CN108779444A - 生产脂肪酸的方法 - Google Patents
生产脂肪酸的方法 Download PDFInfo
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Abstract
本发明涉及通过在宿主细胞中表达包含异源硫酯酶(TE)的天然缺乏硫酯酶(TE)结构域的重组脂肪酸合酶(FAS)来生产脂肪酸，特别是短/中链脂肪酸(S/MCFA)的方法。本发明进一步涉及包含此类重组FAS的多核苷酸、由此编码的多肽和包含此类多核苷酸的载体，以及包含所述多核苷酸、多肽或载体的重组宿主细胞。
Description
发明领域
本发明涉及脂肪酸生产，特别是短/中链脂肪酸(S/MCFA)和衍生产物例如碳氢化合物、脂肪醛和脂肪醇的生产的领域。本发明至此提供了遗传构建体和宿主细胞，生产这些的方法以及使用这些的方法。
背景技术
短/中链脂肪酸(S/MCFA，例如C6-C12)是许多工业化学品和生物燃料的前体。S/MCFA的主要来源是来自植物油和化石油的精炼，但这些化合物的世界可用性有限，而消耗量却在增加。因此，特别感兴趣的是找到这些产品的新来源，并同时通过微生物转化生物质基材料以可持续的方式生产这些种类的脂肪酸。
微生物脂肪酸合成的天然过程通过将两个碳单元逐步添加到与酰基载体蛋白(ACP)结合的生长酰基链上行进。该过程开始于乙酰-ACP和丙二酰-ACP缩合成乙酰乙酰-ACP，释放出驱动反应向前的CO2。第二个步骤牵涉使用NADPH将乙酰乙酰-ACP还原为D-3-羟丁酰-ACP。在脱水成巴豆酰-ACP并使用NADPH进行另一次还原后，形成丁酰-ACP。链延伸通常继续进一步添加丙二酰-ACP直至形成一定长度的酰基链，其然后通过硫酯酶水解成游离脂肪酸。
最近，对于链长控制的大多数努力集中在脂肪酸合成或β-氧化的逆转上，并且限制酰基链延伸的靶标是负责缩合(酮脂酰-ACP合酶和硫解酶)或产物释放(酰基-ACP/CoA硫酯酶)的酶。
在大肠杆菌中首先建立了β-氧化的逆转，并鉴定了该途径的各个功能部分。然而，该途径优选产生短链羧酸(丁酸)。进一步的工程牵涉使用能够将乙酰辅酶A与C4-C8酰基辅酶A中间体缩合以产生中链脂肪酸的另一种硫解酶(例如BktB)。大肠杆菌中的滴度超过1g/L(C6-C10)。最近，在测试了超过40种酶后，在酵母中实现了逆转的β-氧化，尽管其生产能力远低于大肠杆菌。
在原核生物中，使用解离的II型脂肪酸合酶。脂肪酸合成的终产物为酰基-ACP。生产S/MCFA的常见策略是表达短链酰基-ACP硫酯酶，其从ACP硫代酸酯中释放S/MCFA。短/中链酰基-ACP，硫酯酶的底物，也掺入长链脂肪酸合成中。因此使用对中/长链酰基-ACP(C8-C14)具有较低亲和力的浅蓝菌素抗性β-酮脂酰-ACP合酶突变体(FabF*)以阻断短/中链酰基-ACP的掺入。在大肠杆菌中使用的组合策略产生118mg/LC8脂肪酸的滴度。
从上文可以清楚地看出，为了生产脂肪酸例如S/MCFA，研究并采用了各种策略，特别是脂肪酸的微生物生产。然而，本领域仍然需要新的和改进的脂肪酸生产，例如S/MCFA的方法。本发明至此提供了解决方案。
发明内容
本发明涉及生产或牵涉生产脂肪酸的方法和手段。本发明特别涉及用于生产脂肪酸例如短和中链脂肪酸的生物体(例如酵母，藻类......)的工程化改造。根据本发明，通过导入硫酯酶(TE)来工程化改造脂肪酸合酶(FAS)例如优选真菌FAS或它的亚基，从而导致脂肪酸，特别是短或中链脂肪酸(S/MCFA)的产生。本发明集中在经修饰的真菌FAS(亚基)与TE，特别是异源TE的特定组合上，以特别是获得具有定制碳链长度(例如C6至C12)的脂肪酸。
根据本发明，用硫酯酶结构域补充，或者通过优选在酰基载体蛋白(ACP)之前或之后(即邻近ACP结构域)添加硫酯酶(TE)结构域来工程化改造没有硫酯酶(TE)结构域的脂肪酸合酶(FAS)，例如优选真菌FAS或它的亚基，从而导致脂肪酸，特别是短或中链脂肪酸(S/MCFA)的产生。
本发明的基础是经修饰的真菌脂肪酸合酶(FAS)或它的亚基的使用。本发明令人惊讶地发现，用TE补充FAS，或者例如在现有的ACP结构域之前或之后将TE结构域添加到缺乏此类TE结构域的FAS(亚基)，允许调整(tailor)脂肪酸长度，而不会以其他方式影响或干扰它的脂肪酸合成功能。该发现对于I型FAS更加令人惊讶，所述I型FAS中各个酶结构域作为单个(或作为两个亚基)大的多功能多肽编码。各个FAS结构域共价连接并因此在结构上受到约束。出乎意料地，在这样的背景下，导入另一个结构域(即TE)不仅不扰乱脂肪酸合成，而且还允许调整脂肪酸长度。此外，已经发现一些真菌FAS(亚基)基因编码多个ACP结构域。有利地，本发明人已经发现，不仅删除或使这些重复的ACP结构域之一失活而不丧失功能是可能的，而且另外地可以通过用TE结构域替换一个此类ACP结构域来改变功能是可能的。根据本发明的此类经修饰的FAS(亚基)的维持的功能更加令人惊讶。通常，ACP位于FAS中内部，使得不能预期提供另外的TE例如直接在ACP的上游或下游而不改变或特别是恶化FAS(亚基)结构和活性。
通过所附权利要求特别地体现了本发明，所附权利要求在此通过引用明确地并入本文。
一方面，本发明涉及编码脂肪酸合酶(FAS)或FAS亚基的重组多核酸，特别是真菌的，特别是酵母、原生生物、黏菌或藻类，特别是微藻FAS，以及硫酯酶(TE)，在某些优选实施方案中，其作为单一ORF编码，和/或其中相对于所述FAS(亚基)的酰基载体蛋白(ACP)直接5'或3'提供所述TE。
另一方面，本发明涉及重组原核或真核细胞，所述细胞包含、表达或能够表达，例如诱导地或组成地表达，脂肪酸合酶(FAS)或FAS亚基(例如FAS1和/或FAS2)，特别是真菌的，特别是酵母、原生生物、黏菌或藻类，特别是微藻FAS，以及(异源的)硫酯酶(TE)，在某些实施方案中，其作为单一ORF编码，和/或其中相对于所述FAS(亚基)的酰基载体蛋白(ACP)直接5'或3'(或者N-末端或C-末端)提供所述TE。
应当理解的是，根据本发明，天然存在的FAS(亚基)缺乏TE，并因此可以将FAS(亚基)工程化改造以另外包含或含有TE，或者用TE补充FAS，使得例如如本文所述的根据本发明的重组细胞包含、表达或能够表达FAS和TE。
因此，一方面，本发明涉及包含、表达或能够表达脂肪酸合酶(FAS)或FAS亚基基因序列的重组原核或真核细胞，或者包含脂肪酸合酶(FAS)或FAS亚基基因序列的多核酸，所述基因序列源自缺乏硫酯酶结构域(TE)编码核苷酸序列的天然存在的FAS(亚基)基因，其中将异源硫酯酶结构域(TE)编码核苷酸序列添加到所述FAS(亚基)，优选在酰基载体蛋白结构域(ACP)编码核苷酸序列之前或之后(即5’或3’)。
另一方面，本发明涉及包含、表达或能够表达脂肪酸合酶(FAS)或FAS亚基基因序列的重组原核或真核细胞，或者包含脂肪酸合酶(FAS)或FAS亚基基因序列的多核酸，所述基因序列源自具有至少两个酰基载体蛋白(ACP)编码核苷酸序列的FAS(亚基)基因，例如天然存在的FAS(亚基)基因，例如真菌FAS(亚基)特别是酵母、原生生物、黏菌或藻类，特别是微藻FAS(亚基)基因，其中用硫酯酶(TE)编码核苷酸序列替换所述ACP编码核苷酸序列之一。
相关方面的本发明还关注由本文定义的重组多核酸编码的多肽，以及包含本文定义的多核酸的载体和宿主细胞，例如原核或真核宿主细胞。
另一方面，本发明涉及包含FAS(亚基)基因序列的重组原核或真核宿主细胞，所述基因序列源自真菌FAS(亚基)基因并另外编码TE编码核苷酸序列。因此，本发明还涉及包含FAS(亚基)基因序列的重组宿主细胞，所述基因序列源自缺乏硫酯酶(TE)结构域编码核苷酸序列的天然存在的FAS(亚基)基因，其中将异源硫酯酶(TE)结构域编码核苷酸序列添加到所述FAS(亚基)，优选在酰基载体蛋白结构域(ACP)编码核苷酸序列之前或之后(即5’或3’)。在另一个实施方案中，所述TE未整合到所述FAS(亚基)编码核苷酸序列中。因此，在某些实施方案中，本发明涉及包含FAS(亚基)基因序列的重组原核或真核宿主细胞，所述基因序列源自真菌FAS(亚基)基因并另外编码TE编码核苷酸序列，其中所述TE编码核酸序列位于与所述FAS编码核酸序列不同的基因座。在某些实施方案中，所述FAS和/或所述TE是基因组整合的。在某些实施方案中，所述FAS和/或所述TE是附加型的，例如在载体上提供。
在相关方面，本发明关注生产如本文所述的重组原核或真核细胞的方法。在某些实施方案中，所述方法包含在原核或真核细胞中导入如本文所述的重组多核酸、多肽或载体的步骤，以及此类细胞用于生产脂肪酸和它们的衍生物，例如碳氢化合物和脂肪醇的用途，和生产脂肪酸、牵涉此类细胞的使用的相关方法。
在衍生的进一步方面，本发明还涉及生产碳氢化合物，例如烷烃或烯烃，或脂肪醇或脂肪醛的方法，牵涉根据如本文所述的方法生产的脂肪酸的同时或下游转化，特别牵涉脂肪酸的还原、氢化、脱羧或脱碳。
附图简述
图1真菌I型FAS的结构域构造。在真菌I型FAS中有八个不同的蛋白质结构域，即乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰/棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)。与来自酿酒酵母的FAS不同，来自圆红冬孢酵母或Aplanochytrium kerguelense的FAS含有两个串联的ACP结构域，并用硫酯酶(TE)结构域替换第一或第二个ACP结构域中的任一个。在来自酿酒酵母的FAS中，将此类异源酰基-ACP硫酯酶(TE)结构域插入到ACP结构域的上游或下游区域中。这些种类的含有TE结构域的经修饰的FAS用于生产经调整的中链脂肪酸。
图2来自圆红冬孢酵母(A)和Aplanochytrium kerguelense(B)和酿酒酵母(C)的FAS的表达组件。在RtFAS2-ACPI2TE、RtFAS2-ACPII2TE AkFAS2-ACPI2TE和AkFAS2-ACPII2TE中，用来自Acinetobacter baylyi的短链酰基-ACP硫酯酶替换任一ACP结构域。在ScFAS2-TE-ACP和ScFAS2-ACP-TE中，将来自A.baylyi的短链酰基-ACP硫酯酶插入到ACP结构域上游或下游的ScFAS2中。
图3RtFAS与酿酒酵母PWY12(MATαura3 leu2 his3 trp1 can1 Δfas1::HIS3 Δfas2::LEU2)的脂肪酸营养缺陷型的互补。
图4由RtFAS和它的突变体生产的中链脂肪酸(己酸，C6；辛酸，C8；癸酸，C10；和十二烷酸，C12)。庚酸(C7)用作内标。
图5由表达相应的FAS的酿酒酵母PWY12生产的细胞外脂肪酸的滴度，所述FAS包括来自酿酒酵母的ScFAS和来自圆红冬孢酵母的FAS(野生型，WT；FAS/TE杂合体，ACPI2TE和ACPII2TE)。
图6表达相应的FAS的酿酒酵母PWY12的细胞内脂肪酸组成，所述FAS包括来自酿酒酵母的ScFAS和来自圆红冬孢酵母的FAS(野生型，WT；FAS/TE杂合体，ACPI2TE和ACPII2TE)。
图7由表达相应的FAS的酿酒酵母PWY12生产的细胞外脂肪酸的滴度，所述FAS来自Aplanochytrium kerguelense(野生型，AkFAS-WT；FAS/TE杂合体，AkFAS-ACPI2TE和AkFAS-ACPII2TE)。
图8由表达相应的FAS的酿酒酵母PWY12生产的细胞外脂肪酸的滴度，所述FAS来自酿酒酵母(野生型，ScFAS-WT；FAS/TE杂合体，ScFAS-TE-ACP和ScFAS-ACP-TE)。
图9由整合地表达相应的FAS的酿酒酵母YJZ02生产的细胞外脂肪酸的滴度，所述FAS来自酿酒酵母(野生型，WT；FAS/TE杂合体，ACP-TE)。
图10适用于根据本发明的实施方案的序列。
图11用于工程化改造真菌脂肪酸合酶(FAS)以合成经调整的油脂化学品的设计和策略。(a)真菌FAS变体的结构域组成。(b)真菌FAS结构的横截面展示。在上部区室中仅对于三组活性位点之一显示每个结构域的位置。可以如图所示将异源的酶(黑色圆圈)插入到反应室中。(c)由经工程化改造的真菌FAS催化的反应循环。将异源短链硫酯酶(sTE)整合到延伸循环中以释放短/中链脂肪酸。AT，乙酰转移酶；ER，烯酰还原酶；DH，脱水酶；MPT，丙二酰-棕榈酰转移酶；ACP，酰基载体蛋白；KS，酮脂酰合酶；KR，酮脂酰还原酶；PPT，磷酸泛酰巯基乙胺基转移酶。
图12通过同源PPT对来自圆红冬孢酵母脂肪酸合酶的ACP结构域进行体外磷酸泛酰巯基乙胺化(phosphopantetheinylation)。(a)由磷酸泛酰巯基乙胺基转移酶(PPT)催化的磷酸泛酰巯基乙胺化反应。将辅酶A(CoA)的磷酸泛酰巯基乙胺部分转移至酰基载体蛋白(ACP)的丝氨酸残基的羟基。(b)磷酸泛酰巯基乙胺化产物的SDS-PAGE分析。作为GST融合蛋白表达和纯化wACP I(RtFas2，1022-1184)、wACPII(RtFas2，1213-1375)和PPT(RtFas2，2809-2928)。在mACPI和mACPII中，相应的丝氨酸突变为丙氨酸，如(b)中所示。通过肠激酶切割磷酸泛酰巯基乙胺化反应混合物，并通过16％Tricine-SDS-PAGE分离。箭头显示了分子量略高于apo-ACP的经磷酸泛酰巯基乙胺化的ACP。
图13经纯化的RtFAS复合物的体外酶活性测定。(a)经纯化的RtFAS复合物的SDS-PAGE分析，如图所示两个亚基是分开的。(b)经纯化的RtFAS和它的突变体的酶活性测定。通过340nm处吸光度的减少来监测丙二酰辅酶A和乙酰辅酶A依赖性NADPH氧化。
图14通过不同的FAS和它们的突变体对酿酒酵母PWY12中FAS1和FAS2缺失的互补。(a)在含有脂肪酸(SC-Ura+FA)或不含脂肪酸(SC-Ura)的平板上具有相应质粒的PWY12的生长。具有空白载体(pYX212)的菌株只能在补充脂肪酸的平板上生长。(b)如图所示在具有100mg/L色氨酸的Delft培养基中的具有质粒的PWY12菌株的生长曲线。将细胞以200μl的体积培养(在蜂窝板中)，并通过Bioscreen C MBR仪器监测光密度。(c)-(f)，如图所示在不含脂肪酸(SC-Ura)的平板上的具有质粒的PWY12的生长。
图15通过经工程化改造的真菌FAS合成油脂化学品。(a)经工程化改造的真菌FAS的示意图。ACP的保守丝氨酸残基突变为丙氨酸残基以使ACP结构域失活。在ScFAS27和ScFAS28中，将G1250S，S1251W突变导入到ScFas2蛋白的酮脂酰-ACP合酶(KS)结构域中。在经工程化改造的ScFAS中，分离RtFAS中两个串联ACP的柔性接头用于分离异源蛋白质和ACP结构域。(b-d)如图所示通过表达真菌FAS的菌株酿酒酵母PWY12的细胞外脂肪酸的生产。提取并量化己酸(C6)、辛酸(C8)、癸酸(C10)、十二烷酸(C12)。超过3个独立培养物用于产物量化。显示了平均值±SD。*，p<0.01以及**，p<0.001。Student’s t检验。
图16具有嵌入的(ScFAS14和ScFAS15)、外围的(ScFAS16)或游离的(ScFAS17)sTE的ScFAS的示意图。
图17在具有经工程化改造的ScFAS整合的YJZ02中的S/MCFA的生产。(a)在具有表达AcTesA的质粒的ZW201菌株(具有整合了ScFAS01的染色体的YJZ02)和具有空载体的ZW206菌株(具有整合了ScFAS15的染色体的YJZ02)中的S/MCFA生产。在补充有100mg/L组氨酸的Delft培养基中培养细胞48小时。(b)菌株ZW201、ZW206、ZW202(具有整合了ScFAS27的染色体的YJZ02)和ZW207(具有整合了ScFAS28的染色体的YJZ02)的S/MCFA生产和光密度(OD)。在补充有100mg/L组氨酸和100mg/L尿嘧啶的Delft培养基中培养细胞48、72和96小时。(c)在补充有100mg/L组氨酸和100mg/L尿嘧啶的Delft培养基中的ZW207的调节pH的培养。箭头显示添加无菌KOH溶液(2M)以将pH改变为6.0。(d)如图(c)中所示，在未调节pH和调节pH的培养期间ZW207的S/MCFA生产。显示了3-4个独立培养物的平均值±SD。
发明详述
将结合特定实施方案描述本发明，但是本发明不限于此。
本发明的组合物、方法和用途的优选陈述(特征)和实施方案在下文中列出。除非明确地表明相反，否则如此定义的本发明的每个陈述和实施方案可以与任何其他陈述和/或实施方案组合。特别地，表明为优选或有利的任何特征可以与表明为优选或有利的任何其他特征或多个特征或陈述组合。至此，特别地通过一个或多个以下编号方面和实施方案1至68中的任何一个或与任何其他陈述和/或实施方案的任何组合来体现本发明。
本申请中公开的编号陈述是：
1.重组多核酸，其编码(i)脂肪酸合酶(FAS)或FAS亚基；和(ii)硫酯酶(TE)，优选异源TE，优选重组多核酸，其包含(i)脂肪酸合酶(FAS)编码序列或FAS亚基编码序列，或部分脂肪酸合酶(FAS)编码序列，源自或衍生自缺乏硫酯酶(TE)编码核苷酸序列的天然存在的FAS(亚基)基因，其中将异源硫酯酶结构域(TE)编码核苷酸序列添加到所述FAS(亚基)，优选在所述FAS(亚基)的酰基载体蛋白结构域(ACP)编码核苷酸序列之前或之后，任选地其中所述FAS亚基为FAS1或FAS2或1。重组原核或真核细胞包含、表达或能够表达，或重组多核酸编码(i)脂肪酸合酶(FAS)或FAS亚基；和(ii)硫酯酶(TE)，优选异源TE，优选重组原核或真核细胞包含、表达或能够表达，或重组多核酸包含(i)脂肪酸合酶(FAS)编码序列或FAS亚基编码序列，或部分脂肪酸合酶(FAS)编码序列，源自或衍生自缺乏硫酯酶(TE)编码核苷酸序列的天然存在的FAS(亚基)基因，其中将异源硫酯酶结构域(TE)编码核苷酸序列添加到所述FAS(亚基)，优选在所述FAS(亚基)的酰基载体蛋白结构域(ACP)编码核苷酸序列之前或之后，任选地其中所述FAS亚基为FAS1或FAS2。
2.根据陈述1的细胞或多核酸，其中所述FAS(亚基)和所述TE作为单一开放阅读框(ORF)编码。
3.根据陈述1或2的细胞或多核酸，其中编码所述TE的多核酸序列相对于编码所述FAS(亚基)的酰基载体蛋白(ACP)的多核酸序列相邻(直接5’或3’)。
4.根据陈述1至3中任一项的细胞或多核酸，包含脂肪酸合酶(FAS)编码序列或FAS亚基编码序列，或部分脂肪酸合酶(FAS)编码序列，源自或衍生自具有至少两个酰基载体蛋白(ACP)编码核苷酸序列的FAS(亚基)基因，例如天然存在的FAS(亚基)基因，其中用硫酯酶(TE)编码核苷酸序列替换所述ACP编码核苷酸序列之一。
