattcat gccgttcatt 2160
accgaggaaa tttggcaaaa cttgccgcat gacggcgaat cgattaccgt tgcctcgtgg 2220
ccgcaagtgc gtccggagct gtcaaacgaa gaagcggcgg aagaaatgcg gatgctcgtt 2280
gacattatcc gcgcggtccg aaacgttcgt gccgaagtca atacgccgcc gagcaaaccg 2340
attgcgctct acattaagac aaaagacgaa caagtgcgcg cagcgcttat gaaaaaccgc 2400
gcttatctcg aacggttctg caatccgagc gaattgatca ttgacacgga tgttccggcg 2460
ccagaaaaag cgatgactgc tgtcgtcaca ggggcagagc tcattttgcc gcttgaagga 2520
ctcatcaata tcgaagaaga aatcaagcgg cttgagaaag agctcgacaa atggaacaaa 2580
gaagtcgagc gtgtcgaaaa gaaactggcg aacgaaggct ttttggcaaa agcgccggct 2640
catgtcgtcg aggaagagcg gcgcaagcgg caagattaca tcgaaaaacg cgaagcagtg 2700
aaagcgcgtc ttgccgagtt gaaacgg 2727
<210> 130
<211> 909
<212> PRT
<213> Geobacillus subterraneus
<400> 130
Met Lys Gly Ala Phe Leu Leu Ala Tyr Arg Thr Val Asp Pro Val Gly
1 5 10 15
Asn Thr Ala Ile Val Tyr His Met Lys Glu Gly Ile Lys Val Ala Gln
20 25 30
His Glu Val Ser Met Pro Pro Lys Tyr Asp His Arg Ala Val Glu Ala
35 40 45
Gly Arg Tyr Asp Trp Trp Leu Lys Gly Lys Phe Phe Glu Thr Thr Gly
50 55 60
Asp Pro Asp Lys Gln Pro Phe Thr Ile Val Ile Pro Pro Pro Asn Val
65 70 75 80
Thr Gly Lys Leu His Leu Gly His Ala Trp Asp Thr Thr Leu Gln Asp
85 90 95
Ile Ile Thr Arg Met Lys Arg Met Gln Gly Tyr Asp Val Leu Trp Leu
100 105 110
Pro Gly Met Asp His Ala Gly Ile Ala Thr Gln Ala Lys Val Glu Glu
115 120 125
Lys Leu Arg Gln Gln Gly Leu Ser Arg Tyr Asp Leu Gly Arg Glu Lys
130 135 140
Phe Leu Glu Glu Thr Trp Lys Trp Lys Glu Glu Tyr Ala Gly His Ile
145 150 155 160
Arg Ser Gln Trp Ala Lys Leu Gly Leu Gly Leu Asp Tyr Thr Arg Glu
165 170 175
Arg Phe Thr Leu Asp Glu Gly Leu Ser Lys Ala Val Arg Glu Val Phe
180 185 190
Val Ser Leu Tyr Arg Lys Gly Leu Ile Tyr Arg Gly Glu Tyr Ile Ile
195 200 205
Asn Trp Asp Pro Ala Thr Lys Thr Ala Leu Ser Asp Ile Glu Val Ile
210 215 220
Tyr Lys Glu Val Lys Gly Ala Leu Tyr His Leu Arg Tyr Pro Leu Ala
225 230 235 240
Asp Gly Ser Gly Tyr Ile Glu Val Ala Thr Thr Arg Pro Glu Thr Met
245 250 255
Leu Gly Asp Thr Ala Val Ala Val His Pro Asp Asp Glu Arg Tyr Lys
260 265 270
His Leu Ile Gly Lys Met Val Lys Leu Pro Ile Val Gly Arg Glu Ile
275 280 285
Pro Ile Ile Ala Asp Glu Tyr Val Asp Met Glu Phe Gly Ser Gly Ala
290 295 300
Val Lys Ile Thr Pro Ala His Asp Pro Asn Asp Phe Glu Val Gly Asn
305 310 315 320
Arg His Asn Leu Pro Arg Ile Leu Val Met Asn Glu Asp Gly Thr Met
325 330 335
Asn Glu Asn Ala Leu Gln Tyr Gln Gly Leu Asp Arg Phe Glu Cys Arg
340 345 350
Lys Gln Ile Val Arg Asp Leu Gln Glu Gln Gly Val Leu Phe Lys Ile
355 360 365
Glu Glu His Val His Ser Val Gly His Ser Glu Arg Ser Gly Ala Val
370 375 380
Val Glu Pro Tyr Leu Ser Thr Gln Trp Phe Val Lys Met Lys Pro Leu
385 390 395 400
Ala Glu Ala Ala Ile Lys Met Gln Gln Thr Glu Gly Lys Val Gln Phe
405 410 415