5.根据陈述1至4中任一项的细胞或多核酸，其中所述天然存在的FAS(亚基)基因为I型FAS(亚基)。
6.根据陈述1至5中任一项的细胞或多核酸，其中所述FAS(亚基)是真菌的，特别是酵母、原生生物、黏菌或藻类，特别是微藻FAS(亚基)。
7.根据陈述1至6中任一项的细胞或多核酸，其中所述FAS(亚基)来自酵母属，优选酿酒酵母，红冬孢酵母属(Rhodosporidium spp.)，优选圆红冬孢酵母或Aplanochytriumspp.，优选A.kerguelense。
8.根据陈述1至7中任一项的细胞或多核酸，其中所述TE为酰基-CoA/ACP TE，优选短链或中链酰基-CoA/ACP TE。
9.由根据陈述1至8中任一项的重组多核酸编码的重组多肽。
10.包含根据陈述1至8中任一项的重组多核酸或根据陈述9的编码重组多肽的多核酸的重组载体。
11.根据陈述10的重组载体，其中所述载体为表达载体或重组载体。
12.重组原核或真核细胞，所述细胞包含根据陈述1至8中任一项的重组多核酸、根据陈述9的多肽、根据陈述6的编码多肽的多核酸或根据陈述10或11的载体。
13.根据陈述12的重组原核或真核细胞，其中所述重组多核酸是基因组整合的。
14.重组原核或真核细胞，所述细胞包含源自FAS(亚基)基因例如天然存在的FAS(亚基)基因的FAS(亚基)蛋白序列或FAS(亚基)蛋白编码序列，和TE蛋白序列或TE蛋白编码序列，其中任选地所述FAS(亚基)具有至少两个ACP多肽或ACP多肽编码核苷酸序列，其中用TE多肽或TE多肽编码核苷酸序列替换所述ACP多肽或ACP多肽编码核苷酸序列之一。
15.根据陈述1至14中任一项的重组原核或真核细胞，其中所述TE或TE编码核苷酸序列与天然存在的FAS(亚基)相比为异源TE编码核苷酸序列。
16.根据陈述1至15中任一项的重组原核或真核细胞，其为细菌细胞、真菌细胞或藻类细胞。
17.根据陈述1至16中任一项的重组原核或真核细胞，其为酵母属细胞，例如酿酒酵母细胞。
18.生产重组原核或真核细胞的方法，包含以下步骤：在原核或真核细胞中导入根据陈述1至11或27-56中任一项的重组多核酸、多肽或载体，或在原核或真核细胞中导入FAS，任选地FAS1和/或FAS2，和TE，或编码FAS，任选地FAS1和/或FAS2，和TE的多核酸。
19.生产重组原核或真核细胞的方法，包含以下步骤：
(i)提供原核或真核细胞，所述细胞包含脂肪酸合酶(FAS)基因或FAS亚基基因，任选地FAS1和/或FAS2，任选地所述FAS(亚基)具有至少两个酰基载体蛋白(ACP)编码核苷酸序列，例如包含两个相邻的ACP编码核苷酸序列，优选所述FAS(亚基)缺乏TE编码核苷酸序列；和
(ii)导入TE或TE编码序列，任选地用TE编码序列替换所述ACP编码核苷酸序列之一，其中优选导入所述TE编码核苷酸序列，优选直接地，所述FAS(亚基)的ACP编码核苷酸序列的5’或3’。
20.根据陈述1至17或27-67中任一项的重组多核酸、多肽、载体或细胞在脂肪酸、碳氢化合物、脂肪醛或脂肪醇的生产中的用途。
21.生产脂肪酸的方法，包含以下步骤：
(i)提供根据陈述1至7或60至67中任一项的重组原核或真核细胞；和
(ii)在适合的培养条件下培养所述重组原核或真核细胞。
22.根据陈述21的方法，进一步包含以下步骤：
(iii)分离和/或纯化所述脂肪酸。
23.生产(中或短链)碳氢化合物的方法，包含实施根据陈述21或22的生产脂肪酸的方法，进一步包含所述脂肪酸的还原、氢化、脱羧或脱碳。
24.生产(中或短链)脂肪醇或脂肪醛的方法，包含实施根据陈述21或22的生产脂肪酸的方法，进一步包含氢化所述脂肪酸，任选地在所述脂肪酸的酯化之后；或进一步包含还原所述脂肪酸。
25.根据陈述20的用途或根据陈述21至23的方法，其中所述脂肪酸为短链脂肪酸(SCFA)或中链脂肪酸(MCFA)。
26.根据陈述20的用途或根据陈述21至23中任一项的方法，其中所述脂肪酸为C1-C12脂肪酸，例如C6-C12脂肪酸。
27.根据陈述1至17或49至56中任一项的重组原核或真核细胞或重组多核酸，其中所述FAS由一个或两个分立的和/或分离的基因序列编码。
28.根据陈述1至17或27或49至56中任一项的重组原核或真核细胞或重组多核酸，其中所述FAS是或包含FAS1和/或FAS2。
29.根据陈述1至8或27至28或49至56中任一项的重组原核或真核细胞或重组多核酸，其中所述FAS是或包含FAS亚基α和/或FAS亚基β。
30.根据陈述1至8或27至28或49至56中任一项的重组原核或真核细胞或重组多核酸，其中所述FAS(亚基)源于表1中列出的生物体，优选酵母属(例如酿酒酵母)、红冬孢酵母属(例如圆红冬孢酵母)或Aplanochytrium sp.(例如A.kerguelense)。
31.根据陈述1至8或27至28或49至56中任一项的重组原核或真核细胞或重组多核酸，其中所述FAS(亚基)包含或由以下组成：如SEQ ID NO:1至4或19中所阐明的序列、它们的片段，或者与SEQ ID NO:1至4或19具有至少50％、优选至少60％、更优选至少70％、最优选至少80％序列同一性的序列，或它们的片段和/或它们的直系同源物。
32.根据陈述31的重组原核或真核细胞或重组多核酸，其中所述片段包含ACP编码序列。
33.根据陈述31的重组原核或真核细胞或重组多核酸，其中与SEQ ID NO:1至4或19具有至少50％、优选至少60％、更优选至少70％、最优选至少80％序列同一性的所述序列为包含或由如SEQ ID NO:1至4或19中所列的序列组成的所述FAS(亚基)的直系同源物或功能变体。
34.根据陈述1至8或27至30或49至56中任一项的重组原核或真核细胞或重组多核酸，包含多核酸序列，所述多核酸序列包含或由以下组成：如SEQ ID NO:5、6、15或16中所阐明的序列、它们的片段，或者与SEQ ID NO:5、6、15或16具有至少50％、优选至少60％、更优选至少70％、最优选至少80％序列同一性的序列，或它们的片段和/或它们的直系同源物。
35.根据陈述34的重组原核或真核细胞或重组多核酸，其中所述片段包含ACP编码核苷酸序列和TE编码核苷酸序列，优选从5’至3’为ACP编码核苷酸序列和TE编码核苷酸序列，或从5’至3’为TE编码核苷酸序列和ACP编码核苷酸序列，优选不含居间的(酶)编码核苷酸序列。
36.根据陈述34的重组原核或真核细胞或重组多核酸，其中与SEQ ID NO:5、6、15或16具有至少50％、优选至少60％、更优选至少70％、最优选至少80％序列同一性的所述序列为所述FAS的直系同源物或功能变体，所述FAS包含或由如SEQ ID NO:5、6、15或16中所阐明的序列组成。
37.根据陈述1至8或27至36或43至56中任一项的重组原核或真核细胞或重组多核酸，其中所述FAS(亚基)包含或由一个或多个，优选所有以下结构域组成，优选从N-至C-末端为：乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰/棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)。
38.根据陈述1至8或27至37或43至56中任一项的重组原核或真核细胞或重组多核酸，包含多核酸序列，其中所述多核酸包含或由序列组成，所述序列优选从5’至3’编码一个或多个，优选所有以下结构域：乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)。
39.根据陈述1至8或24至38或43至56中任一项的重组原核或真核细胞或重组多核酸，其中所述TE源于表2中列出的生物体，优选来自不动杆菌属(Acinetobacter sp.)(例如A.baylyi或鲍氏不动杆菌(A.baumannii))的TE。
40.根据陈述1至8或27至39或43至56中任一项的重组原核或真核细胞或重组多核酸，其中所述TE包含或由以下组成：如SEQ ID NO:7中所阐明的序列，或与SEQ ID NO:7具有至少50％、优选至少60％、更优选至少70％、最优选至少80％序列同一性的序列。
41.根据陈述40的重组原核或真核细胞或重组多核酸，其中与SEQ ID NO:7具有至少50％、优选至少60％、更优选至少70％、最优选至少80％序列同一性的所述序列为所述TE的直系同源物或功能变体，所述TE包含或由如SEQ ID NO:7中所阐明的序列组成。
42.根据陈述1至8或27至41或43至56中任一项的重组原核或真核细胞或重组多核酸，其中编码所述ACP的所述核苷酸序列不是最3’或5’核苷酸序列。
43.根据陈述42的重组原核或真核细胞或重组多核酸，其中所述ACP编码序列是被所述TE编码序列替换的ACP编码序列。
44.根据陈述4至8或27至43或43至56中任一项的重组原核或真核细胞或重组多核酸，其中所述两个ACP编码序列串联排列。
45.根据陈述1至8或27至44或43至56中任一项的重组原核或真核细胞或重组多核酸，其中所述FAS(亚基)基因编码一个或多个，优选所有以下结构域：乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)。
46.根据陈述1至8或27至44或43至56中任一项的重组原核或真核细胞或重组多核酸，其中所述天然存在的FAS(亚基)源自含油细胞，优选含油酵母或含油微藻。
47.从5’至3’分别编码MPT、ACP、TE和KS的重组原核或真核细胞或重组多核酸，其中所述TE与所述MPT、ACP和/或KS相比优选是异源的。
48.从5’至3’分别编码MPT、TE、ACP和KS的重组原核或真核细胞或重组多核酸，其中所述TE与所述MPT、ACP和/或KS相比优选是异源的。
49.从5’至3’分别包含或编码以下的重组原核或真核细胞或重组多核酸：
-AT、ER、DH、MPT、ACP、TE、KS、KR和PPT；
-AT、ER、DH、MPT、TE、ACP、KS、KR和PPT；
-AT、ER、DH、MPT、ACP、TE、KS和KR；
-AT、ER、DH、MPT、TE、ACP、KS和KR；
-AT、ER、DH、MPT、ACP、TE和KS；
-AT、ER、DH、MPT、TE、ACP和KS；
-AT、ER、DH、MPT、ACP和TE；
-AT、ER、DH、MPT、TE和ACP；
-ER、DH、MPT、ACP、TE、KS、KR和PPT；
-ER、DH、MPT、TE、ACP、KS、KR和PPT；
-DH、MPT、ACP、TE、KS、KR和PPT；
-DH、MPT、TE、ACP、KS、KR和PPT；
-MPT、ACP、TE、KS、KR和PPT；
-MPT、TE、ACP、KS、KR和PPT；
-ACP、TE、KS、KR和PPT；
-TE、ACP、KS、KR和PPT；
-TE和ACP；
-ACP和TE；
-MPT、ACP、TE和KS；或
-MPT、TE、ACP和KS，
其中所述TE与所述AT、ER、DH、MPT、ACP、KS、KR和/或PPT相比优选是异源的。
50.重组原核或真核细胞或重组多核酸，包含根据陈述47至49中任一项的多核酸。
51.根据陈述47至50中任一项的重组原核或真核细胞或多核酸，从5’至3’分别编码AT、ER、DH、MPT、TE、ACP、KS、KR和PPT。
52.根据陈述47至51中任一项的重组原核或真核细胞或多核酸，从5’至3’分别编码DH、MPT、TE、ACP、KS、KR和PPT。
53.根据陈述47至52中任一项的重组原核或真核细胞或多核酸，其中所述AT、ER、DH、MPT、ACP、KS、KR和PPT分别衍生自天然存在的FAS(亚基)基因，优选I型FAS(亚基)基因。
54.根据陈述53的重组原核或真核细胞或多核酸，其中所述FAS(亚基)基因是真菌的，特别是酵母、原生生物、黏菌或藻类，特别是微藻FAS(亚基)基因。
55.根据陈述53至54中任一项的重组原核或真核细胞或多核酸，其中所述天然存在的FAS(亚基)基因来自红冬孢酵母属，优选圆红冬孢酵母或Aplanochytrium spp.，优选A.kerguelense。
56.根据陈述47至55中任一项的重组原核或真核细胞或多核酸，其中所述TE为酰基-CoA/ACP TE，优选短链或中链酰基-CoA/ACP TE。
57.由根据陈述47至56中任一项的重组多核酸编码的重组多肽。
58.包含根据陈述47至56中任一项的重组多核酸或编码根据陈述57的重组多肽的多核酸的重组载体。
59.根据陈述58的重组载体，其中所述载体为表达载体或重组载体。
60.重组原核或真核细胞，所述细胞包含根据陈述47至56中任一项的重组多核酸、根据陈述57的多肽、编码根据陈述57的多肽的多核酸或根据陈述58或59的载体。
61.根据先前陈述中任一项的重组原核或真核细胞，其中编码所述FAS、FAS1或FAS2和/或所述TE的多核酸是基因组整合的。
62.根据先前陈述中任一项的重组原核或真核细胞，其中所述TE编码核苷酸序列为异源TE编码核苷酸序列。
63根据先前陈述中任一项的重组原核或真核细胞，其为细菌细胞、真菌细胞或藻类细胞。
64根据先前陈述中任一项的重组原核或真核细胞，其为圆红冬孢酵母或Aplanochytrium kerguelense细胞。
65.根据先前陈述中任一项的重组原核或真核细胞，包含一个或多个编码AT、ER、DH、MPT、TE、ACP、KS、KR和PPT的多核酸。
66.根据先前陈述中任一项的重组原核或真核细胞，其为含油细胞，优选含油酵母或含油微藻细胞。
67.根据先前陈述中任一项的重组原核或真核细胞，其为单细胞生物体。
68.根据前面陈述中任一项的重组多核酸，其中在所述FAS(亚基)中AT、ER、DH、MPT、KS、KR和PPT的一个或多个是经修饰的。
在描述本发明的方法之前，应当理解的是，本发明不限于所描述的特定方法、组分、产物或组合，因为此类方法、组分、产物或组合当然可以变化。还应当理解的是，本文所使用的术语不旨在限制，因为本发明的范围将仅受所附权利要求的限制。
本发明的优选陈述(特征)和实施方案在下文中设定。除非明确地表明相反，否则如此定义的本发明的每个陈述和实施方案可以与任何其他陈述和/或实施方案组合。特别地，表明为优选或有利的任何特征可以与表明为优选或有利的任何其他特征或多个特征或陈述组合。至此，特别地通过一个或多个以上编号方面和实施方案1至68的任何一个或与任何其他陈述和/或实施方案的任何组合来体现本发明。将结合特定实施方案描述本发明，但是本发明不限于此，而是仅由权利要求限制。不应将权利要求中的任何参考标记诠释为限制范围。
当在本说明书和权利要求中使用术语“包含”时，不排除其他要素或步骤。本文使用的术语“包含”与“包括”或“含有”、同义，并是包括性的或开放式的，且不排除另外的未列举的成员、要素或方法步骤。应当领会的是，本文使用的术语“包含”包括术语“由……组成”以及术语“基本上由……组成”。
如本文所用，除非上下文另有明确规定，单数形式“一个”、“一种”和“该”包括单数和复数指示物。
由端点列举的数值范围包括归入在各自范围内的所有数字和分数，以及所列举的端点。
当提及可测量的数值例如参数、数量、时距等时，本文使用的术语“约”或“近似”是指涵盖指定值的+/-20％或更少、优选+/-10％或更少、更优选+/-5％或更少和仍更优选+/-1％或更少的变化，只要此类变化适合于在所公开的发明中实施。应当理解的是，修饰语“约”或“近似)”所指的数值本身也是具体并优选公开的。
鉴于术语“一个或多个”或“至少一个”，例如一组成员中的一个或多个或至少一个成员，本身是清楚的，通过进一步例证的方法，术语尤其涵盖提及任何一个所述成员，或任何两个或更多个所述成员，例如任何≥3、≥4、≥5、≥6或≥7等的所述成员，和多达所有所述成员。
本说明书中引用的所有参考文献都通过引用整体并入本文。特别地，本文具体提及的所有参考文献的教导通过引用并入本文。
除非另外定义，否则用于公开本发明的所有术语，包括技术和科学术语，具有本发明所属领域的普通技术人员通常理解的含义。通过进一步指导的方法，包括术语定义以更好地领会本发明的教导。
在整个说明书中对“一个实施方案”或“实施方案”的引用是指结合实施方案描述的特定特征、结构或特性包括在本发明的至少一个实施方案中。因此，整个说明书各个地方中出现的短语“在一个实施方案中”或“在实施方案中”不一定都，但是可以指的是相同的实施方案。此外，在一个或多个实施方案中，特定特征、结构或特性可以以任何适合的方式组合，如本领域技术人员从本公开中显而易见的。此外，虽然本文描述的一些实施方案包括其他实施方案中包括一些但不是其他特征，不同实施方案的特征的组合意为在本发明的范围内，如本领域技术人员应理解的。例如，在所附权利要求中，可以以任何组合使用任何要求保护的实施方案。
在以下本发明的详细描述中，参考了形成本文一部分的附图，并且其中仅通过例示的方式显示可以实践本发明的特定实施方案。应当理解的是，在不脱离本发明的范围的情况下，可以利用其他实施方案并且可以进行结构或逻辑上的改变。因此，以下详细描述不应被视为具有限制意义，并且本发明的范围由所附权利要求限定。
提供以下术语或定义仅用于帮助理解本发明。除非本文中具体定义，否则本文使用的所有术语具有对本发明领域技术人员而言相同的含义。从业者特别关注Sambrook etal.,Molecular Cloning:A Laboratory Manual,2nd ed.,Cold Spring Harbor Press,Plainsview,New York(1989)；和Ausubel et al.,Current Protocols in MolecularBiology(Supplement 47),John Wiley&Sons,New York(1999),Innis et al.,PCRProtocols:A Guide to Methods and Applications,Academic Press:San Diego,1990。对于本领域的定义和术语，微生物学的一般原理描述于例如Davis,B.D.et al.,Microbiology,3rd edition,Harper&Row,publishers,Philadelphia,Pa.(1980)。本文提供的定义不应诠释为具有比本领域普通技术人员所理解的更少的范围。
除非另有说明，否则可以以并且已经以本身已知的方式，作为对本领域技术人员清楚的方式实施所有未详细描述的方法、步骤、技术和操作。例如，再次参考了上文提及的标准手册、一般背景技术和其中引用的其他参考文献。