Val Pro Glu Arg Phe Glu Lys Thr Tyr Leu His Trp Leu Glu Asn Ile
420 425 430
Arg Asp Trp Cys Ile Ser Arg Gln Leu Trp Trp Gly His Arg Ile Pro
435 440 445
Ala Trp Tyr His Lys Glu Thr Gly Glu Ile Tyr Val Asp His Glu Pro
450 455 460
Pro Ala Asp Ile Glu Asn Trp Glu Gln Asp Pro Asp Val Leu Asp Thr
465 470 475 480
Trp Phe Ser Ser Ala Leu Trp Pro Phe Ser Thr Met Gly Trp Pro Asp
485 490 495
Thr Glu Ala Pro Asp Tyr Lys Arg Tyr Tyr Pro Thr Asp Val Leu Val
500 505 510
Thr Gly Tyr Asp Ile Ile Phe Phe Trp Val Ser Arg Met Ile Phe Gln
515 520 525
Gly Leu Glu Phe Thr Gly Lys Arg Pro Phe Lys Asp Val Leu Ile His
530 535 540
Gly Leu Val Arg Asp Ala Gln Gly Arg Lys Met Ser Lys Ser Leu Gly
545 550 555 560
Asn Gly Val Asp Pro Met Asp Val Ile Asp Gln Tyr Gly Ala Asp Ala
565 570 575
Leu Arg Tyr Phe Leu Ala Thr Gly Ser Ser Pro Gly Gln Asp Leu Arg
580 585 590
Phe Ser Thr Glu Lys Val Glu Ala Thr Trp Asn Phe Ala Asn Lys Ile
595 600 605
Trp Asn Ala Ser Arg Phe Ala Leu Met Asn Met Gly Gly Met Thr Tyr
610 615 620
Glu Glu Leu Asp Leu Ser Gly Glu Lys Thr Val Ala Asp His Trp Ile
625 630 635 640
Leu Thr Arg Leu Asn Glu Thr Ile Asp Thr Val Thr Lys Leu Ala Asp
645 650 655
Lys Tyr Glu Phe Gly Glu Val Gly Arg Thr Leu Tyr Asn Phe Ile Trp
660 665 670
Asp Asp Leu Cys Asp Trp Tyr Ile Glu Met Ala Lys Leu Pro Leu Tyr
675 680 685
Gly Asp Asp Glu Thr Ala Lys Lys Thr Thr Arg Ser Val Leu Ala Tyr
690 695 700
Val Leu Asp Asn Thr Met Arg Leu Leu His Pro Phe Met Pro Phe Ile
705 710 715 720
Thr Glu Glu Ile Trp Gln Asn Leu Pro His Asp Gly Glu Ser Ile Thr
725 730 735
Val Ala Ser Trp Pro Gln Val Arg Pro Glu Leu Ser Asn Glu Glu Ala
740 745 750
Ala Glu Glu Met Arg Met Leu Val Asp Ile Ile Arg Ala Val Arg Asn
755 760 765
Val Arg Ala Glu Val Asn Thr Pro Pro Ser Lys Pro Ile Ala Leu Tyr
770 775 780
Ile Lys Thr Lys Asp Glu Gln Val Arg Ala Ala Leu Met Lys Asn Arg
785 790 795 800
Ala Tyr Leu Glu Arg Phe Cys Asn Pro Ser Glu Leu Ile Ile Asp Thr
805 810 815
Asp Val Pro Ala Pro Glu Lys Ala Met Thr Ala Val Val Thr Gly Ala
820 825 830
Glu Leu Ile Leu Pro Leu Glu Gly Leu Ile Asn Ile Glu Glu Glu Ile
835 840 845
Lys Arg Leu Glu Lys Glu Leu Asp Lys Trp Asn Lys Glu Val Glu Arg
850 855 860
Val Glu Lys Lys Leu Ala Asn Glu Gly Phe Leu Ala Lys Ala Pro Ala
865 870 875 880
His Val Val Glu Glu Glu Arg Arg Lys Arg Gln Asp Tyr Ile Glu Lys
885 890 895
Arg Glu Ala Val Lys Ala Arg Leu Ala Glu Leu Lys Arg
900 905
<210> 131
<211> 963
<212> DNA
<213> Geobacillus subterraneus
<400> 131
atgctgatga cgaacattgt ctttatggga acgcctgatt ttgcggtgcc ggttttacgg 60
cagctgcttg atgacgggta tcgggttgtt gccgttgtta cgcagccgga caagccgaaa 120
gggcgaaagc gcgagcttgt tccgcccccc gttaaggtcg aggcgcaaaa acacggcatc 180
ccggtattgc aaccgacgaa aattcgtgaa ccggaacaat acgaacaagt gctggcgttt 240
gcgcctgact tgatcgtgac cgcggcattt ggacaaattt tgcctaaggc tctgcttgac 300