发明人惊奇地发现，尽管其结构复杂，但可以修饰微生物的脂肪酸合酶酶复合物以生成小至中链脂肪酸。
一方面，本发明涉及重组多核酸，所述重组多核酸包含脂肪酸合酶(FAS)编码序列或部分脂肪酸合酶(FAS)编码序列(即FAS亚基编码序列)和硫酯酶(TE)编码序列；任选地其中所述FAS(亚基)源自或衍生自天然存在的FAS(亚基)基因，所述天然存在的FAS(亚基)基因具有至少两个或恰好两个酰基载体蛋白(ACP)编码核苷酸序列，其中所述ACP编码核苷酸序列之一被失活、破坏，和/或由硫酯酶(TE)编码核苷酸序列替换。在相关方面，本发明涉及由所述多核酸编码的多肽、包含所述多核酸的载体或包含编码所述多肽的多核酸的载体，和包含所述多核酸、所述多肽或所述载体的宿主细胞。
一方面，本发明涉及重组多核酸，所述重组多核酸包含脂肪酸合酶(FAS)编码序列或部分脂肪酸合酶(FAS)编码序列(即FAS亚基编码序列)，源自或衍生自天然存在的FAS(亚基)基因，所述天然存在的FAS(亚基)基因具有酰基载体蛋白(ACP)编码核苷酸序列并缺乏硫酯酶结构域(TE)编码核苷酸序列，其中在酰基载体蛋白(ACP)编码核苷酸序列之前或之后添加异源硫酯酶结构域(TE)编码核苷酸序列。在相关方面，本发明涉及由所述多核酸编码的多肽、包含所述多核酸的载体或包含编码所述多肽的多核酸的载体，和包含所述多核酸、所述多肽或所述载体的宿主细胞。
本申请提供了所述重组多核酸在宿主细胞中的表达。因此，在相关方面，本发明涉及包含、表达或能够表达所述多核酸的重组宿主细胞，所述多核酸包含脂肪酸合酶(FAS)编码序列或部分脂肪酸合酶(FAS)编码序列(即FAS亚基编码序列)和硫酯酶(TE)编码序列；任选地其中所述FAS(亚基)源自或衍生自天然存在的FAS(亚基)基因，所述天然存在的FAS(亚基)基因具有至少两个或恰好两个酰基载体蛋白(ACP)编码核苷酸序列，其中所述ACP编码核苷酸序列之一被失活、破坏，和/或由硫酯酶(TE)编码核苷酸序列替换。在相关方面，本发明涉及由所述多核酸编码的多肽、包含所述多核酸的载体或包含编码所述多肽的多核酸的载体，和包含所述多核酸、所述多肽或所述载体的宿主细胞。
类似地，本发明涉及包含、表达或能够表达所述多核酸的重组宿主细胞，所述多核酸包含脂肪酸合酶(FAS)编码序列或部分脂肪酸合酶(FAS)编码序列(即FAS亚基编码序列)，源自或衍生自天然存在的FAS(亚基)基因，所述天然存在的FAS(亚基)基因具有酰基载体蛋白(ACP)编码核苷酸序列并缺乏硫酯酶结构域(TE)编码核苷酸序列，其中在酰基载体蛋白(ACP)编码核苷酸序列之前或之后添加异源硫酯酶结构域(TE)编码核苷酸序列。在相关方面，本发明涉及由所述多核酸编码的多肽、包含所述多核酸的载体或包含编码所述多肽的多核酸的载体，和包含所述多核酸、所述多肽或所述载体的宿主细胞。
应当理解的是，在本文所述的根据本发明的重组多核酸和重组宿主细胞中，当提及FAS时，意指功能性FAS，其可以包括FAS1、FAS2或FAS1和FAS2两者亚基。因此，重构功能性FAS所必需的所有单一酶结构域都存在于所述重组多核酸中(并且在所述细胞中得到表达或能够得到表达)。例如，重组多核酸可以包含并且宿主细胞可以包含或表达经工程化改造的FAS亚基，所述FAS亚基包含TE(例如FAS1或FAS2另外包含TE)和原生的或天然存在的或非经工程化改造的FAS亚基(例如FAS2或FAS1)以补充经工程化改造的FAS亚基并且以重建功能性FAS。
在某些实施方案中，本文所述的根据本发明的重组宿主细胞包含、表达或能够表达功能性天然的或天然存在的(任选地异源的)FAS，任选地包含FAS1和FAS2亚基。在此类实施方案中，(异源)TE编码序列整合在FAS编码序列中，如本文别处所述或单独提供。因此，FAS可以为另外包含TE的经工程化改造的原生的或天然存在的(异源的)FAS，或可以为补充有TE的原生的或天然存在的(异源的)FAS。
如本文所用，术语“核酸分子”、“多核苷酸”、“多核酸”、“核酸”可互换使用，并指任何长度的核苷酸的聚合形式，脱氧核糖核苷酸或核糖核苷酸两者中任一，或它们的类似物。多核苷酸可以具有任何三维结构，并可以行使任何功能，已知的或未知的。多核苷酸的非限制性实例包括基因、基因片段、外显子、内含子、信使RNA(mRNA)、转移RNA、核糖体RNA、核酶、cDNA、重组多核苷酸、分支多核苷酸、质粒、载体、任何序列的分离的DNA、控制区、任何序列的分离的RNA、核酸探针和引物。核酸可以是线性的或环状的。
本文使用的术语“重组”基本上是指非天然存在的或人工的(artificial)，即人造的(man-made)(多)核酸、(多)肽、细胞或生物体。该术语是指经工程化改造的或基因操作的(多)核酸、(多)肽、细胞或生物体。相反，本文使用的术语“天然存在的”是指非经操作的并可以在自然中找到的(多)核酸、(多)肽、细胞或生物体。如本文所用，术语“源自天然存在的FAS(亚基)”是指衍生自天然存在的FAS(亚基)多核酸或多肽的多核酸或多肽，其已经得到人工修饰(即重组多核酸或多肽)。应当理解的是，本文提及的FAS(亚基)基因、多核酸或蛋白质至少通过导入TE得到修饰，无论是否在多个ACP最初存在于所述FAS(亚基)的情况下通过替换ACP。然而，所述FAS(亚基)的进一步修饰也是可能的。举例说明，一个或多个所述FAS(亚基)的功能性结构域(例如AT、ER、DH、MPT、KS、KR和/或PPT)可以得到修饰，包括用例如但不限于直系同源FAS(亚基)的相应功能性结构域替换此类功能性结构域。
如本文所用，术语“多肽”、“蛋白质”、“肽”和“氨基酸序列”可互换使用，并指任何长度的氨基酸的聚合形式，其可以包括编码的和非编码的氨基酸、以化学方式或生物化学方式修饰的或衍生化的氨基酸和具有经修饰的肽骨架的多肽。如本文所用，氨基酸残基将以它们的全名或根据标准的三字母或单字母氨基酸代码表示。
本文通常使用的术语“基因”是指含有编码序列、启动子和任何其他宿主细胞中表达所需的调节区的核酸序列。
如本文所用，术语“启动子”是指位于转录起始位点上游50bp以内的非翻译序列并且其控制结构基因的转录起始。通常它位于结构基因的翻译起始密码子上游(即5’)的约1至1000bp，优选1-500bp，特别是1-100bp以内。类似地，术语“终止子”是指位于至结构基因的翻译终止密码子下游(即3’)的非翻译序列(通常在约1至1000bp，更典型1-500碱基对并且特别是1-100碱基对以内)并且其控制结构基因的转录终止。如果它的位置相对于编码序列的位置使得启动子或终止子，视情况而定，行使它的转录控制功能，则启动子或终止子与编码序列为“可操作地相连或连接”。
如本文所用，术语“异源”或“外源”是指所考虑的基因、核酸或蛋白质序列、结构域或编码序列源自关注或研究的宿主生物体外的事实，或者是指所考虑的基因或编码序列对宿主而言不是原生的或内源的，而是源自或已经从不同的细胞类型或与受体宿主不同的物种的生物体克隆。
术语“原生”或“内源”在本文中用于在宿主菌株的野生型细胞的基因组内发现的(除了不影响功能的个体对个体的变异)遗传物质(例如基因、启动子或终止子等)。
当提及宿主细胞中基因的表达时本文使用的术语“过表达”是指在所述宿主细胞中以高于天然的水平表达的事实。这可以暗指其为外源基因，不在宿主细胞中天然表达，或者内源基因已经以增加在所述宿主细胞中的表达的方式进行了修饰。
当提及存在于宿主细胞中的基因、基因序列或编码结构域的序列时本文使用的术语“失活”是指基因产物在所述宿主细胞中不表达或没有活性的事实。这通常暗指内源基因、基因序列或编码结构域的序列已经以不再允许在所述宿主细胞中表达的方式进行了修饰，但也可以暗指基因已经以确保基因产物不再具有活性的方式进行了修饰，并且包括基因、基因序列或编码结构域的序列的部分或完整，优选完整的缺失。
“编码“是指核酸序列或它的部分，由于所讨论的生物体的遗传密码，对应于特定氨基酸序列，例如所需多肽或蛋白质的氨基酸序列。举例说明，“编码”特定多肽或蛋白质的核酸可以涵盖基因组的，hnRNA、pre-mRNA、mRNA、cDNA，重组或合成核酸。
优选地，编码特定多肽或蛋白质的核酸可以包含编码所述多肽或蛋白质的开放阅读框(ORF)。“开放阅读框”或“ORF”是指一连串编码核苷酸三联体(密码子)，其以翻译起始密码子开始并以本身已知的翻译终止密码子结束，并不含有任何内部框内翻译终止密码子，并潜在地能够编码多肽。因此，该术语可以与本领域中使用的“编码序列”同义。在某些实施方案中，本文所述的多核酸包含含有ACP编码序列和TE编码序列(优选彼此相邻)的开放阅读框。
本文使用的术语“脂肪酸”是指一大群有机酸的任意项，特别是在动植物油脂中发现的那些。特征性地由具有偶数碳原子的饱和或不饱和脂肪族化合物组成，该组酸包括棕榈酸、硬脂酸和油酸，但还包括短链脂肪酸(SCFA)和中链脂肪酸(MCFA)。短链脂肪酸(SCFA)是具有少于六个碳的脂肪族尾部的脂肪酸(例如丁酸)，而中链脂肪酸(MCFA)是具有通常6-12个碳的脂肪族尾部的脂肪酸。
本文使用的术语“饱和脂肪酸”是指其中所有碳原子之间的键都是单键的任何脂肪酸。
本文使用的术语“不饱和脂肪酸”是指其中至少一个碳原子之间的键是双键的任何脂肪酸。
术语“脂肪酸合酶(fatty acid synthase)”、“脂肪酸合成酶(fatty acidsynthetase)”或“FAS”是指催化脂肪酸合成的酶复合物或多酶蛋白质。FAS不是单一酶，而是由不同酶或结构单元或结构域组成的多模式酶系统。FAS有两个主要种类。I型FAS利用单个或两个大的多功能多肽并对哺乳动物和真菌两者都是常见的(尽管真菌和哺乳动物合酶的结构排列不同)。在CMN细菌群体(棒状杆菌(corynebacteria)、分枝杆菌(mycobacteria)和诺卡氏菌(nocardia))中也发现了I型脂肪酸合酶系统。在古菌和细菌中发现了II型FAS，并且其特征在于使用分立的单功能酶进行脂肪酸合成。I型FAS和II型FAS延伸和还原的机制相同，因为II型FAS酶的结构域与I型FAS多酶多肽中的结构域对应物在很大程度上同源。FAS基因通常编码多个酶单位或结构域，例如乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)、硫酯酶(TE)、丙二酰-乙酰转移酶(MAT)，其中一些或全部通常构成功能性FAS复合物。举例说明并不限于，真菌FAS通常包含以下酶结构域：乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)。动物，特别是哺乳动物FAS通常包括以下酶结构域：酮脂酰合酶(KS)、丙二酰-乙酰转移酶(MAT)、脱水酶(DH)、烯酰还原酶(ER)、酮脂酰还原酶(KR)、酰基载体蛋白(ACP)、硫酯酶(TE)。I型FAS由一个或两个分立的基因或基因序列编码。在FAS由两个基因或基因序列编码的情况下，单一基因(序列)编码FAS1(也称为FAS亚基β或FASβ链)和FAS2(也称为FAS亚基α或FASα链)。哪个亚基或链中包含哪个酶结构域是可变的。举例说明并不限于，酿酒酵母FAS包含(从N-至C-末端排列)FAS2(α链)上的ACP、KS、KR和PPT并包含(从N-至C-末端排列)FAS1(β链)上的AT、ER、DH和MPT。新型隐球酵母(Cryptococcus neoformans)FAS包含(从N-至C-末端排列)FAS2(α链)上的KS、KR和PPT并包含(从N-至C-末端排列)FAS1(β链)上的AT、ER、DH、MPT和ACP。圆红冬孢酵母FAS包含(从N-至C-末端排列)FAS2(α链)上的DH、MPT、ACP、KS、KR和PPT并包含(从N-至C-末端排列)FAS1(β链)上的AT和ER。应当注意的是，圆红冬孢酵母包含串联排列的两个ACP结构域。与上述相反，其他生物体将FAS作为单一基因编码，因此FAS是单一多肽。举例说明并不限于，玉米黑粉菌(Ustilago maydis)和Aplanochytrium kerguelense FAS包含在单一多肽(从N-至C-末端排列)上的乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)。应当注意的是，Aplanochytrium kerguelense包含串联排列的两个ACP结构域。如本领域技术人员所领会的，ACP的“串联”排列是指在位于蛋白质中彼此相邻的(即相邻的(adjacent)，如本文他处所定义的，通过接头可选择地连接)两个(或更多个)ACP结构域，没有居间的其他FAS结构域，即没有居间的AT、ER、DH、MPT、KS、KR、MAT、TE或PPT。如本文所用，术语“脂肪酸合酶亚基”、“脂肪酸合成酶亚基”或“FAS亚基”是指其中不存在全功能性FAS所有结构域的部分FAS。举例说明，FAS亚基可以缺乏乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)、丙二酰-乙酰转移酶(MAT)的一个或多个。进一步举例说明，应当理解的是，FAS亚基虽然缺乏一个或多个酶结构域，但存在的结构域优选是完整的和功能性的，即这些结构域优选不被截短。进一步举例说明，本文提及的FAS亚基可以是FAS1或FAS2，如本文他处所述。优选地，本文提及的FAS亚基包含至少一个ACP结构域，例如一个ACP或两个ACP结构域，任选地串联的两个ACP结构域。例示性的FAS亚基可以例如衍生自图1。应当理解的是，本文提及的FAS亚基不含有完整功能所必需的所有酶结构域。为了实现完整功能，可以需要用FAS亚基中缺失的酶结构域补充本文提及的FAS亚结构域。可以有利地作为单一多肽或作为编码单一多肽的单一ORF提供此类缺失的结构域。
对于动物和真菌源的代表性I型FAS的结构域配置，参考Maier et al.(2008)“Thecrystal structure of a mammalian fatty acid synthase”；Science；321(5894):1315-22(doi:10.1126/science.1161269)；和Lomakin et al(2007)“The crystal structureof yeast fatty acid synthase,a cellular machine with eight active sitesworking together”；Cell；129(2):319-32，其全部内容通过引用并入本文。
在优选实施方案中，本文所述的FAS为I型FAS，包括FAS1和/或FAS2。因此，在某些实施方案中，本文所述的FAS是或包含FAS1或FAS2。优选地，本文所述的FAS是或包含FAS1或FAS2，其中所述FAS1或FAS2包含ACP结构域。在某些实施方案中，当提及插入在ACP上游或下游的FAS中的TE结构域时，在FAS由两个分立的亚基FAS1和FAS2组成的情况下，将TE结构域插入在含有ACP的FAS亚基上。因此，在某些实施方案中，本文提及的FAS可以与FAS1或FAS2有关，无论哪个包含ACP。
在某些实施方案中，在FAS(亚基)，优选含有ACP的FAS亚基中提供或插入如本文提及的TE，例如优选异源TE。在某些实施方案中，在FAS(亚基)中不提供或插入如本文提及的TE，例如优选异源TE。在某些实施方案中，在所述FAS(亚基)的ACP上游或下游的FAS(亚基)中，即与所述FAS(亚基)的ACP相邻或直接相邻，优选所述FAS(亚基)的ACP的直接上游或下游，提供或插入如本文提及的TE，例如优选异源TE，即TE核酸序列直接地ACP的核酸序列的5’或3’，或TE多肽序列直接地ACP的多肽序列的N-末端或C-末端。如在此上下文中所用，术语“直接地”表明在ACP和TE序列之间不存在另外的结构域，特别是不存在另外的酶结构域(例如AT、ER、DH、MPT、KS、KR或PPT)，特别是不存在另外的功能性酶结构域。技术人员将理解，如本领域已知的，可能地在ACP和TE序列之间可以存在接头序列。然而，也可以不存在接头序列。应当理解的是，接头序列不是并不包含(功能性)酶结构域。在某些实施方案中，存在至多200核苷酸的接头序列，例如至多100核苷酸。在某些实施方案中，存在至多70核苷酸的接头序列，例如至多35核苷酸。在某些实施方案中，由SEQ ID NO:110的核酸序列，或它的功能性片段或变体编码接头序列。在某些实施方案中，接头具有SEQ ID NO:111的氨基酸序列，或它的功能性片段或变体，或编码SEQ ID NO:111的氨基酸序列的核酸序列。
在优选实施方案中，本文所述的FAS(亚基)是源自单细胞生物体的FAS(亚基)，所述单细胞生物体优选选自包含或由酵母、原生生物、黏菌和微藻，优选酵母或微藻组成的组。
在优选实施方案中，本文所述的FAS(亚基)是源自含油生物体，优选含油单细胞生物体的FAS(亚基)。
在优选实施方案中，本文所述的FAS是源自含油单细胞生物体的I型FAS，所述含油单细胞生物体优选选自包含或由酵母、原生生物、黏菌和微藻，优选酵母或微藻组成的组。
在某些实施方案中，FAS(亚基)可以是人工的，因为不同的酶结构域可以源自不同的生物体。在某些实施方案中，不同的酶结构域可以源自相同的生物体。在某些实施方案中，不同的FAS亚基可以源自不同的生物体。在某些实施方案中，不同的FAS亚基可以源自相同的生物体。
在某些实施方案中，本文使用的FAS，优选I型FAS是指包含ACP和一个或多个，优选全部其他FAS酶结构域(或编码相应结构域的多核酸)的单一多肽。在某些实施方案中，本文使用的FAS是指包含ACP和一个或多个，优选全部其他FAS1和/或FAS2结构域(或编码相应结构域的多核酸)的FAS1和/或FAS2。在某些实施方案中，由单一开放阅读框编码本文所提及的TE和ACP(其可以存在于作为单一蛋白质组成的FAS或由FAS1和FAS2亚蛋白质组成的FAS中)。因此，一方面，本发明涉及编码真菌脂肪酸合酶(FAS)亚基(例如FAS1或FAS2)的重组多核酸，所述亚基(例如FAS1或FAS2)包含ACP和硫酯酶(TE)，优选异源TE。
技术人员将理解如果FAS由FAS1和FAS2组成，可以在这些之一中插入TE，优选相邻的并且在其中含有ACP。在生产脂肪酸(或它们的衍生物)的方法中，如本文他处提及的，在某些实施方案中，如果此类宿主生物体天然含有FAS(其可以是或可以不是与经工程化改造的FAS衍生自的生物体相同的生物体)，则只可以在宿主生物体中导入如此经工程化改造的FAS1(或FAS2)。以这种方式，宿主生物体可以补充其他FAS亚基(例如，在内源性表达FAS2的宿主生物体中导入含有FAS1的TE)。
如本文所提及的，本文所提及的(天然存在的)FAS(亚基)可以在某些实施方案中包含至少两个ACP结构域，即FAS(亚基)基因编码至少两个ACP结构域。在本文所述的重组多核酸或多肽序列、载体、细胞或方法中，可以用TE结构域或TE编码序列替换两个ACP结构域或ACP编码序列之一。技术人员将理解如果在天然存在的FAS(亚基)中存在超过两个ACP结构域，可以用TE结构域替换超过一个ACP结构域，只要至少一个ACP结构域仍然存在。在某些实施方案中，其中FAS由不同的FAS亚基组成，优选仅一个亚基(或编码亚基的核苷酸序列)包含ACP(或多个ACP)，而另一个亚基(或编码亚基的核苷酸序列)不包含ACP。
如本文所提及的，本文所提及的(天然存在的)FAS(亚基)优选包含至少一个ACP结构域，即FAS(亚基)基因编码至少一个ACP结构域并缺乏TE结构域，即FAS(亚基)基因不编码TE结构域。在本文所述的重组多核酸或多肽序列、载体、细胞或方法中，优选在ACP结构域之前或之后添加TE结构域，或已经在ACP编码序列之前或之后添加了TE编码序列。
表1中列出了用于本文所述的根据本发明的方面中的适合的(天然存在的)FAS的非限制性实例，所述FAS具有至少两个ACP结构域。因此，在某些实施方案中，(天然存在的)FAS如表1中所说明或衍生自表1中列出的生物体。通过扩展，各个FAS结构域(或它们的组合)，例如乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)、硫酯酶(TE)、丙二酰-乙酰转移酶(MAT)，可以衍生自表1中列出的(天然存在的)FAS。
表1：来自适合于根据本发明的实施方案使用的生物体的FAS
技术人员将理解除了表1中列出的FAS/生物体，可以鉴定根据本发明适合的另外的FAS和生物体，例如不限于也包含至少两个ACP结构域的FAS(亚基)。这可以例如通过序列比对来完成。用于比较序列和确定序列同一性的方法是本领域公知的。举例说明，序列同一性的百分比是指在比对这些序列后两个序列之间相同的核酸或氨基酸的百分比。可以用本领域已知的各种不同的程序和算法实施和计算同一性的比对和百分比。优选的比对算法包括BLAST(Altschul,1990；例如可在NCBI网站获得)和Clustal(综述于Chenna,2003；例如可在EBI网站获得)。优选地，BLAST用于计算两个序列之间同一性的百分比，例如由Tatusova和Madden 1999(FEMS Microbiol Lett 174:247-250)描述的“Blast 2序列”算法，例如使用已发布的默认设置或其他适合的设置(例如，对于BLASTN算法：打开缺口代价＝5,延伸缺口代价＝2,错配罚分＝-2,匹配得分＝1,缺口x_dropoff＝50,期望值＝10.0,字长＝28；或对于BLASTP算法：矩阵＝Blosum62,打开缺口代价＝11,延伸缺口代价＝1,期望值＝10.0,字长＝3)。
举例说明，可以例如将ACP序列(以及一般FAS(亚基)序列，或FAS(亚基)的选择的结构域序列，例如乙酰转移酶(AT)、烯酰还原酶(ER)、脱水酶(DH)、丙二酰-棕榈酰转移酶(MPT)、酰基载体蛋白(ACP)、酮脂酰合酶(KS)、酮脂酰还原酶(KR)、磷酸泛酰巯基乙胺基转移酶(PPT)、硫酯酶(TE)、丙二酰-乙酰转移酶(MAT))与Zhu et al.(2012)“A multi-omicmap of the lipid-producing yeast Rhodosporidium toruloides”；Nat Commun；3:1112(doi:10.1038/ncomms2112)中描述的序列比对，通过引用将其全部明确地并入本文。
在某些实施方案中，(天然存在的)FAS(亚基)具有如SEQ ID NO:8、9、10、14或20中所阐明的蛋白质序列，或它们的功能性片段、变体或直系同源物，或者具有编码如SEQ IDNO:8、9、10、14或20中所阐明的蛋白质序列的编码序列，或它们的功能性片段、变体或直系同源物。
在某些实施方案中，作为通过用TE结构域替换ACP结构域修饰的FAS(亚基)具有如SEQ ID NO:11或12中所阐明的蛋白质序列，或它们的功能性片段、变体或直系同源物，或者具有编码如SEQ ID NO:11或12中所阐明的蛋白质序列的编码序列，或它们的功能性片段、变体或直系同源物。
在某些实施方案中，FAS(亚基)通过在ACP结构域之前或之后添加TE结构域得到修饰，并具有如SEQ ID NO:17或18中所阐明的蛋白质序列，或它们的功能性片段、变体或直系同源物，或者具有编码如SEQ ID NO:17或18中所阐明的蛋白质序列的编码序列，或它们的功能性片段、变体或直系同源物。
在某些实施方案中，FAS(亚基)或编码多核苷酸序列的FAS(亚基)，作为通过导入TE结构域(或用TE结构域替换ACP结构域)得到修饰，包含或由以下组成(多核苷酸序列编码，优选从5’至3’)：
-AT、ER、DH、MPT、ACP、TE、KS、KR和PPT；
-AT、ER、DH、MPT、TE、ACP、KS、KR和PPT；
-AT、ER、DH、MPT、ACP、TE、KS和KR；
-AT、ER、DH、MPT、TE、ACP、KS和KR；
-AT、ER、DH、MPT、ACP、TE和KS；
-AT、ER、DH、MPT、TE、ACP和KS；
-AT、ER、DH、MPT、ACP和TE；
-AT、ER、DH、MPT、TE和ACP；
-ER、DH、MPT、ACP、TE、KS、KR和PPT；
-ER、DH、MPT、TE、ACP、KS、KR和PPT；
-DH、MPT、ACP、TE、KS、KR和PPT；
-DH、MPT、TE、ACP、KS、KR和PPT；
-MPT、ACP、TE、KS、KR和PPT；
-MPT、TE、ACP、KS、KR和PPT；
-ACP、TE、KS、KR和PPT；
-TE、ACP、KS、KR和PPT；
-TE和ACP；
-ACP和TE；
-MPT、ACP、TE和KS；或
-MPT、TE、ACP和KS；
其中所述TE优选与所述AT、ER、DH、MPT、ACP、KS、KR和/或PPT相比是异源的，并且其中所述AT、ER、DH、MPT、ACP、KS、KR和/或PPT优选彼此相比不是异源的。
如本文所述，在某些实施方案中，用TE结构域替换天然存在的FAS(亚基)中的(至少)一个ACP结构域，所述FAS(亚基)包含(至少)两个ACP结构域。在某些实施方案中，在FAS(亚基)的ACP的上游或下游(直接地)导入TE。
如本文所用，术语“替换”是指删除ACP结构域或ACP编码核苷酸序列并在缺失的ACP结构域或ACP编码核苷酸序列的位置插入TE结构域或TE编码核苷酸序列。可以通过本领域已知的技术，包括标准克隆技术完成用TE结构域或TE编码序列替换ACP结构域或ACP编码核苷酸序列。举例说明并不限于，可以在体外克隆FAS基因序列或部分FAS基因序列(即FAS亚基，例如包含编码ACP结构域的核苷酸序列的部分FAS核苷酸序列；或例如包含编码ACP结构域的核苷酸序列的(部分)FAS1或FAS2序列)。通过标准克隆技术，包括例如使用PCR、限制性内切酶或体外同源重组，可以去除并用编码TE的核苷酸序列替换编码ACP的核苷酸序列。在某些实施方案中，可以通过本领域已知的技术在本文他处定义的原核或真核细胞中导入所得的重组多核酸。技术人员将理解，用TE编码序列替换ACP编码序列优选但不是必需，需要删除整个ACP编码序列。它可以是初始ACP序列的部分或全部仍然存在。例如，可以将TE编码序列插入到ACP编码序列中，例如将ACP编码序列扰乱到不产生功能性ACP的程度。然而，优选地，删除ACP的整个编码序列或ACP的大部分编码序列，例如删除至少80％，优选至少90％，更优选至少95％的ACP的编码序列。
如本文所用，术语“硫酯酶”是指催化酯水解成酸和醇(特别是在硫醇基团上)的酶。硫酯酶的非限制性实例包括不限于乙酰-CoA水解酶、棕榈酰-CoA水解酶、琥珀酰-CoA水解酶、甲酰-CoAA水解酶、酰基-CoAA水解酶等。本文提及的优选硫酯酶为酰基-ACP/CoAA硫酯酶，优选对SCFA和/或MCFA具有特异性并产生或导致其释放的酰基-ACP/CoA A硫酯酶。表2中列出了用于本文所述的根据本发明的方面的适合的硫酯酶的非限制性实例。因此，在某些实施方案中，硫酯酶如表2中所说明或衍生自表2中列出的生物体。
表2：来自适合于根据本发明的实施方案的生物体的TE
技术人员将理解，除了表2中列出的TE/生物体，可以鉴定也包含用于根据本发明的使用适合的TE的另外的TE和生物体。这可以例如通过序列比对来完成，如本文他处所详述。
在某些实施方案中，TE选自以下：AcTesA(衍生自Acinetobacter baylyi；以genbank登录号WP_004921669.1表示)、TEII(衍生自小家鼠；以genbank登录号NP_666033.1表示)、TEII(衍生自褐家鼠；以genbank登录号NM_022705.1表示)、YpTesB(衍生自鼠疫耶尔森氏菌；以genbank登录号CAL21736.1表示)、fatB1(衍生自椰子；以genbank登录号AEM72519.1表示)。
在某些实施方案中，TE具有如SEQ ID NO:13中所阐明的蛋白质序列，或它的功能性片段、变体或直系同源物，或者具有编码如SEQ ID NO:13中所阐明的蛋白质序列，或它的功能性片段、变体或直系同源物的编码序列。
一方面，本发明涉及由本文所述的根据各种实施方案的多核酸编码的蛋白质或多肽。
另一方面，本发明涉及包含如本文所述的根据各种实施方案的多核酸的载体，或包含编码如本文所述的根据各种实施方案的多肽的多核酸的载体。
“载体”是指多核酸分子，优选DNA分子，其衍生自，例如质粒、噬菌体或植物/动物病毒，其中可以插入或克隆多核苷酸。载体优选含有一个或多个唯一限制酶切位点并可以能够在定义的宿主细胞中自主复制，或可与所定义宿主的基因组整合，使得克隆的序列是可复制的。载体的选择通常取决于载体与要导入载体的宿主细胞的相容性。
根据本发明的实施方案，表达构建体为表达载体，适合于转化到宿主生物体，优选细菌中，并适合于在经转化的宿主细胞中的如本文所述的根据本发明的多核酸的维持和/或表达。
本发明的载体在所选择的宿主菌株中作为克隆载体或表达载体可以是可操作的。本领域技术人员已知许多载体，并且选择适当的载体是一个选择问题。载体可以例如是pASK-IBA3C表达载体(IBA-life sciences)、pUR5750转化载体(de Groot etal.1998Nature Biotechnology 16,839–842)、pCGHT3转化载体(Chambers etal.1988Gene,Volume 68,Issue 1:15；Scholtmeyer etal.2001Appl.Environ.Microbiol.67(1):481)。
“表达载体”是可以用于转化所选择的宿主细胞并在所选择的宿主细胞中提供编码序列的表达的构建体。表达载体可以例如是克隆载体、双元载体或整合(例如重组载体，包括同源重组，或随机整合)载体。本发明因此也涉及包含本文所述的任何多核酸的载体。所述载体可以进一步包含用于在所述宿主细胞中控制多核酸表达的调节序列。通常，表达牵涉使用能够在宿主细胞中有效复制的表达载体，使得宿主细胞累积表达载体的许多拷贝，并且反过来合成由表达载体编码的高水平的所需产物。
本文使用的术语“调节序列”和“控制序列”在广泛的上下文中提及，并且是指能够驱动和/或调节与它们连接的(共价连接)和/或可操作地连接的序列表达的调节核酸序列。控制序列根据预想的宿主生物体和待表达序列的性质而不同。对于原核生物中蛋白质的表达，控制序列通常包括启动子、核糖体结合位点和终止子。在真核生物中，控制序列通常包括启动子、终止子以及在某些情况下，增强子和/或5’和3’非翻译序列。术语“控制序列”意指包括，最低限度，表达必需的所有组分，并还可以包括另外的有利组分。根据本发明的优选实施方案，控制序列在如本文他处所定义的宿主细胞中是可操作的。术语“控制序列”涵盖能够在宿主细胞中激活或增强核酸分子表达的启动子或序列。
表达和克隆载体通常含有由宿主生物体识别并与编码感兴趣的多肽的核酸(共价并且)可操作地连接的启动子。启动子是位于结构基因起始密码子上游(5')的非翻译序列(通常在约100至1000bp以内)，其控制与它们可操作地连接的特定核酸序列，例如编码本文定义的融合蛋白的序列的转录和翻译。此类启动子通常分为两类，诱导型和组成型。诱导型启动子是响应于培养条件的一些变化，例如营养物的存在或不存在或温度的变化，在其控制下从核酸启动增加的转录水平的启动子。此时，公知由各种潜在宿主细胞识别的大量启动子。通过限制酶消化从源核酸中去除启动子并将分离的启动子序列插入到载体中来将这些启动子与编码感兴趣的多肽的核酸可操作地连接。天然存在的启动子序列和许多异源启动子两者可以用于直接扩增和/或表达感兴趣的多肽。通常，含有衍生自与宿主细胞相容的物种的启动子和控制序列的质粒载体与这些宿主一起使用。载体通常携带一个或多个能够在经转化的细胞中提供表型选择的复制位点以及标记序列。
根据本发明的一个实施方案，载体包含组成型启动子。适合于根据本发明的构建体和方法的组成型启动子的实例包括但不限于CaMV35S启动子、GOS2、肌动蛋白启动子、泛素启动子、硫解酶启动子。
根据本发明的另一个实施方案，载体包含诱导型启动子。适合于根据本发明的构建体和方法的诱导型启动子的实例包括但不限于lac启动子或木糖诱导型启动子。
任选地，本发明的表达载体还含有对于终止转录和稳定mRNA所必需的序列，并因此可以含有一个或多个转录终止序列。术语“转录终止序列”涵盖转录单元末端的控制序列，其表示转录的3’加工和终止。可以在表达构建体中掺入另外的调节元件，例如转录或翻译增强子。
本发明的表达构建体可以进一步包括对特定细胞类型中的维持和/或复制所需的复制起点。一个实例是当需要将表达构建体在细胞中作为附加型遗传因子(例如质粒或粘粒分子)维持时。优选的复制起点包括但不限于f1-ori、colE1 ori和Gram+细菌复制起点。
表达构建体可以任选地包含选择标记基因。如本文所用，术语“选择标记基因”包括赋予细胞表型的任何基因，在所述细胞中表达它以便于鉴定和/或选择用本发明的表达构建体转染或转化的细胞。典型的选择标记基因编码以下蛋白质：(a)赋予对抗生素或其它毒素，例如氯毒素、吉欧霉素(来自印度异壁链霉菌(Streptoalloteichus hindustanus)的sh ble基因)、genetecin、蜜二糖酶(MEL5)、潮霉素(来自大肠杆菌的氨基糖苷类抗生素耐药性基因)、氨苄青霉素、四环素或卡那霉素(Tn903卡那霉素抗性基因)的抗性，(b)补充细胞的营养缺陷型缺陷。营养缺陷型缺陷的两个突出实例为氨基酸亮氨酸缺陷(例如LEU2基因)或尿嘧啶缺陷(例如URA3基因)。乳清苷-5’-磷酸脱羧酶阴性(ura3-)的细胞不可以在缺乏尿嘧啶的培养基上生长。因此可以使用功能性URA3基因作为具有尿嘧啶缺陷的细胞上的标记，并且可以在缺乏尿嘧啶的培养基上选择成功的转化体。只有用功能性URA3基因转化的细胞能够合成尿嘧啶并在此类培养基上生长。如果野生型菌株不具有尿嘧啶缺乏(例如，和东方伊萨酵母的情况一样)，必须制备具有缺陷的营养缺陷型突变体，以便使用URA3作为菌株的选择标记。本领域公知完成此目的的方法。优选的选择标记包括吉欧霉素抗性基因、G418抗性基因、潮霉素抗性基因。选择标记组件通常还包括与选择标记基因可操作地连接的启动子和终止子序列，并且其在宿主菌株中是可操作的。也可以使用视觉标记基因，包括例如beta-葡糖醛酸糖苷酶(GUS)、荧光素酶和绿色荧光蛋白(GFP)。
含有一个或多个上文列出的组分并包括所需的编码和控制序列的适合的载体的构建采用标准连接技术。将分离的质粒或核酸片段切割、剪裁，并以生成所需质粒的所需形式重新连接。
作为上述表达载体的替代，无论是诱导型或组成型表达载体，在某些实施方案中，如本文所述的载体不包含负责表达由本文所述的多核酸编码的蛋白质的调节序列。例如，这可以是整合或重组载体的情况，如本领域中已知的。举例说明并不限于，通过重组，可以用如本文所述的根据本发明的多核酸替换内源FAS基因或它的一部分。以这种方式，在内源启动子的控制下，初始包含天然存在的FAS基因(例如但不限于编码(至少)两个ACP结构域的FAS基因)的宿主细胞现在包含含有TE结构域的重组FAS基因。或者，可以实施随机整合，在这种情况下，如本文所述的根据本发明的多核酸优选与一个或多个调节序列(如本文他处所述，例如组成型或诱导型启动子)可操作地连接。通过设计和构建适当的载体并用那些载体转化宿主菌株，在一个或多个步骤中完成宿主菌株的基因修饰。可以使用如本领域已知的电穿孔和/或化学(例如基于氯化钙或乙酸锂的)转化方法或根癌农杆菌(Agrobacterium tumefaciens)介导的转化方法。载体可以用特定的限制酶切割或用作环状DNA。用于宿主菌株基因修饰的载体可以是任何载体，只要它可以整合在宿主菌株的基因组中。
可以通过利用由标记基因贡献的属性或通过由插入基因贡献的其他特征(例如能够生产脂肪酸，不能生产乳酸或乳酸盐，不能生产乙酸或乙酸盐，或能够在特定底物上生长)以已知方式选择成功的转化体(包含如文本所述的附加或基因组整合的多核酸或载体)。可以通过PCR或Southern分析实施筛选，以证实发生了所需的插入和缺失，证实拷贝数并确认基因整合到宿主菌株基因组中的点。可以使用已知测定方法证实由插入基因编码的酶的活性和/或由缺失基因编码的酶的活性的缺乏。
一方面，本发明涉及宿主细胞，特别是重组宿主细胞，优选原核或真核宿主细胞，所述细胞表达或能够表达本文所述的FAS和TE，或包含本文所述的多核酸、多肽或载体。优选地，宿主细胞表达或能够表达(例如诱导地或条件性表达)由本文所述的根据本发明的多核酸编码的多肽。如上文所详述的，对重组宿主细胞的提及暗指宿主细胞包含至少一个外源或异源(即非天然)遗传因子，更具体地，编码FAS(亚基)的多核苷酸序列和/或编码TE的多核酸。更具体地，编码FAS(亚基)的多核酸是编码不包含TE结构域的FAS的多核酸，更具体地真菌FAS。在特定实施方案中，异源多核酸为天然存在的核酸。