gctcccaaat atggctgcat taatgttcac gcctcgcttc ttcccgagct gcgcggcggt 360
gcgccgatcc attatgccat ttggcaaggg aaaacgaaaa caggtgtcac gattatgtat 420
atggcggaaa agttggatgc cggcgacatg ttgacgcaag tcgaagtgcc gattgaagaa 480
accgataccg tcggcacact gcatgataaa ttgagcgctg ccggggctaa actattatca 540
gaaacgctcc cgcttttatt ggaaggtaac cttgcgccta ttccgcaaga ggaagagaaa 600
gcgacatatg ctccgaatat ccggcgtgaa caagagcgga ttgactgggc gcagcctggt 660
gaggcgattt acaaccatat ccgtgctttt catccgtggc cggttacgta tacgacatac 720
gacgggaacg tttggaaaat ctggtggggc gaaaaagtgc cggcgccaag cttagcgtcg 780
ccaggcacga ttttatcgct tgaggaagac ggcatcgtcg tcgccaccgg cagtgagacg 840
gccattaaaa ttactgaatt gcagccggcc ggcaaaaagc gaatggcggc cagcgagttt 900
ttgcgcggtg ctggcagccg gcttgcggtc ggcacgaagc taggagagaa caatgaacgt 960
acg 963
<210> 132
<211> 321
<212> PRT
<213> Geobacillus subterraneus
<400> 132
Met Leu Met Thr Asn Ile Val Phe Met Gly Thr Pro Asp Phe Ala Val
1 5 10 15
Pro Val Leu Arg Gln Leu Leu Asp Asp Gly Tyr Arg Val Val Ala Val
20 25 30
Val Thr Gln Pro Asp Lys Pro Lys Gly Arg Lys Arg Glu Leu Val Pro
35 40 45
Pro Pro Val Lys Val Glu Ala Gln Lys His Gly Ile Pro Val Leu Gln
50 55 60
Pro Thr Lys Ile Arg Glu Pro Glu Gln Tyr Glu Gln Val Leu Ala Phe
65 70 75 80
Ala Pro Asp Leu Ile Val Thr Ala Ala Phe Gly Gln Ile Leu Pro Lys
85 90 95
Ala Leu Leu Asp Ala Pro Lys Tyr Gly Cys Ile Asn Val His Ala Ser
100 105 110
Leu Leu Pro Glu Leu Arg Gly Gly Ala Pro Ile His Tyr Ala Ile Trp
115 120 125
Gln Gly Lys Thr Lys Thr Gly Val Thr Ile Met Tyr Met Ala Glu Lys
130 135 140
Leu Asp Ala Gly Asp Met Leu Thr Gln Val Glu Val Pro Ile Glu Glu
145 150 155 160
Thr Asp Thr Val Gly Thr Leu His Asp Lys Leu Ser Ala Ala Gly Ala
165 170 175
Lys Leu Leu Ser Glu Thr Leu Pro Leu Leu Leu Glu Gly Asn Leu Ala
180 185 190
Pro Ile Pro Gln Glu Glu Glu Lys Ala Thr Tyr Ala Pro Asn Ile Arg
195 200 205
Arg Glu Gln Glu Arg Ile Asp Trp Ala Gln Pro Gly Glu Ala Ile Tyr
210 215 220
Asn His Ile Arg Ala Phe His Pro Trp Pro Val Thr Tyr Thr Thr Tyr
225 230 235 240
Asp Gly Asn Val Trp Lys Ile Trp Trp Gly Glu Lys Val Pro Ala Pro
245 250 255
Ser Leu Ala Ser Pro Gly Thr Ile Leu Ser Leu Glu Glu Asp Gly Ile
260 265 270
Val Val Ala Thr Gly Ser Glu Thr Ala Ile Lys Ile Thr Glu Leu Gln
275 280 285
Pro Ala Gly Lys Lys Arg Met Ala Ala Ser Glu Phe Leu Arg Gly Ala
290 295 300
Gly Ser Arg Leu Ala Val Gly Thr Lys Leu Gly Glu Asn Asn Glu Arg
305 310 315 320
Thr
<210> 133
<211> 358
<212> PRT
<213> Geobacillus stearothermophilus
<400> 133
Met Phe Asp Arg Leu Glu Ala Val Glu Gln Arg Tyr Glu Lys Leu Asn
1 5 10 15
Glu Leu Leu Met Glu Pro Asp Val Ile Asn Asp Pro Lys Lys Leu Arg
20 25 30
Asp Tyr Ser Lys Glu Gln Ala Asp Leu Gly Glu Thr Val Gln Thr Tyr
35 40 45
Arg Glu Tyr Lys Ser Val Arg Glu Gln Leu Ala Glu Ala Lys Ala Met
50 55 60
Leu Glu Glu Lys Leu Glu Pro Glu Leu Arg Glu Met Val Lys Glu Glu