在相关方面，本发明涉及宿主细胞，特别是重组宿主细胞，优选原核或真核宿主细胞，所述细胞表达或能够表达(例如诱导地或条件性表达)源自天然存在的FAS(亚基)基因(优选缺乏TE)的FAS(亚基)基因序列，并进一步表达或能够表达(异源)TE，例如插入(异源)TE核苷酸序列(优选在所述FAS(亚基)的ACP的(直接地)上游或下游)，其中任选地所述FAS(亚基)具有至少两个ACP编码核苷酸序列(优选串联)，其中用TE编码核苷酸序列替换所述ACP编码核苷酸序列之一，其中所述FAS(亚基)、ACP和TE如本文他处详述。技术人员将理解，当提及FAS亚基，例如包含一个或多个ACP和经工程化改造的TE(优选在ACP的直接地上游或下游)的FAS亚基(为了重建能够生产S/MCFA的功能性FAS)，可以需要提供单独的亚基，其单独的亚基包含FAS亚基缺乏的酶结构域，所述FAS亚基包含一个或多个ACP和经工程化改造的TE。因此，在某些实施方案中，本发明涉及本文提及的宿主细胞，表达或能够表达(例如诱导地或条件性表达)多个FAS亚基基因序列，例如两个FAS亚基基因序列，所述FAS亚基基因序列源自天然存在的FAS基因(优选缺乏TE)并插入(异源)TE核苷酸序列(优选在含有ACP的FAS亚基的ACP的(直接地)上游或下游)，其中任选地所述FAS亚基之一具有至少两个ACP编码核苷酸序列(优选串联)，其中用TE编码核苷酸序列替换所述ACP编码编码核苷酸序列之一，其中所述FAS(亚基)、ACP和TE如本文他处详述。在某些实施方案中，宿主细胞包含多个FAS亚基，使得生产S/MCFA，即在组合的多个FAS亚基上提供所有必需的酶结构域。
在某些实施方案中，宿主细胞为细菌细胞、真菌细胞(优选酵母细胞)或藻类细胞(优选微藻细胞)。
在优选实施方案中，宿主细胞为含油生物体或单细胞生物体，优选含油单细胞生物体。在特定实施方案中，宿主细胞为酵母细胞，例如含油酵母细胞。在另一个特定实施方案中，宿主选自以下：酵母属(Saccharomycetes sp)(例如酿酒酵母，例如酿酒酵母PWY12)、亚罗酵母(Yarrowia sp.)(例如解脂亚罗酵母(Y.lipolytica))、油脂酵母(Lipomycessp.)(例如斯达氏油脂酵母)、聚球蓝细菌属(Synechococcus sp.)、衣藻属(Chlamydomonassp.)(例如莱茵衣藻(C.reinhardtii))、亚罗酵母(例如解脂亚罗酵母)。
在某些实施方案中，宿主细胞与本文所述的天然存在的FAS(亚基)衍生自的细胞是相同的物种。在某些实施方案中，宿主细胞与本文所述的天然存在的FAS(亚基)衍生自的细胞是不同的物种。
一方面，本发明涉及生产如本文所述的重组细胞的方法，其包含以下步骤，将如本文所述的根据本发明的多核酸、载体或编码多肽的多核酸导入到原核或真核细胞，优选如本文他处所述的原核或真核细胞中。本领域已知导入核酸或蛋白质的方法，并包括但不限于转化、转染、脂质体转染、电穿孔、基因枪的使用等。如本文他处所说明，在宿主细胞中导入的多核酸或载体可以保持附加的(即染色体外的)或可以部分或完全地整合到宿主细胞基因组中(定点整合或通过随机整合)，并且其编码的多肽可以组成地、条件性或诱导地表达。
在相关方面，本发明提供了生产重组细胞的方法，其包含以下步骤：
(i)提供本文定义的原核或真核细胞并包含脂肪酸合酶(FAS)基因或多个FAS亚基基因，优选如本文他处定义的FAS(亚基)，任选地具有至少两个酰基载体蛋白(ACP，优选如本文他处定义的ACP)编码核苷酸序列；和
(ii)导入或插入TE序列，或用TE编码序列(优选如本文他处定义的TE)替换所述ACP编码核苷酸序列之一。
该方法可以有利地牵涉如本领域已知的敲入(knock-in)技术的使用。
本发明在另一方面涉及如本文所述的多核酸、载体、多肽或宿主细胞在脂肪酸，例如特别是短链和/或中链脂肪酸的生产中的用途。
另一方面，本发明涉及生产脂肪酸，特别是短链和/或中链脂肪酸的方法，其包含以下步骤：
(i)提供如本文所述的根据本发明的重组原核或真核细胞；和
(ii)在适合的培养条件下培养所述重组原核或真核细胞。
本领域公知培养如本文所述的宿主细胞的方法。技术人员将理解，适合的培养方法包括维持宿主细胞存活和代谢活性，并优选允许宿主细胞的生长和/或繁殖。适合的培养条件包括允许由本文所述的根据本发明的多核酸编码的多肽(即经修饰的FAS或FAS片段，例如FAS亚基(例如FAS1和/或FAS2)，或它们的片段)表达的条件。适合的培养条件还包括提供允许生产脂肪酸，特别是短链和/或中链脂肪酸的必需成分。
在某些实施方案中，如本文所述的生产脂肪酸的方法还包含分离和/或纯化脂肪酸。如本文所用，分离和/或纯化包括从宿主细胞中分离脂肪酸，其可以通过本领域已知的任何手段，例如离心、过滤等完成。进一步纯化步骤可以包括浓缩脂肪酸和/或去除杂质，以及脂肪酸分馏。适合的纯化条件包括例如TLC。
本发明在另一方面也提供了生产碳氢化合物(例如中或短链碳氢化合物)的方法，其包含实施如本文所述的根据本发明的生产脂肪酸的方法，进一步包含还原、氢化、脱羧或脱碳所述脂肪酸以生产烯烃或烷烃。任选地，如本领域中已知的，所述脂肪酸可以在氢化之前酯化以生产碳氢化合物。
本发明在另一方面也提供了生产脂肪醛(例如中或短链脂肪醛)的方法，其包含实施如本文所述的根据本发明的生产脂肪酸的方法，进一步包含如本领域已知的将所述脂肪酸还原为脂肪醛。
本发明在另一方面也提供了生产脂肪醇(例如中或短链脂肪醇)的方法，其包含实施如本文所述的根据本发明的生产脂肪酸的方法，进一步包含如本领域已知的将所述脂肪酸还原为脂肪醛，随后进行醇脱氢酶步骤以将所述脂肪醛转化为脂肪醇。
可以在生产脂肪酸之后，例如在纯化所述脂肪酸之后，实施脱羧、氢化、酯化、还原等步骤，但或者可以与生产所述脂肪酸同时实施，例如通过在宿主细胞中导入适合的脱羧酶、羧酸还原酶、醛还原酶或醇脱氢酶、脱碳酶(decarbonylase)或其他相关酶，或者通过如本文他处所述的手段导入编码适合的脱羧酶、醛还原酶或醇脱氢酶、脱碳酶或其他相关酶的多核酸。技术人员将理解，可以使用本领域已知的其他手段和方法，例如包括非酶促方法，以获得所需的终产物。
现在将通过以下非限制性实施例和附图进一步描述上述公开内容，其中附图显示：
实施例1：FAS编码基因或cDNA的分离
将从YPD培养基(10g/L酵母提取物、20g/L蛋白胨和20g/L葡萄糖)中的1ml圆红冬孢酵母的培养液中收获的细胞沉淀立即在液氮中冷冻并储存在-70℃。遵循制造商的说明使用FastRNA Pro Red试剂盒和FastPrep仪器(Qbiogen,Inc.,Irvine,USA)从约30-50mg细胞样品中分离总RNA，并且FastPrep仪器的设置为6.0m/s 60秒。通过Nanodrop ND1000分光光度计(ThermoFisher Scientific)测定RNA浓度和质量，而通过琼脂糖凝胶电泳评估RNA完整性。通过PrimeScriptTMHigh Fidelity RT-PCR试剂盒(Takara Bio Inc.)合成cDNA。如前所述(Burke,D.,Dawson,D.&Stearns,T.(2000)Methods in yeast genetics:a ColdSpring Harbor Laboratory course manual,Cold Spring Harbor Laboratory Press,Plainview,New York.)提取酿酒酵母和Aplanochytrium kerguelense的基因组DNA。基因特异性引物(表3)用于扩增FAS基因的开放阅读框(ORF)。
实施例2：表达FAS和它的突变体的重组质粒的构建
从酿酒酵母CEN.PK113-11C(MATa SUC2 MAL2-8c ura3-52 his3-Δ1，由P.University of Frankfurt,Germany友情提供)的基因组DNA或先前构建的质粒(Buijs,N.A.,Zhou,Y.J.,Siewers,V.&Nielsen,J.(2015)Long-chain alkane production by theyeast Saccharomyces cerevisiae,Biotechnol Bioeng.112,1275-1279)中扩增含有酿酒酵母的基因启动子和终止子的片段。通过DNA assembler(Shao,Z.,Zhao,H.&Zhao,H.(2009)DNA assembler,an in vivo genetic method for rapid construction ofbiochemical pathways,Nucleic Acids Research.37,e16.和Zhou,Y.J.,Gao,W.,Rong,Q.,Jin,G.,Chu,H.,Liu,W.,Yang,W.,Zhu,Z.,Li,G.,Zhu,G.,Huang,L.&Zhao,Z.K.(2012)Modular Pathway Engineering of Diterpenoid Synthases and the Mevalonic AcidPathway for Miltiradiene Production,J Am Chem Soc.134,3234-3241.)构建用于酵母中FAS复合物的表达的质粒。之前描述的(Heckman KL,Pease LR.2007.Gene splicing andmutagenesis by PCR-driven overlap extension.Nat.Protoc.2:924-932.)重叠延伸PCR方案用于融合多个DNA片段。表4中列出了每个PCR反应的引物和模板。使用含有URA3选择标记的酵母2μ载体pYX212并通过用限制性内切酶SphI和EcoRI消化来线性化。将伴随有线性化载体的这些片段(列于表4中)转化到酿酒酵母CEN.PK113-11C中并在SC-URA平板(6.7g/L无氨基酸酵母氮源基础(ForMedium,Norfolk,UK)、0.77g/L无尿嘧啶的完全补充混合物(ForMedium,Norfolk,UK)、20g/L葡萄糖)上进行选择。通过使用Zymoprep yeast plasmidminiprep II试剂盒(Zymo Research,Orange,CA)提取质粒，并然后转化到大肠杆菌DH5α感受态细胞中。挑取在具有氨苄青霉素的Luria–Bertani(LB)琼脂平板上生长的大肠杆菌菌落并培养用于质粒分离。将来自大肠杆菌的提取的质粒进行限制酶切消化以计算装配保真性。FAS的表达组件如图2所示。
表3：用于构建表达FAS的质粒的引物
表4：用于装配表达FAS基因或ORF的质粒的PCR片段
实施例3：FAS基因的功能验证
通过乙酸锂/单链载体DNA/聚乙二醇方法(Daniel Gietz,R.&Woods,R.A.(2002)Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method,Methods Enzymol.350,87-96.)将表达FAS编码基因或ORF的重组质粒转化到FAS缺陷菌株PWY12(MATα ura3 leu2 his3 trp1 can1 Δfas1::HIS3Δfas2::LEU2,Wenz,P.,Schwank,S.,Hoja,U.&H.-J.(2001)A downstreamregulatory element located within the coding sequence mediates autoregulatedexpression of the yeast fatty acid synthase gene FAS2by the FAS1gene product,Nucleic Acids Research.29,4625-4632.)中。如果需要的话，在培养基中补充0.5mM棕榈酸和0.5mM硬脂酸(5mM TWEEN 80中储备溶液/乙醇＝1:1)。具有脂肪酸的YPD培养基用于培养用于转化的PWY12，在SC-URA+FA培养基平板(6.7g/L无氨基酸酵母氮源基础、0.77g/L无尿嘧啶完全补充混合物、20g/L葡萄糖，含有脂肪酸)上选择转化体。将单菌落接种到“SC-URA+FA”培养基中以扩增细胞24-36小时，然后将细胞快速离心(spin down)并用无菌水清洗两次，并且在“SC-URA”培养基中生长24小时以耗尽脂肪酸残留。之后，将细胞在“SC-URA”或“SC-URA+FA”平板上划线培养以测试脂肪酸自养生物的恢复。互补试验结果如图3中所示。
实施例4：重组FAS的短/中链脂肪酸生产
将表达FAS基因或ORF的酿酒酵母PWY12的重组转化体在具有100mg/L色氨酸的2mlDelft培养基(Jensen,N.B.,Strucko,T.,Kildegaard,K.R.,David,F.,Maury,J.,Mortensen,U.H.,Forster,J.,Nielsen,J.&Borodina,I.(2014)EasyClone:method foriterative chromosomal integration of multiple genes Saccharomyces cerevisiae,FEMS Yeast Res.14,238-248.)中预培养过夜。将细胞接种到100ml Erlenmeyer烧瓶中的20ml“Delft+Trp”培养基中以实现初始光密度(600nm处的OD)为0.1。培养48小时后，取4ml培养液用于细胞外脂肪酸的分析，通过离心收获剩余的细胞沉淀，用Millipore-Q水清洗一次，并冻干用于细胞内总脂肪酸的测量。
开发了一种改进的方法用于细胞外S/MCFA的提取和酯化。简而言之，在提取管(16x100mm培养管和GPI 15-415Threaded Screw Cap,Corning Inc.,US)中的4ml培养液添加0.5ml 10％(w/v)NaCl、0.5ml冰乙酸(含有10ug庚酸和10ug十五烷酸作为内标)和2ml 1:1(v/v)氯仿/甲醇。于1800rpm涡旋30分钟后，将混合物于3000rpm离心至少10分钟，并通过玻璃注射器将下层氯仿相转移到干净的提取管中。通过将1ml三氟化硼/甲醇(14％,w/w,Sigma-Aldrich)与200ul氯仿提取液混合生成脂肪酸甲酯(FAME)，并于室温酯化过夜。这是基于通过三氟化硼/甲醇的游离脂肪酸的即时和快速的酯化(Mitchell,J.,Smith,D.M.&Bryant,W.M.D.(1940)Analytical Procedures Employing Karl FischerReagent.1III.The Determination of Organic Acids,J Am Chem Soc.62,4-6.)。通过添加1ml H2O和600ul己烷，于1500rpm涡旋10分钟并于1000g离心10分钟来提取FAME。通过GC/MS分析取自己烷相的200ul。提取细胞内总脂肪酸并通过稍作修改的先前描述的方法(Khoomrung,S.,Chumnanpuen,P.,Jansa-ard,S.,Nookaew,I.&Nielsen,J.(2012)Fast andaccurate preparation fatty acid methyl esters by microwave-assistedderivatization in the yeast Saccharomyces cerevisiae,Appl MicrobiolBiotechnol.94,1637-1646.)进行酯化。庚酸和十五烷酸用作内标，并且1ml己烷用于提取。
通过使用ZB-WAX柱(30m*0.25mm*0.15um,Phenomenex Inc.,UK)和氦气作为气体载体(3ml/分钟)的Thermo Scientific ISQ单四极杆GC-MS系统(Thermo FisherScientific Inc.,USA)分析提取的FAME。注射1ul样品(不分流，240℃)，并将烘箱温度设置在30℃2分钟；以40℃/分钟的升温速率升高至150℃，保持2分钟；以10℃/分钟的升温速率升高至250℃，保持3分钟。通过与商业标准和NIST质谱数据库的比较来指定化合物身份。FAME的定量基于每个单独标准的校准曲线，并且特定离子(对于饱和FAME为m/z 87，对于单烯FAME为m/z 74)的面积用于定量。将FA的浓度相对于内标的浓度标准化。半定量色谱如图4中所示。细胞外S/MCFA的滴度如图5、7和8中所示，并且细胞内FA组成列于图6中。
实施例5：含有TE的FAS的整合表达
从酿酒酵母基因组扩增URA3基因旁侧的上游和下游区域(约500bp)并用于URA3基因座中FAS基因的整合。从pUG6扩增G418抗性基因盒KanMX(Gueldener,U.,Heinisch,J.,Koehler,G.J.,Voss,D.&Hegemann,J.H.(2002)A second set of loxP marker cassettesfor Cre-mediated multiple gene knockouts in budding yeast,Nucleic AcidsRes.30,e23.)。如实施例2中所述，分别从质粒pScFAS-WT和pScFAS-ACP-TE扩增野生型和杂合体酿酒酵母FAS基因的表达盒。使用表5中列出的PCR片段转化YJZ02(MATa SUC2 MAL2-8cura3-52 his3-Δ1 pox1Δ)，其是通过删除编码过氧化物酶体酰基辅酶A氧化酶(负责脂肪酸β-氧化的酶)的POX1基因衍生自酿酒酵母CEN.PK113-11C的同系菌株)。在“YPD+G418”平板(含有200mg/L G418的YPD培养基)上选择转化体，并使用引物对ID-dURA3-UP/ScFAS2-R1、ScFAS2-F1/ScFAS1-F1和ScFAS1-R1/ID-dURA3-DOWN的菌落PCR验证(Looke M,Kristjuhan K,Kristjuhan A:Extraction of genomic DNA from yeasts for PCR-basedapplications.BioTechniques 2011,50(5):325-328.)。将具有整合FAS的正确转化体在2ml“Delft+His+Ura”培养基(具有100mg/L组氨酸和100mg/L尿嘧啶的Delft培养基)中预培养过夜，并然后将细胞接种到100ml Erlenmeyer烧瓶中的20ml“Delft+His+Ura”中以实现初始光密度(600nm处的OD)为0.1。培养48小时后，如上所述，取4ml培养液用于细胞外脂肪酸的分析。整合地表达FAS的酿酒酵母的滴度如图9中所示。
表5：用于FAS基因或ORF的整合表达的PCR片段
实施例6：重写真菌脂肪酸合酶，用于经调整的化学生产
材料和方法
质粒、菌株和培养条件