65 70 75 80
Ile Gly Glu Leu Glu Glu Arg Glu Glu Ala Leu Val Glu Lys Leu Lys
85 90 95
Val Leu Leu Leu Pro Lys Asp Pro Asn Asp Glu Lys Asn Val Ile Met
100 105 110
Glu Ile Arg Ala Ala Ala Gly Gly Glu Glu Ala Ala Leu Phe Ala Gly
115 120 125
Asp Leu Tyr Arg Met Tyr Thr Arg Tyr Ala Glu Ser Gln Gly Trp Lys
130 135 140
Thr Glu Val Ile Glu Ala Ser Pro Thr Gly Leu Gly Gly Tyr Lys Glu
145 150 155 160
Ile Ile Phe Met Ile Asn Gly Lys Gly Ala Tyr Ser Lys Leu Lys Phe
165 170 175
Glu Asn Gly Ala His Arg Val Gln Arg Val Pro Glu Thr Glu Ser Gly
180 185 190
Gly Arg Ile His Thr Ser Thr Ala Thr Val Ala Cys Leu Pro Glu Met
195 200 205
Glu Glu Ile Glu Val Glu Ile Asn Glu Lys Asp Ile Arg Val Asp Thr
210 215 220
Phe Ala Ser Ser Gly Pro Gly Gly Gln Ser Val Asn Thr Thr Met Ser
225 230 235 240
Ala Val Arg Leu Thr His Ile Pro Thr Gly Ile Val Val Thr Cys Gln
245 250 255
Asp Glu Lys Ser Gln Ile Lys Asn Lys Glu Lys Ala Met Lys Val Leu
260 265 270
Arg Ala Arg Ile Tyr Asp Lys Tyr Gln Gln Glu Ala Arg Ala Glu Tyr
275 280 285
Asp Gln Thr Arg Lys Gln Ala Val Gly Thr Gly Asp Arg Ser Glu Arg
290 295 300
Ile Arg Thr Tyr Asn Phe Pro Gln Asn Arg Val Thr Asp His Arg Ile
305 310 315 320
Gly Leu Thr Ile Gln Lys Leu Asp Gln Val Pro Asp Gly His Leu Asp
325 330 335
Glu Ile Ile Glu Ala Leu Ile Leu Asp Asp Gln Ala Lys Lys Leu Glu
340 345 350
Gln Ala Asn Asp Ala Ser
355
<210> 134
<211> 255
<212> PRT
<213> Aequorea victoria
<400> 134
Met Arg Gly Ser His His His His His His Gly Ser Ser Lys Gly Glu
1 5 10 15
Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp
20 25 30
Val Asn Gly His Lys Phe Ser Val Arg Gly Glu Gly Glu Gly Asp Ala
35 40 45
Thr Asn Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu
50 55 60
Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Leu
65 70 75 80
Cys Phe Ser Arg Tyr Pro Asp His Met Lys Arg His Asp Phe Phe Lys
85 90 95
Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Ser Phe Lys
100 105 110
Asp Asp Gly Thr Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp
115 120 125
Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp
130 135 140
Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Phe Asn Ser His Asn
145 150 155 160
Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Tyr Phe
165 170 175
Lys Ile Arg His Asn Val Glu Asp Gly Ser Val Gln Leu Ala Asp His
180 185 190
Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp
195 200 205
Asn His Tyr Leu Ser Thr Gln Ser Val Leu Ser Lys Asp Pro Asn Glu
210 215 220
Lys Arg Asp His Met Val Leu Leu Glu Asp Val Thr Ala Ala Gly Ile
225 230 235 240
Thr His Gly Met Asp Glu Leu Tyr Lys Gly Ser Glu Pro Glu Ala
245 250 255
<210> 135
<211> 225
<212> PRT
<213> Aequorea victoria
<400> 135
Met Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly
1 5 10 15
Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp
20 25 30
Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys
35 40 45
Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val
50 55 60
Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe
65 70 75 80
Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe
85 90 95
Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly
100 105 110
Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu
115 120 125
Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His
130 135 140
Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn
145 150 155 160
Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp
165 170 175
His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro
180 185 190
Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn
195 200 205
Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly
210 215 220
Ile
225
<210> 136
<211> 883
<212> PRT
<213> T7 Bacteriophage
<400> 136
Met Asn Thr Ile Asn Ile Ala Lys Asn Asp Phe Ser Asp Ile Glu Leu
1 5 10 15
Ala Ala Ile Pro Phe Asn Thr Leu Ala Asp His Tyr Gly Glu Arg Leu
20 25 30
Ala Arg Glu Gln Leu Ala Leu Glu His Glu Ser Tyr Glu Met Gly Glu
35 40 45
Ala Arg Phe Arg Lys Met Phe Glu Arg Gln Leu Lys Ala Gly Glu Val
50 55 60
Ala Asp Asn Ala Ala Ala Lys Pro Leu Ile Thr Thr Leu Leu Pro Lys
65 70 75 80
Met Ile Ala Arg Ile Asn Asp Trp Phe Glu Glu Val Lys Ala Lys Arg
85 90 95
Gly Lys Arg Pro Thr Ala Phe Gln Phe Leu Gln Glu Ile Lys Pro Glu
100 105 110
Ala Val Ala Tyr Ile Thr Ile Lys Thr Thr Leu Ala Cys Leu Thr Ser
115 120 125
Ala Asp Asn Thr Thr Val Gln Ala Val Ala Ser Ala Ile Gly Arg Ala
130 135 140
Ile Glu Asp Glu Ala Arg Phe Gly Arg Ile Arg Asp Leu Glu Ala Lys
145 150 155 160
His Phe Lys Lys Asn Val Glu Glu Gln Leu Asn Lys Arg Val Gly His
165 170 175
Val Tyr Lys Lys Ala Phe Met Gln Val Val Glu Ala Asp Met Leu Ser
180 185 190
Lys Gly Leu Leu Gly Gly Glu Ala Trp Ser Ser Trp His Lys Glu Asp
195 200 205
Ser Ile His Val Gly Val Arg Cys Ile Glu Met Leu Ile Glu Ser Thr
210 215 220
Gly Met Val Ser Leu His Arg Gln Asn Ala Gly Val Val Gly Gln Asp
225 230 235 240
Ser Glu Thr Ile Glu Leu Ala Pro Glu Tyr Ala Glu Ala Ile Ala Thr
245 250 255
Arg Ala Gly Ala Leu Ala Gly Ile Ser Pro Met Phe Gln Pro Cys Val
260 265 270
Val Pro Pro Lys Pro Trp Thr Gly Ile Thr Gly Gly Gly Tyr Trp Ala
275 280 285
Asn Gly Arg Arg Pro Leu Ala Leu Val Arg Thr His Ser Lys Lys Ala
290 295 300
Leu Met Arg Tyr Glu Asp Val Tyr Met Pro Glu Val Tyr Lys Ala Ile
305 310 315 320
Asn Ile Ala Gln Asn Thr Ala Trp Lys Ile Asn Lys Lys Val Leu Ala
325 330 335
Val Ala Asn Val Ile Thr Lys Trp Lys His Cys Pro Val Glu Asp Ile
340 345 350
Pro Ala Ile Glu Arg Glu Glu Leu Pro Met Lys Pro Glu Asp Ile Asp
355 360 365
Met Asn Pro Glu Ala Leu Thr Ala Trp Lys Arg Ala Ala Ala Ala Val
370 375 380