通过TA克隆试剂盒(Takara Bio，大连，中国)、限制酶消化/连接方法25、无限制性方法(restriction-free method)26、Gibson装配克隆试剂盒(New England Biolabs，MA，US)或DNA assembler构建所有质粒。大肠杆菌DH5α用于质粒扩增，并且大肠杆菌BL21(DE3)用于重组蛋白表达。如果没有具体说明，则在如有需要补充有适量的抗生素(100mg/L氨苄青霉素和/或50mg/L卡那霉素)的Luria–Bertani(LB)培养基中于37℃和200rpm培养大肠杆菌。衍生自CEN PK113-11C(MATa SUC2 MAL2-8c his3Δ1 ura3-52)的酿酒酵母菌株BY4741(MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0)或YJZ029(MATa SUC2 MAL2-8c his3Δ1ura3-52 Δpox1)用于体内质粒装配。酿酒酵母PWY1229(MATαura3 leu2 his3 trp1 can1Δfas1::HIS3 Δfas2::LEU2)用于互补试验。菌株PWY12和YJZ02用作S/CMFA生产的宿主。“YPD”培养基(10g/L酵母提取物、20g/L蛋白胨和20g/L葡萄糖)用于酵母菌株的常规培养。含有200mg/L G418(Formedium)的“YPD+G418”用于选择具有kanMX盒的转化体。含有20g/L葡萄糖、6.7g/L无氨基酸酵母氮源基础(YNB，Formedium)和0.77g/L无尿嘧啶完全补充混合物(CSM-URA，Formedium)的“SC-URA”培养基用于选择对尿嘧啶原养型的转化体。对于PWY12菌株的培养，将脂肪酸(将含有50mM硬脂酸和50mM棕榈酸的100X储备溶液溶解在Tween 80/乙醇(1:1，v/v)中)添加到培养基中。将20g/L琼脂添加到在固体培养基中。将酵母细胞在液体培养基中于30℃和200rpm培养。通过GENESYS 20分光光度计(Thermo Scientific)测量细胞密度(OD600)。
克隆RtFAS的cDNA
如上所述提取圆红冬孢酵母的总RNA。通过PrimeScript High Fidelity RT-PCR试剂盒(Takara Bio，大连，中国)合成cDNA。通过使用引物FAS1-L1/FAS1-R1从cDNA扩增RtFAS1，并且分别通过使用引物FAS2-69F/FAS2-3021R、FAS2-2660F/FAS2-4529R和FAS2-4304F/FAS2-8881R扩增RtFAS2的三个片段。通过TA克隆将所有这些cDNA片段插入到pMD19-T载体(Takara Bio，大连，中国)中，并通过DNA测序验证。
来自RtFAS的分立的ACP和PPT结构域的表达和纯化
通过基于PCR的方法30生成RtACPI的S1062A突变体(mRtACPI)和RtACPII的S1253A突变体(mRtACPII)。pMD19T-RtFAS2(2660-4529)用作模板，并且引物S1062A-F/S1062A-R和S1253A-F/S1253A-R分别用于RtACPI和RtACPII的定点突变。引物对41-GST-ACPI-F/41-GST-ACPI-R、41-GST-ACPII-F/41-GST-ACPII-R和41-GST-PPT-F/41-GST-PPT-R用于扩增RtACPI、RtACPII和RtPPT片段，将所述片段如前所述通过无限制性克隆插入到pET-41(a)载体(Novagen)中。
将4ml具有质粒(表达GST-wRtACPI、GST-wRtACPII、GST-mRtACPI、GST-mRtACPII或GST-RtPPT)的大肠杆菌BL21(DE3)过夜培养物接种到400ml补充有50μg/ml卡那霉素的Terrific Broth(TB)培养基(每升12g胰蛋白胨、24g酵母提取物、4ml甘油、2.31g KH2PO4和12.54g K2HPO4)中并于37℃生长。当OD600为约0.8时将1mM IPTG添加到培养物中，然后细胞于20℃生长24小时。通过离心收获细胞，并将4g湿物质悬浮于24ml补充有20mM咪唑和1mg/ml溶酶菌的NBP缓冲液(50mM Na2HPO4/NaH2PO4，pH 8.0，0.5M NaCl，1mM 2-巯基乙醇，1mM PMSF)中。将悬浮液在冰上保持30分钟，然后通过超声波破碎。离心(12000rpm于4℃20分钟)用于去除细胞碎片。将上清液与Ni-NTA琼脂糖(Life Techologies)混合并在冰上平衡15分钟。然后由具有20、40、60和80mM咪唑的NBP缓冲液依次清洗树脂并由含有250mM咪唑的NBP缓冲液洗脱。用Amicon Ultra-15离心过滤单元(MWCO 10kDa，Millipore)浓缩洗脱的蛋白质，并将缓冲液变为磷酸泛酰巯基乙胺化反应缓冲液(20mM Tri-Cl，pH 7.5，10mMNaCl，100mM KCl，5mM MgCl2，10mM CaCl2，1mM 2-巯基乙醇，0.5mM DTT，15％甘油)，然后将蛋白质储存在-20℃。
ACP的体外磷酸泛酰巯基乙胺化
将含有20μL反应混合物(20μg GST-RtACP、2μg GST-RtPPT和0.3mMCoA于磷酸泛酰巯基乙胺化反应缓冲液中)的200μL管在热循环仪(Eppendorf,Germany)中于30℃温育3小时。将反应产物在由MALDI-TOF/TOF5800质谱仪(Applied Biosystems,Framingham,MA,USA)在正线性模式下分析之前储存在-20℃。该仪器配备有355nm处的Nd:YAG激光，重复频率为400Hz。优化激光能量范围以获得良好的信噪比(S/N)。通过使用标准蛋白质实施外部质量校准。为了分析蛋白质，将样品沉积在MALDI靶上，并然后添加1μL基质溶液(在含有0.1％TFA的50％ACN中20mg/ml的芥子酸)用于MS分析。或者，将2.5μL肠激酶(1U，SangonBiotech，上海，中国)添加到10μL反应混合物中并于25℃温育16小时。由16％Tricine-SDS-PAGE(丙烯酰胺:双丙烯酰胺＝29:1)31分析经切割的蛋白质。
RtFAS复合物和它的突变体的表达和纯化
由所述方法在酿酒酵母BY4741中装配RtFAS基因，并且2μ附加型载体pYX212用作骨架。通过引物对FAS1-5-NdeI/FAS1-3-EcoRI扩增RtFAS1的完整ORF。用NdeI/EcoRI消化片段并将其插入到pET22b(+)(Novagen)中以生成pET22b-RtFAS1。通过引物对FAS2-5-HindIII/FAS2-3-NotI扩增RtFAS2的完整ORF和它的两个突变体(S1062A或S1253A)，将其用HindIII/NotI消化并插入到pET24b(+)(Novagen)中以分别生成pET24b-RtFAS2、pET24b-RtFAS2(S1062A)和pET24b-RtFAS2(S1253A)。将用于RtFAS1和RtFAS2两者表达的质粒同时转化到大肠杆菌BL21(DE3)中并在具有100mg/L氨苄青霉素和50mg/L卡那霉素的LB平板上进行选择。之前描述了RtFAS复合物的纯化17、22，并且可以通过三步纯化(硫酸铵沉淀、蔗糖密度梯度离心和阴离子交换色谱)获得相当均一的蛋白质复合物。通过监测丙二酰-CoA和乙酰-CoA依赖性NADPH氧化来测定脂肪酸合酶活性。于室温(约25℃)和200μL石英比色皿中实施测定，所述比色皿中含有0.1M磷酸钾(pH 7.0)、5mM DTT、12.5μM乙酰辅酶A、50μM丙二酰辅酶A、75μM NADPH和大约10μg经纯化的酶。当监测没有丙二酰辅酶A的空白反应时，在340nm处的吸收没有明显减少。并且活性(1U)定义为每分钟1μmol NADPH的转换。
质粒构建和基因操作
分别从pET22b-RtFAS1和pET24b-RtFAS2扩增RtFAS1和RtFAS2。从A.kerguelense的基因组DNA扩增AkFAS，从酿酒酵母基因组DNA扩增ScFAS1和ScFAS2基因、启动子和终止子片段。来自pYX212的2μ起点用作复制元件，并且来自pYX212的ScURA3或来自pWJ104232的KlURA3用作选择标记。通过Genscript合成密码子优化的基因(AcTesA、ShMKS1和ShMKS2，补充表S7)。将所有这些质粒转化到FAS缺陷菌株PWY12中，并测试了它们对PWY12的脂肪酸营养缺陷型的互补作用。转化之后，“SC-URA+FA”平板用于选择转化体。将单菌落接种到“SC-URA+FA”液体培养基中以扩增细胞24-36小时，然后将细胞快速离心并清洗两次，并在“SC-URA”液体培养基中生长24小时以耗尽脂肪酸残留。之后，将细胞在“SC-URA”和“SC-URA+FA”平板上划线培养或接种到“SC-URA”液体培养基中以测试脂肪酸自养生物的恢复。含有空载体(pYX212)的PWY12不在“SC-URA”培养基中生长。FAS表达盒的整合也是根据先前所述的方法。使用了ura3-52基因座上游和下游约500bp同源臂和来自pUG633的KanMX选择标记。将ScFAS01、ScFAS27、ScFAS15和ScFAS28整合到YJZ02的ura3-52基因座中以分别生成菌株ZW201、ZW202、ZW206和ZW207。通过Gibson装配构建pZWM1-AcTesA质粒用于'AcTesA的附加型表达。
代谢物提取和定量
在代谢物提取之前，如果没有具体说明，细胞在具有20ml根据需要补充有成分(100mg/L尿嘧啶、100mg/L组氨酸和/或100mg/L色氨酸)的Delft培养基的100ml摇瓶中生长48小时。初始光密度(600nm处的OD)为0.1。通过具有修改的前述方法10提取和酯化细胞外S/MCFA。简而言之，在提取管(16x100mm培养管和GPI 15-415Threaded ScrewCap,Corning Inc.,US)中的4ml培养液中添加0.5ml 10％(w/v)NaCl、0.5ml冰乙酸(含有10μg庚酸和10μg十五烷酸作为内标)和2ml 1:1(v/v)氯仿/甲醇。于1800rpm涡旋30分钟后，将混合物于3000rpm离心10分钟，并通过玻璃注射器将下层氯仿相转移到干净的提取管中。通过将1ml三氟化硼/甲醇(14％，Sigma-Aldrich)与200μL氯仿提取液混合生成脂肪酸甲酯(FAME)，并于室温酯化过夜。这是基于通过三氟化硼/甲醇35的游离脂肪酸的即时和快速的酯化。然后通过添加1ml H2O和600μL己烷，于1500rpm涡旋10分钟并于1000g离心10分钟来提取FAME。通过GC/MS分析取自己烷相的200μL。对于高产S/MCFA的菌株(ZW201、ZW202、ZW206和ZW207)，使用将1ml培养液稀释于3ml Milli-Q H2O中。
根据稍作修改的先前描述的方法微波辅助酯化酵母生物质后，提取细胞内总脂肪酸(以FAME的形式)。将1ml己烷、2ml三氟化硼/甲醇(14％，Sigma-Aldrich)、10μg庚酸和10μg十五烷酸添加到10mg冻干生物质中用于总脂肪酸的衍生化，在微波辅助酯化后，取上层己烷相中的FAME进行GC/MS分析。
通过使用ZB-50柱(30m*0.25mm*0.25um,Phenomenex Inc.,UK)的FOCUS GC/ISQ单四级杆质谱仪系统(Thermo Fisher Scientific Inc.,USA)分析提取的短/中链FAME。氦气用作气体载体(3ml/分钟)。注射1μL样品(不分流，240℃)，并将烘箱温度设置在30℃2分钟；以40℃/分钟的升温速率升高至150℃，保持2分钟；以10℃/分钟的升温速率升高至250℃，保持3分钟。然而，对于长链FAME，使用ZB-WAX柱(30m×0.25mm×0.25um,Phenomenex Inc.,UK)，并且初始烘箱温度为50℃。MS传输线和离子源的温度分别设置为250℃和200℃。以全扫描模式(50-450m/z)和选择离子监测模式(74m/z)来检测衍生自电子电离(70eV)的碎片离子。特定离子(m/z 74)的面积用于FAME的定量。
结果
真菌I型FAS复合物是一个桶形中空颗粒，由在赤道中心轮的分隔以形成两个反应室(图11b)。真菌FAS由一个或两个多肽编码，并通常含有七个催化酶结构域和一个充当蛋白质辅因子的ACP结构域(图11b和11c)。通过磷酸泛酰巯基乙胺基转移酶(PPT)催化的假体磷酸泛酰巯基乙胺基团的附着激活ACP，其提供巯醇基团以连接生长的脂肪酰链。ACP结构域位于隔室内，并由两个固定在室壁和中心轮上的柔性接头拴系(图11b)。携带酰基货物的ACP动态地与其他催化结构域相互作用以实现完整的反应循环。由于ACP和它的相邻的接头在中空反应室中是可移动的，我们推测这些柔性区域可以很容易地得到修饰，并因此可以将利用酰基-ACP作为底物的异源酶整合到FAS中以产生新的可以生产多种脂肪酸和脂肪酸衍生的化学物质的合成FAS机器。(图11b和11c)。含油酵母圆红冬孢酵母的先前基因组注释揭示了特殊的FAS，其根据它们的蛋白质结构域构造与来自其他真菌物种的FAS不同(图11a)。然而，来自圆红冬孢酵母的FAS(RtFAS)装配成由cryo-EM分析揭示的典型真菌FAS结构。更有趣的是，RtFAS包含两个彼此高度相似的推定ACP结构域。(对来自真菌FAS的ACP结构域实施序列比对。序列包括来自酿酒酵母的Fas2的ScACP(Genbank登录号，P19097.2)、来自新型隐球酵母的Fas1的CnACP(Genbank登录号，XP_571100.1)、来自玉米黑粉菌的Fas的UmACP(Genbank登录号，XP_759118.1)、来自A.kerguelense的Fas的AkACPI和AkACPII(JGI蛋白质ID,103951)、来自P.graminis的Fas2的PgACP(Genbank登录号，XP_003889657.1)、来自圆红冬孢酵母的Fas2的RtACPI和RtACPII(Genbank登录号，EMS21268.1)、来自C.reversa的Fas的CrACP(JGI蛋白质ID,10860)；数据未显示)。
当使用分立的ACP和PPT结构域(作为经纯化的GST融合蛋白)时，我们发现两个ACP都可以通过来自RtFAS的同源PPT进行体外磷酸泛酰巯基乙胺化。还显示ACP的磷酸泛酰巯基乙胺化依赖于辅酶A、PPT酶和磷酸泛酰巯基乙胺附着位点中的保守的丝氨酸残基(图12和表6)。
表6：通过MOLDI-TOF质谱法测定ACP的分子量。在磷酸泛酰巯基乙胺化期间，ACP的分子量增加340Da。
我们已经建立了对于大肠杆菌中重组RtFAS复合物的纯化方案，从其中可以通过连续(NH4)2SO4沉淀、蔗糖密度梯度超速离心和DEAE-琼脂糖阴离子交换色谱获得稳定并有活性的FAS复合物。为了研究重复的ACP在脂肪酸合成中的作用，将两者中任一ACP的磷酸泛酰巯基乙胺附着位点中保守的丝氨酸残基突变为丙氨酸残基。通过监测NADPH氧化对经纯化的FAS复合物的酶测定显示两个突变体(S1062A和S1253A)具有与野生型相当的脂肪酸合成活性(图13)。一致地，含有ScFAS1和ScFAS2缺失的酿酒酵母PWY12与野生型RtFAS(RtFAS01)和突变形式的RtFAS(S1062A,RtFAS02和S1253A,RtFAS03)的互补(图14a和14b)，以及与野生型RtFAS01相比，由突变体(RtFAS02和RtFAS03)生产的几乎相同量的脂肪酸(图14b和表7)，说明两个ACP在脂肪酸合成中起类似作用，并且一个单一ACP对于RtFAS的活性是足够的。
表7：由表达RtFAS的PWY12和它的突变体生产的细胞内总脂肪酸。
单位，mg/g DCW。数据来自三个独立培养物，并且显示了平均值±SD。
我们还发现了一些来自不同系统发育进化枝的含有重复的ACP的真菌FAS。类似地，来自Aplanochytrium kerguelense的由一个单一多肽编码的FAS(AkFAS01)和它的含有一个有活性的ACP的两个突变体AkFAS02(S2150A)和AkFAS03(S2340A)分别在酿酒酵母PWY12中得到表达(图14a)。其也表明，AkFAS中只有一个ACP对于脂肪酸合成是足够的(图14c)。
在证明重复的ACP在真菌I型FAS中的冗余作用后，我们考虑用短链酰基-ACP硫酯酶(sTE)替换RtFAS和AkFAS中的任一ACP以水解酰基-ACP中间体并生产S/MCFA，其是许多工业化学品的前体，并且还与改进生物燃料的性质有关。将来自Acinetobacter baylyi的优选生产S/MCFA23的独立式(free-standing)硫酯酶(’AcTesA)插入到FAS中(图15a)。根据真菌FAS的结构，嵌入的sTE将定位在反应室中并容易接近酰基-ACP底物。从互补试验，我们发现含有sTE的FAS(RtFAS04、RtFAS05、AkFAS04和AkFAS05)可以合成必需脂肪酸以支持对脂肪酸是营养缺陷型的酿酒酵母PWY12的生长(补充图15a-c)。然后我们提取并定量了由表达不同FAS变体的酿酒酵母PWY12生产的细胞外脂肪酸，并发现与相应的野生型RtFAS和AkFAS相比，杂合体FAS生产5-10倍更多的细胞外S/MCFA(图15b和15c)。特别是表达RtFAS05的菌株生产了0.67mg/L己酸和0.72mg/L辛酸，其比野生型RtFAS01生产的多约20倍(图15b)。有趣的是，RtFAS04和RtFAS05与RtFAS01相比分别生产了约20％和40％更少的C18脂肪酸，显示嵌入的sTE确实干预了酰基链延伸(表7)。
RtFAS的EM结构与棉毛状嗜热霉(Thermomyces lanuginosus)和酿酒酵母FAS的X射线结构很好地重叠，表明这些FAS具有相似大小的反应室。当用来自RtFAS的串联双ACP替换来自酿酒酵母的FAS(ScFAS)的单一ACP时，含有两个ACP的杂合体ScFAS09保留了脂肪酸合成活性(图15)，表明真菌FAS的反应室可以容纳更多内容物。类似地，含有一个有活性ACP和一个失活ACP的ScFAS10和ScFAS11补充了酿酒酵母PWY12中的FAS缺失。这使我们通过邻近ACP结构域插入sTE来构建类似的杂合体ScFAS(ScFAS14和ScFAS15)(图1a)。我们发现与野生型ScFAS01相比，ScFAS14和ScFAS15生产更高水平的S/MCFA。为了进一步说明嵌入的sTE是否在反应室中起作用并水解酰基-ACP而不是酰基辅酶A，其由丙二酰/棕榈酰转移酶(MPT)释放并然后通过FAS颗粒壁中的开窗扩散到中空腔中，将具有嵌入的sTE的FAS(ScFAS14和ScFAS15)的S/MCFA生产与具有外围的或游离的sTE的FAS进行比较。如果这些FAS生产相似量的S/MCFA，则酰基辅酶A用作ScFAS14和ScFAS15中sTE的底物，而如果它们两者都比具有外围的或游离的sTE的FAS生产更多的S/MCFA，则酰基-ACP用作ScFAS14和ScFAS15中sTE的底物。因此，将sTE与PPT的C末端融合，以将外围的sTE附着于FAS颗粒。ScFAS16补充了酿酒酵母PWY12中的FAS缺失，表明该温和的修饰没有太多干扰FAS结构。游离的sTE也与ScFAS(ScFAS17)共同表达。我们发现由ScFAS14和ScFAS15生产的S/MCFA比ScFAS16和ScFAS17生产的多60-210％，其中sTE对FAS室中拴系的酰基-ACP底物的接近有限(图15d和图16)。此外，与高拷贝质粒的sTE高水平表达相比，ScFAS15(单拷贝)的整合表达生产2倍更多的S/MCFA(图17a)。这些结果清楚地显示，ScFAS中的嵌入的sTE比外围或游离的sTE更好地作用于ScFAS对于S/MCFA生产。