Tyr Arg Lys Asp Lys Ala Arg Lys Ser Arg Arg Ile Ser Leu Glu Phe
385 390 395 400
Met Leu Glu Gln Ala Asn Lys Phe Ala Asn His Lys Ala Ile Trp Phe
405 410 415
Pro Tyr Asn Met Asp Trp Arg Gly Arg Val Tyr Ala Val Ser Met Phe
420 425 430
Asn Pro Gln Gly Asn Asp Met Thr Lys Gly Leu Leu Thr Leu Ala Lys
435 440 445
Gly Lys Pro Ile Gly Lys Glu Gly Tyr Tyr Trp Leu Lys Ile His Gly
450 455 460
Ala Asn Cys Ala Gly Val Asp Lys Val Pro Phe Pro Glu Arg Ile Lys
465 470 475 480
Phe Ile Glu Glu Asn His Glu Asn Ile Met Ala Cys Ala Lys Ser Pro
485 490 495
Leu Glu Asn Thr Trp Trp Ala Glu Gln Asp Ser Pro Phe Cys Phe Leu
500 505 510
Ala Phe Cys Phe Glu Tyr Ala Gly Val Gln His His Gly Leu Ser Tyr
515 520 525
Asn Cys Ser Leu Pro Leu Ala Phe Asp Gly Ser Cys Ser Gly Ile Gln
530 535 540
His Phe Ser Ala Met Leu Arg Asp Glu Val Gly Gly Arg Ala Val Asn
545 550 555 560
Leu Leu Pro Ser Glu Thr Val Gln Asp Ile Tyr Gly Ile Val Ala Lys
565 570 575
Lys Val Asn Glu Ile Leu Gln Ala Asp Ala Ile Asn Gly Thr Asp Asn
580 585 590
Glu Val Val Thr Val Thr Asp Glu Asn Thr Gly Glu Ile Ser Glu Lys
595 600 605
Val Lys Leu Gly Thr Lys Ala Leu Ala Gly Gln Trp Leu Ala Tyr Gly
610 615 620
Val Thr Arg Ser Val Thr Lys Arg Ser Val Met Thr Leu Ala Tyr Gly
625 630 635 640
Ser Lys Glu Phe Gly Phe Arg Gln Gln Val Leu Glu Asp Thr Ile Gln
645 650 655
Pro Ala Ile Asp Ser Gly Lys Gly Leu Met Phe Thr Gln Pro Asn Gln
660 665 670
Ala Ala Gly Tyr Met Ala Lys Leu Ile Trp Glu Ser Val Ser Val Thr
675 680 685
Val Val Ala Ala Val Glu Ala Met Asn Trp Leu Lys Ser Ala Ala Lys
690 695 700
Leu Leu Ala Ala Glu Val Lys Asp Lys Lys Thr Gly Glu Ile Leu Arg
705 710 715 720
Lys Arg Cys Ala Val His Trp Val Thr Pro Asp Gly Phe Pro Val Trp
725 730 735
Gln Glu Tyr Lys Lys Pro Ile Gln Thr Arg Leu Asn Leu Met Phe Leu
740 745 750
Gly Gln Phe Arg Leu Gln Pro Thr Ile Asn Thr Asn Lys Asp Ser Glu
755 760 765
Ile Asp Ala His Lys Gln Glu Ser Gly Ile Ala Pro Asn Phe Val His
770 775 780
Ser Gln Asp Gly Ser His Leu Arg Lys Thr Val Val Trp Ala His Glu
785 790 795 800
Lys Tyr Gly Ile Glu Ser Phe Ala Leu Ile His Asp Ser Phe Gly Thr
805 810 815
Ile Pro Ala Asp Ala Ala Asn Leu Phe Lys Ala Val Arg Glu Thr Met
820 825 830
Val Asp Thr Tyr Glu Ser Cys Asp Val Leu Ala Asp Phe Tyr Asp Gln
835 840 845
Phe Ala Asp Gln Leu His Glu Ser Gln Leu Asp Lys Met Pro Ala Leu
850 855 860
Pro Ala Lys Gly Asn Leu Asn Leu Arg Asp Ile Leu Glu Ser Asp Phe
865 870 875 880
Ala Phe Ala

Claims (61)

1.一种重组无细胞表达系统，包含：
-一种至少包含以下组分的核心重组蛋白混合物：
-多个起始因子(IF)；
-多个延伸因子(EF)；
-多个肽释放因子(RF)；
-至少一个核糖体循环因子(RRF)；
-多个氨酰-tRNA合成酶(RS)；以及
-至少一个甲硫氨酰-tRNA转甲酰基酶(MTF)；
-至少一个核酸合成模板；
-一种包含体外大分子合成所需的无细胞反应组分的反应混合物；以及
-其中，上述组分位于一个可进行大分子无细胞表达的生物反应器中。
2.根据权利要求1所述的系统，其中，所述核心重组蛋白混合物包括一种由细菌衍生的核心重组蛋白混合物。
3.根据权利要求2所述的系统，其中，所述由细菌衍生的核心重组蛋白混合物包括一种核心重组蛋白混合物，其中，至少一种组分由嗜热菌衍生而来。
4.根据权利要求2和3中任一项所述的系统，其中，所述嗜热菌包括芽孢杆菌嗜热菌或地芽孢杆菌嗜热菌。
5.根据权利要求4所述的系统，其中，从包括以下几项的一组中选择所述地芽孢杆菌嗜热菌：地表地芽孢杆菌和嗜热脂肪地芽孢杆菌。
6.根据权利要求1所述的系统，其中，所述由细菌衍生的核心重组蛋白混合物包括一种核心重组蛋白混合物，其中，至少一种组分由非嗜热菌或非嗜热菌和嗜热菌的组合衍生而来。
7.根据权利要求6所述的系统，其中，所述非嗜热菌包括大肠杆菌。
8.根据权利要求1所述的系统，其中，所述多个起始因子(IF)包括由嗜热菌衍生的多个起始因子。
9.根据权利要求1和8中任一项所述的系统，其中，所述由嗜热菌衍生的多个起始因子包括IF1、IF2、IF3或其任何片段或变体。
10.根据权利要求1、8和9中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个起始因子：SEQ ID NO.2、4、6、70、72和74，或序列一致性至少达90％的一个序列。
11.根据权利要求1所述的系统，其中，所述多个延伸因子(EF)包括由嗜热菌衍生的多个延伸因子。
12.根据权利要求1和11中任一项所述的系统，其中，所述由嗜热菌衍生的多个延伸因子包括EF-G、EF-Tu、EF-Ts、EF-4、EF-P或其任何片段或变体。
13.根据权利要求1、11和12中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个延伸因子：SEQ ID NO.8、10、12、14、16、76、78、80、82和84，或序列一致性至少达90％的一个序列。
14.根据权利要求1所述的系统，其中，所述多个肽释放因子(RF)包括由嗜热菌或芽孢杆菌衍生的多个肽释放因子。
15.根据权利要求1和14中任一项所述的系统，其中，所述由嗜热菌衍生的多个肽释放因子包括RF1、RF2和RF3或其任何片段或变体。
16.根据权利要求1、14和15中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个肽释放因子：SEQ ID NO.18、20、22、86和88，或序列一致性至少达90％的一个序列。
17.根据权利要求1所述的系统，其中，所述核糖体循环因子(RRF)包括由嗜热菌衍生的核糖体循环因子。
18.根据权利要求1和17中任一项所述的系统，其中，所述核糖体循环因子由地芽孢杆菌衍生而来。
19.根据权利要求1、17和18中任一项所述的系统，其中，所述核糖体循环因子包括包含以下氨基酸序列的核糖体循环因子：SEQ ID NO.14和90，或序列一致性至少达90％的一个序列。
20.根据权利要求1所述的系统，其中，所述多个氨酰-tRNA合成酶(RS)包括由嗜热菌或大肠杆菌衍生的多个氨酰-tRNA合成酶。
21.根据权利要求1和20中任一项所述的系统，其中，所述多个氨酰-tRNA合成酶包括AlaRS、ArgRS、AsnRS、AspRS、CysRS、GlnRS、GluRS、GlyRS、HisRS、IleRS、LeuRS、LysRS、MetRS、PheRS(a)、PheRS(b)、ProRS、SerRS、ThrRS、TrpRS、TyrRS和ValRS或其任何片段或变体。
22.根据权利要求1、20和21中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个氨酰-tRNA合成酶：SEQ ID NO.26、28、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、94、96、98、100、102、104、106、108、110、112、114、116、118、120、122、124、126、128和130，或序列一致性至少达90％的一个序列。
23.根据权利要求1所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶(MTF)包括由嗜热菌衍生的甲硫氨酰-tRNA转甲酰基酶。
24.根据权利要求1和23中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶由地芽孢杆菌衍生而来。
25.根据权利要求1、23和24中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶包括包含以下氨基酸序列的甲硫氨酰-tRNA转甲酰基酶：SEQ ID NO.68和132，或序列一致性至少达90％的一个序列。
26.根据权利要求1所述的系统，其中，所述核酸合成模板包括一个DNA模板。
27.根据权利要求26所述的系统，其中，所述DNA模板包括一个线性DNA模板，其包含：
-至少一个可连接启动子的靶序列，其中，所述靶序列可进行密码子优化；
-至少一个核糖体结合位点(RBS)；
-至少一个表达产物酶切位点；以及
-至少一个标签。
28.根据权利要求1所述的系统，其中，所述核酸合成模板包括一个RNA模板。
29.根据权利要求1所述的系统，其中，所述反应混合物包含以下一种或多种组分：
-一定量的核糖体，以及一定量的由嗜热菌衍生的核糖体(可选)；
-一定量的RNase抑制剂；
-一定量的RNA聚合酶；
-一定量的tRNA，以及一定量的由嗜热菌衍生的tRNA(可选)；
-一种缓冲液；以及
-一定量的氨基酸。
30.根据权利要求29所述的系统，其中，所述反应混合物进一步包含以下一种或多种组分：
-Tris乙酸盐；
-Mg(OAc)2；
-K+谷氨酸；
-氨基乙酸盐；
-NaCl；
-KCl；
-MgCk；
-DTT；
-辛基-b-糖苷；
-NAD；
-NADP；
-山梨糖醇；
-FADH；
-CoA；
-PLP；以及
-SAM。
31.根据权利要求1和29中任一项所述的系统，进一步包含一个能量源。
32.根据权利要求32所述的系统，其中，所述能量源包括一定量的核苷三磷酸(NTP)。
33.根据权利要求32所述的系统，其中，所述核苷三磷酸包括从包含以下几项的一组中选择的一种或多种核苷三磷酸：腺嘌呤三磷酸(ATP)、鸟苷三磷酸(GTP)、尿苷三磷酸(UTP)和胞苷三磷酸(CTP)。
34.根据权利要求31、32和33中任一项所述的系统，其中，所述能量源包括一个无机多磷酸盐能量再生系统。
35.根据权利要求34所述的系统，其中，所述无机多磷酸盐能量再生系统包括：
-一个细胞三磷酸腺苷(ATP)能量再生系统，包含：
-一定量的腺苷激酶(GstAdK)；
-一定量的多磷酸盐激酶(Taq PPK)；