序列表
<110> 道达尔炼油化学公司
<120> 生产脂肪酸的方法
<130> TOTAL-213-PCT
<150> EP16158805.8
<151> 2016-03-04
<160> 108
<170> PatentIn version 3.5
<210> 1
<211> 3801
<212> DNA
<213> 圆红冬孢酵母
<400> 1
atgaacggcc gagcgacgcg gagcgtgact gggacgtcga cgccggtcca cacggcgacg 60
acccgacccc tcgtcctctt gcacccctcg acccaaaccc gcatctcgct gcacgtcccc 120
tccacgtcgc aggaatggat cgccgccgaa gtcgcgcgcg acaccttcca ggactggctt 180
cacgctgccg agaagagcgg aaacctcgtc ggattcgagg cggccgagct tgacgacgag 240
caggctggcg agggcgacga cgagaaggag ctcgtcctca ccgcctactt cttgaagcac 300
gttgccggcc ttctcccctt cccgtcgaca gctacctccc ccgccaccgc cgccgtcctc 360
ctcgccgcct tcaaccactt tgcgtccgtc tacctcagcg gaaccgatgt tcacaccctc 420
actgcctcgc tcgctgctcc cgtccgcgct ctcgtcatct cgtccttctt cctcgccaag 480
accaagctcg aggtcgaggg actcggcaag gtcttgccca agcagtccga gtcggcgctc 540
ctgcagaagg ctgcgaccgg ccaggcagag gtcttcgctc tcttcggtgg tcagggaatg 600
aacgaggtct actttgacga gctccagacc ctccacgacc tttacacccc gctgcttacg 660
cccttcctcg cccgcgcctc cgaacacctc gtctctctcg ctgccgccga gcagcacacc 720
ctcctttacg accactcgct cgacgccctt gcctggctgc aagatccctc tacccgcccc 780
gaagtcccct acctcgcgac ttgcgccgtc tcgctccctc tcatcggtct cactcagctc 840
tgccagtacg tcgtgtacgg caagggctcg tcgctcggtc ccgccgagct cggcgccaag 900
ttcaagggcg cgaccggcca ctcgcagggt gtcgtctcgg ctcttgtcat cgcgcacgag 960
taccctcccg cgtccaagga cggcagcgac gcgtgggagc ctttctacga gcaggccctt 1020
cgcggtttga ccgtcctctt ccagatcggt ctccagggca cgctcgcctt cccctccatc 1080
gccatttcgc ccgctctcga gtcgagctcg gtcgagaatg gcgagggtgt cccgactgcc 1140
atgcttgccg tcaccggcct cgacctcaag tcgctcgaga agaagatcgc cgaggtcaat 1200
gggcacgtca agtctgaggg ccgcgacgag accgtctcga tcagtctcta caacggtgcg 1260
agggcgttcg tcgtcactgg tgcgccgaag gacctcgtcg gtctcgccga cggccttcgc 1320
aagaaccgcg cgccggccgg caaggaccag tcgaagatcc cgcactcgaa gcgtctcccc 1380
gtcttctcga tgcgcttcct ccccatcaac gttccctacc actcgcatct cctccaaggc 1440
gcgaccgaga aggcgctcgc gacgttctcg gctgaggagg ccgcccactg ggcgccttca 1500
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tcgtcggttc tcgagtcggt cttccagcag atcttcacct cgcccattca ctgggtctcg 1620
cacgccacca acttcccctc gtccgcgacg cacgccatcg atttcggcac gggcggcgcg 1680
agcggcatcg gttcgctctg cgcgcgcaac tgggagggcc gcggtatccg cacgattatg 1740
ctcggcaacc gcggcgaggg cgttggtgcc ggcaaggagg cttggggcaa gaaggtcccg 1800
accgaggaga agtggaacga gcgcttccac cctcgcctcg tccgcaccag cgacggcaag 1860
atccacctcg acacgccctt ctcgcgcctc ctctcgaagc cgcccctcat ggtcggtggt 1920
atgaccccga cgaccgtcaa ggccggcttc gtctcggccg ttctccgcgc gggctaccac 1980
atcgagctcg ctggcggcgg tcactacaac gagaaggctg tccgtgccaa ggtcgccgag 2040
atccagaagc tcgtgaacaa gcccggcatg ggcatcaccc tcaactcgct ctacatcaac 2100
cagcgccagt ggacgttcca gttcccgctc tgggccaaga tgaagcagga gggcgagccc 2160
gtcgagggtc tctgtgttgc tgccggtatt ccctcaaccg agaaggccaa ggagatcatc 2220
gacacgctcc gcgaggccgg catcaagcac gtctcgttca agcccggttc ggtcgacggc 2280
atccgccagg tcgtcaacat cgcctccgcc aaccccgact tccccatcat cctccagtgg 2340
actggtggtc gcgccggcgg tcaccactcg tgcgaggact tccacgcccc gatcctcgcg 2400
acgtacgctt cgatccgtca gcaccccaac atcaagctcg tcgccggctc tggcttcggc 2460
tcggctgagg gatgctaccc ttacctttcg ggcgagtggt cggagaagca gtacggcgtc 2520
gcgcgcatgc cgttcgacgg cttcatgttt gcttcgtggg tcatggtcgc caaggaggcg 2580
cacacgagcg agtcggtcaa gcagctcatc gtcgacgcgc ctggtgtcga ggatggccag 2640
tgggagcaga cgtacgacaa gccgaccggc ggcatcctca ccgtcaactc ggagcttggc 2700
gagccgatcc acaaggtcgc gactcgtggt gtcaagctgt gggccgagtt cgacaagaag 2760
gtcttctcgc tgtcgaagga gaagcagctc gcatggctcg ccgacaacaa gaagtacgtt 2820
atcgaccgcc tcaacgccga tttccagaag ccctggttcc ccgccaaggc cgacggctct 2880
ccttgcgacc ttgccgacat gacctacgcc gaggtcaacg cccgcctcgt ccgcctcatg 2940
tacgtcgcgc acgagaagcg ctggatcgac ccgtcgctcc gcaacctcgt cggcgactgg 3000
atccgccgtg ttgaggagcg tctctcgaac gtcaacgact cgggcatcaa gatctcggca 3060
ctccagtcgt actcggagct gaacgagcct gaggcgttcc tcaagcagtt cctcgcccag 3120
tacccgcagg ccgaggacca gatcctcgcc tccgccgacg tttcctactt cctcgccatc 3180
tctcaacgcc ccggacagaa gcccgtcccc ttcatccccg tcctcgacgc caacttcagc 3240
atctggttca agaaggactc gctgtggcag gccgaggaca tcgaggccgt ctttgaccag 3300
gacccgcagc gtgtctgcat cctccaggga ccggtcgccg ccaagcactg cacctcgacg 3360
cagacgccca tcgccgagat gctcggcaac atcgagcacc agctcgtcaa gaacgtcctg 3420
gacgactact acggcggcga cgagtcccag atcccgacta tcgactacct cgcgccccct 3480
cccaagccgg tcgacgccgg cgctatcctc gccgagaaca acatcgcgca ctcggtcgag 3540
gagctcgccg acggcggcaa gaagcatgtc tactcgatca acggtgtcct cccgccgacg 3600
ggcgactggc atgccgcact cgccggcccc aagctcgact ggctccaggc gttcctctcc 3660
aacgtctcga ttcaggcggg cgagcagtcg attcctaacc ccgtcaagaa ggtgctggcg 3720
ccgaggcacg ggcagcgggt cgagctcacc ctgaacaagg acggccagcc cctcaagctc 3780
gacgtcttcg gcgggctctg a 3801
<210> 2
<211> 8787
<212> DNA
<213> 圆红冬孢酵母
<400> 2
atggtcgcgg cgcaggactt gccgctcgcg ctgagcatca gcttcgcgcc cgagtcgtcg 60
accatctcga tgacgctgtt caaccagccc gaggcgtcga aacccgccct ccccctcgag 120
ctcaagtaca agtacgaccc ctcgacgccg tacgccccga tccacgagat caccgaggac 180
cgtaatcaga ggatcaagca gcactactgg gacctctggg gcctcggcaa caaggcagac 240
cagggcatct cgcagctcaa gatcaccgac gagttccagg gcgacctcgt caccatctcg 300
gccgacgaga tcgaggcgtt ctgccgtgtt gtcggcatcg agggcgaggc gtacaagcgc 360
aaccacaagg ccggcatgca ggtcccgctc gacttcgcca tcaagctcgg ctggaaggcc 420
atcatgaagc cgatcttccc ctcgacgatt gacggcgacc tgctcaagct cgtccacctc 480
tcgaacggct tccgcgtcct ccccgacacg cccacactcc aggttggcga cgtcgtgacg 540
accacgtcgc gcatcgaatc aatcacgaac tcggacacgg gcaaaaccgt ctcggttcgc 600
ggcgtcatct cgctcgtctc gtccgccgac tcgaagggca aggacgcctc gaccgaggac 660
cgcatcccgc tcatcgaggt cacctcgtcc ttcttctacc gcggcaagtt cagcgactac 720
gcccagacat tctcccgcgt cgcccacccg acctactctg tcccgatcac cacgcccgag 780
gccgtcgccg tcctccagtc caaggagtgg ttccagtggg acgacgactc gaagcccctc 840
gaggtcggca ccaagctcca gttcaaggtc gagtcgaact atgtctacgc cgacaagtcg 900
tcctacgcga tggctaccgt caccggcggc gcgtacgtca tcacccccga gctcaagctc 960
gctgtcaagg ttgccacggt cgactacacg tccgagggcg agggcgtcat ccagggcgac 1020
ccggtcatcg agtacctcaa gcgccacggc tcggccctcg accagcccat catgctcgag 1080
aacggcggct attcgctcac caaggccggc cagtgcacct tcacgacgcc cgcgtccaac 1140
ctcgactact cgctcacctc gggcgacacg aacccgattc acacgaaccc gtactttgcc 1200
tcgctcgcct acctccccgg caccatcacg cacggcatgc actcgtcggc ccgcacgcgc 1260
aagtttgtcg agcaggtcgc cgcagacaac gtcggcgcgc gcgtccgcaa gtacgaggtc 1320
ggcttcacgg ccatgtgcct cccctcgcgc aagatggagg tccgccttaa gcacgtcggc 1380
atgaccgcgg acggaaaccg cctcatcaag gtcgagaccg tcgacgtcga gggcggcaac 1440
gtcgttctca gcggaaccgc cgaggtcgcc caggctccca ccgcgtacgt cttcaccggt 1500
caaggttcgc aagagcccgg catgggcatg gagctctacg ccaactcgcc cgtcgcccgc 1560
gccgtctggg acgaggctga ccgccacctc ggcgaggtct acggcttctc catcctcgag 1620
attgtccgta cgaaccccaa ggaaaagact gtgcacttcg gcgggttgaa aggccaagca 1680
acccgtcaga agtacatgga catgtcgtac acaacgactg accatgaggg caacgttaag 1740
actctcccgc tcttcggcga catcgacctc cgtacctcac gctacacgtt ctcgtcgccg 1800
accggtctcc tctacgccac ccagttcgcc cagatcgccc tcgtcgtaac ggagaaggcc 1860
gccttcgagg acatgcgcgc caagggtctc gttcagaagg actgcgtctt tgccggtcac 1920
tcgctcggag agtactcggc tctcgcctcg atcgccgaca tcctccccat ctcggccctc 1980
gtcgacgtcg tcttctaccg cggtatcacc atgcagcgcg ccgtcgaacg cgaccacctc 2040
aaccgctcgt cgtacggaat ggtcgccgtc aacccgagcc gcatcggcaa gagctttggc 2100
gacgccgccc tccgcgaggt cgtcgacacc atcgcccgcc gcggaaacat cctcatcgag 2160
gtcgtcaact acaacgtcga gggacagcaa tacgtcgtcg ccggtcacct cgtcgccctc 2220
caatccctca caaacgtcct caacttcctc aagatccaga agatcgacct cgccaagctc 2280
accgagacga tgtcgatcga gcaggtcaag gagcacctgt gcgagatcgt cgacgagtgc 2340
gtccagaagg cgcgcgacct ccaggccaag acgggcttca tcaccctcga gcgcggcttt 2400
gcgacgatcc cgctccccgg tatcgacgtg ccgttccact cgcgctacct ctgggcggga 2460