-一定量的无机多磷酸盐(PPi)；以及
-一定量的一磷酸腺苷(AMP)；
-其中，所述AdK酶和PPK酶发挥协同作用，从PPi和AMP中再生细胞能量。
36.根据权利要求1所述的系统，其中，所述生物反应器包括一个连续流动式生物反应器。
37.一种重组无细胞表达反应混合物，包含：
-多个起始因子(IF)；
-多个延伸因子(EF)；
-多个释放因子(RF)；
-至少一个核糖体循环因子(RRF)；
-多个氨酰-tRNA合成酶(RS)；以及
-至少一个甲硫氨酰-tRNA转甲酰基酶(MTF)。
38.根据权利要求37所述的系统，其中，所述多个起始因子(IF)包括由嗜热菌衍生的多个起始因子。
39.根据权利要求37和38中任一项所述的系统，其中，所述由嗜热菌衍生的多个起始因子包括IF1、IF2、IF3或其任何片段或变体。
40.根据权利要求37、38和39中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个起始因子：SEQ ID NO.2、4、6、70、72和74，或序列一致性至少达90％的一个序列。
41.根据权利要求37所述的系统，其中，所述多个延伸因子(EF)包括由嗜热菌衍生的多个延伸因子。
42.根据权利要求37和41中任一项所述的系统，其中，所述由嗜热菌衍生的多个延伸因子包括EF-G、EF-Tu、EF-Ts、EF-4、EF-P或其任何片段或变体。
43.根据权利要求37、41和42中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个延伸因子：SEQ ID NO.8、10、12、14、16、76、78、80、82和84，或序列一致性至少达90％的一个序列。
44.根据权利要求37所述的系统，其中，所述多个肽释放因子(RF)包括由嗜热菌或芽孢杆菌衍生的多个释放因子。
45.根据权利要求37和44中任一项所述的系统，其中，所述多个肽释放因子包括RF1、RF2和RF3或其任何片段或变体。
46.根据权利要求37、44和45中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个肽释放因子：SEQ ID NO.18、20、22、86和88，或序列一致性至少达90％的一个序列。
47.根据权利要求37所述的系统，其中，所述核糖体循环因子(RRF)包括由嗜热菌衍生的核糖体循环因子。
48.根据权利要求37和47中任一项所述的系统，其中，所述核糖体循环因子由地芽孢杆菌衍生而来。
49.根据权利要求37、47和48中任一项所述的系统，其中，所述核糖体循环因子包括包含以下氨基酸序列的核糖体循环因子：SEQ ID NO.14和90，或序列一致性至少达90％的一个序列。
50.根据权利要求37所述的系统，其中，所述多个氨酰-tRNA合成酶(RS)包括多个氨酰-tRNA合成酶，其中，至少一个氨酰-tRNA合成酶由嗜热菌衍生而来。
51.根据权利要求37和50中任一项所述的系统，其中，所述多个氨酰-tRNA合成酶包括AlaRS、ArgRS、AsnRS、AspRS、CysRS、GlnRS、GluRS、GlyRS、HisRS、IleRS、LeuRS、LysRS、MetRS、PheRS(a)、PheRS(b)、ProRS、SerRS、ThrRS、TrpRS、TyrRS和ValRS或其任何片段或变体。
52.根据权利要求37、50和51中任一项所述的系统，其中，从包括以下几项的一组氨基酸序列中选择所述多个氨酰-tRNA合成酶：SEQ ID NO.26、28、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、94、96、98、100、102、104、106、108、110、112、114、116、118、120、122、124、126、128和130，或序列一致性至少达90％的一个序列。
53.根据权利要求37所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶(MTF)包括由嗜热菌衍生的甲硫氨酰-tRNA转甲酰基酶。
54.根据权利要求37和53中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶由地芽孢杆菌衍生而来。
55.根据权利要求37、53和54中任一项所述的系统，其中，所述甲硫氨酰-tRNA转甲酰基酶包括包含以下氨基酸序列的甲硫氨酰-tRNA转甲酰基酶：SEQ ID NO.68和132，或序列一致性至少达90％的一个序列。
56.一种分离核苷酸，包括从包含以下几项的一组中选择的一种核苷酸：
-SEQ ID NO.1、3、5、69、71和73；
-SEQ ID NO.7、9、11、13、15、75、77、79、81和83；
-SEQ ID NO.17、19、21、85和87；
-SEQ ID NO.23和89；以及
-SEQ ID NO.25、27、29、31、33、35、37、39、41、43、45、47、49、51、53、55、57、59、61、63、65、67、91、93、95、97、99、101、103、105、107、109、111、113、115、117、119、121、123、125、127、129和131。
57.一种表达载体，包含至少一个权利要求56所述的核苷酸序列(可连接启动子)。
58.一种由权利要求57所述的一种表达载体转化的细菌。
59.根据权利要求58所述的转化细菌，其中，所述细菌包括大肠杆菌。
60.一种肽，包含从包括以下几项的一组中选择的一个氨基酸序列：
-SEQ ID NO.2、4、6、70、72和74；
-SEQ ID NO.8、10、12、14、16、76、78、80、82和84；
-SEQ ID NO.18、20、22、86、88；
-SEQ ID NO.14和90；
-SEQ ID NO.26、28、32、34、36、38、40、42、44、46、48、50、52、54、56、58、60、62、64、66、94、96，SEQ ID NO.98、100、102、104、106、108、110、112、114、116、118、120、122、124、126、128和130；以及
-SEQ ID NO.68和132，或其任何片段或变体。
61.一种无细胞表达系统，其使用权利要求60所述的至少一种肽。
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