gtcatgccgt tccgcactta cctctcgaag aaggtcaacc cggcgcactt caacgccgac 2520
ctcctcgtcg gccgctacat ccccaacttg accgccgtcc actacgaggt ctcgaaggag 2580
tacgccgaac gcatccacac ccagacgtcg tcgccgcgcc tcaacaagat tctcaaggcc 2640
tgggacgagg agcgctgggg cgcacccgag aaccgcaaca agctcggcta cgccatcctc 2700
atcgagctcc tcgcgtacca gttcgcctcg cccgtccgct ggatcgagac gcaggacatc 2760
ctcttccgcg acttcaagtt tgagcgcctc gtcgagcttg gcccgtcgcc cactctcacc 2820
ggcatggcta cgcgcacgca gaagctcaag tacgacgcgc acgactcgtc ggtcggcatc 2880
aagcgctcga tctactgcat cgccaagcac cagaaggaga tctactacca gttcgatgac 2940
gttgccggcg aagaggcgcc cgctcctgcc gcagttgcgc cttccgctcc cgctcccaag 3000
gccgccccag tcgccgccgc ccctccccct cccgctcctg tcgctgccgc gcctgccgcc 3060
gccgtcgccg acgagccgct caaggctgtc gacacgctcc gcatcatcat cgcgcagaag 3120
ctcaagaagc ccgttggcga agtccccctc accaagtcga tcaaggagct cgtcggcggc 3180
aagtcgaccc tccagaacga gattctcggc gaccttcaag gcgagttcag cagcgcgcct 3240
gaaaagggcg aggagatgcc tctccaggag ctcggcgcgg ccctccagca gggctactct 3300
ggcaagctcg gcaagtacac caccggcgtc atctcgcgca tgattggcgc caagatgccc 3360
ggcggttttg gtctctccgc cgtccagggt cacctcggca agacctacgg cctcggcgcc 3420
ggtcgcatcg atggcgtcct cctcttcgcc gtcacgcagg agccggctaa gcgtctcgcc 3480
aacgagggtg aggcgaaggc ttgggtcgac tcggtcgcgc aaggctacgc ctcgatggct 3540
ggcatctcgc tcgccgccgg cggtggagct gctgctgctg cccccgcgat ggcgttcgcc 3600
gctccggccg cagctggcgg tggagcgccc gctgccgtcc ccgacgagcc gctcaaggcg 3660
accgacacgc ttcgcgccat catcgctcag aagctcaaga agcagatccc cgacgtcccc 3720
ctcaccaagt ccatcaagga ccttgtcggc ggcaagtcga ccctgcagaa cgagatcctc 3780
ggcgacctcc agggcgagtt cagcagtgcg cccgagaagg gcgaggagat gccgctccag 3840
gagcttggcg ccgcactcaa ccaaggctac tcgggcacgc tcggcaagca cacgagcggt 3900
ctcgtcgccc gcatgatggg cgccaagatg cccggtggct tcggtctctc ggcggcgaag 3960
gcgcacctct cgaaggctca cggtctcggg cccggccgca ccgacggcgc tctcctcgtc 4020
gcgctcacca aggagcccga gaaacgtctc ggtagcgagg ccgacgccaa ggcctggctc 4080
gacggcgtcg ctcaggcgta cgcctcgcag gctggcatca ccctcggcgc tggtggaggc 4140
ggaggcggcg cggctgtcgg cggcgccggc tttatgatca acaccgagca gctcgacaag 4200
atgcaggaga agcaggacaa cttcgtctcg cagcaggtcg agctcttcct ccgctacctc 4260
ggcaaggact cgcgcgaggg ccaccgcctc gccgacatgc agaaggcaga ggtcgccaac 4320
ctccaggaga agctcgactc gatcgctcgc gagcacggcg acgcctatgt ccagggcatc 4380
cagcccgtct tcgacccgct caaggcccgc cacttcaact cgtcgtggaa ctgggtccgt 4440
caggacgcgc tcatgatgtg gatggacatc ctcttcggcc gcctcaccac cgtcgaccgc 4500
gacatcaccg ctcgctgcct tgtcatcatg aaccgcgccg acccttctct catcgactac 4560
atgcagtaca ccatcgacaa cacccccgtc gagcgcggcg agcattacgt cctcgccaag 4620
caattcggcc agcagctcct cgacaactgc cgcgagatga tcggccaggc tccgctctac 4680
aaggacgtca ccttcccgac cgcgcccaag acgaccgtca acgccaaggg cgacatcatc 4740
accgaggagg tcaaccgccc cggcgtctct cgcctcgaga agtatgtcgc cgagatggct 4800
gccggctcaa aggtcaccgt cgccagcgtc aacctcgaca aggtccagga gcaggtcgag 4860
aagctgtaca agctcgtcaa gtcgcagccg cagatttcga agcagcacat gacgtcgatc 4920
aagtcgctgt acgctgaggt cgttcgcggt ctcggcaagg acgccggccc tcctccggtc 4980
cacaaggccg gcactcgcgc ccgccgcccc tcgagccagt tcctccgtcc cgcagccgtc 5040
tccgaggcga ctttcctccc cgaggacaag gtgcctctcc tgcacctcaa gcgcaagatc 5100
ggcaacgact ggcaatactc gagcaagctc acgtcgctct acctcgacat cctcaaggag 5160
attgccacgt cgggtgtcac cttcgagcac aagaacgcgc tcatgaccgg tgtcggcaag 5220
ggctccatcg gtatcgagat cgtcaagggt ctcctcgctg gtggcgctcg cgtcgtcatc 5280
acgacctcgc gctactcgcg ctcgactgtc gagtactacc aggcgatcta ccaggaggtc 5340
ggctcgaagg gctcgtcgct caccgtcgtc cccttcaacc agggctcgaa gcaggatgtc 5400
gaggcgctcg tcgacttcat ttattcgaag gataagggtc tcggcatgga cctcgactac 5460
atcctcccct tcgccgccct tcccgagaac ggccgcgaga tcgacggcat cgacgaccgc 5520
tccgagctcg cccaccgcat catgctcacc aacctcctcc gcctcctcgg tgccgtcaag 5580
tcgaagaagg ccgccctcaa gctcacgacc cgcccaaccg aggtcgtcct cccgctttcg 5640
ccgaaccacg gcctcttcgg caacgacggt ctctactcgg agtcgaagat ctcgctcgag 5700
acgctcttca accgctggag ctcggagagc tggggcgagt acctctgcct cgctggcgct 5760
gtcatcggat ggacgcgcgg taccggtctc atgtcggcga cgaactcggt cgccgaaggt 5820
atcgaggcgc agggttgcag gacgttctcc gccaaggaga tggccttcaa cattctcggc 5880
ctcatgcacc cgctcgtctt cgacgtcgcg cagatcgagc ctgtctgggc cgacctcaac 5940
ggtggcatgg acaagctccc cgaccttgcc aacctcacga ccgagatccg caagaagctc 6000
aacctcaccg cgtcgacccg ccgcgccatc gccaaggaca actcgttcga ctacaaggtc 6060
gcgcacggcc cggcgatgga gcagatacac cagcggatca acgtcgcccc gcgcgccaac 6120
ttctcccttc ccttccccga gctcaagccg atcgatgcca agtcggagct cgcgaagctc 6180
cgtggcctca tcgacctcga gaaggtcgta gtcatgaccg gttacgccga ggtcggaccg 6240
ttcggctcgt cgcgcacgcg ctgggagatg gaggcgaacg gcaccttctc catccagggc 6300
acactcgagc ttgcgtacgt catgggcctc atcaagcact ttgagggtcg cctcaaggac 6360
ggcacgctct acgtcggatg ggtcgacgcc aagacgaacg aaccgctgga cgacaaggac 6420
gtcaaggctg cgtacgagaa gcacattctc gcgcacaccg gcatccgcct catcgagccg 6480
gagatcttca acggctacga cccgaagcgc aagggcttca cgcaggagat cgagatccag 6540
cacgacctcg agcccatcga ggcgtccgag gaggacgcgg ctcgcttcaa gcgcgagcac 6600
ggcgcgctcg tcgacgtcta caccgaggac ggcagcaagt tcttcgtcaa gttcaagaag 6660
ggcgccaagc tgcacattcc caaggctgtt gccttcgacc gccttgtcgc cggacagatc 6720
ccgactggct ggtcgcacaa ggccttcggt atccccgacg acattgcctc gcaggttgac 6780
cgcacctcgc tgtgggcgct cgtctcggtc gccgaggcgc tcatgatggc cggcatcacc 6840
gacccgtatg agctctacaa gtggattcac ccgagcgagg tcggttcgtc gctcggatcc 6900
ggcatgggag gcatcacgag tatctcgaag atgttccgcg accgccgcga ggagaaggac 6960
gtccagaagg acatcctcca ggagaccttc atcaatacgg tcgccggatg ggtcaacctc 7020
ctccttctct cgtcatccgg accgatcaag atccccgtcg gcgcctgcgc gactgccctc 7080
cagtcggtcg agatcgcctg cgacaccatc ctcagcggca aggccaagat catggtctcg 7140
ggaggctacg acgacttctc cgaggagggc tcgtacgagt tcgcaaacat gaaggcgacc 7200
tcgaacagcg agaccgagtt cgctgccggc cgcgagccga acgagatgtc gcgtccgacg 7260
accagcaccc gtgccggctt catggagtcg atgggttgcg gtgctcaggt cctgatgtcg 7320
gcgaagacgg ccatcgagat gggcgccacc atctacggca tcgtcgccta caccgcgacc 7380
gccaccgaca aggctggtcg ctcgattccc gcccccggac gcggtgtcat gggtaccgcg 7440
cgcgagatca cctccaagta cccctcgccc atcctcgatg tcacctaccg ccgccgccag 7500
ctcgagttcc gtcgcaagca gatctcgcag tggctcgaga acgagaccga gctcctcaag 7560
ttcgaggtct cctcgcacgg acaggccaca aagctccccg acgactacgt ctccgagcgc 7620
ctcgcatcca tcgaacgcga agccaagcgc caggaggccg aggctctcgc gacgtacggc 7680
atgctcgccg gccaggaccc gaccatcgcc ccgctccgtc gcgctctcgc cgtttggggt 7740
ctcaccatcg acgacgttgg agtcgcctcg ttccacggca cctcgaccgt tgccaacgac 7800
aagaacgagt cgaacgcgta caacgagcag ttccgtcacc ttggccgcgc caagggtaac 7860
gcctgccccg tcatcgctca gaagtggctc accggacacc cgaagggagg tgccgccgcc 7920
tggatgctca acggcttggc ccaggtcatt cagagcggtc tcgttcccgg caaccgcaac 7980
gccgacaaca tcggcgaaga gcttcgcgcg ttcgagtacc tgctctaccc gtccaagtcg 8040
atccagaccg acggcatcaa ggctggtctc ctcacctcgt tcggcttcgg tcaagtcggt 8100
ggccaggctc tcatcgttca cccgagtctg ctcatcggcg cgctcgagcc cgcccagttc 8160
gaggcgtaca agaagctcaa cgaccagcgc aagaagtggt cataccgtcg cttcaacgat 8220
ttcttcacga acggcaagct cgtcattatc aaggacggca cgcccttcac gcccgagcag 8280
gagaacacga ccctcctcaa cccgctcgtc cgcgccgtgc ccgacaagac tggctcgtac 8340
tcgatgccga aggagttccc tgccaccgtc cctcgcagca acaacgccga agtcgccaac 8400
aagctcgtca gcgcggctgt cggcggtgct ttcggcgtcg gcacggacgt cgagctgatc 8460
agcgccgtcc cgacctcgga gtcgttcctc gagaggaact tcacccagga cgagatcgcc 8520
tactgcaagg ccgcacccga cttccgcgct agcctcgccg cgcgctggtc cgccaaggag 8580
gccactttca aggctctcaa gaccgagtcg aagggcgccg ccgccagcat gcaggacatc 8640
gaggtcgtct ccacgtcgca gggcccgact atcaagctcc acggcgaggt cgagaagatc 8700
gcccaggccg ccggcatcac ggccttcgag gtctcgctct cgcactcgga ggacgtcgct 8760
tgcgccgtcg tcatcgccca gaagtag 8787
<210> 3
<211> 12426
<212> DNA
<213> Aplanochytrium kerguelense
<400> 3
atggaacaac ataaaactga gaaactttca gcagccgatg aaaagttgag tgagagtaca 60
attcattggc ttgctgcaac agatgtccct tcaaactttg tactcttttc cggacaaggg 120
tatcaatact ttgatgagct ccgggaactg tatgaaactg gcacagatga ggttaaaggc 180
ttccttttac ttgcttcgaa aacactgcag gaagagatct gtagtgaaaa agcgaaaact 240
gtcgctttca aatttgtcaa tgagcttgac tttacgaaat ggatagaaga ggatatcgat 300
gtggatcaaa cctacatctt ttctgctcca gtttctttcc ccttaatatt tgtcgctcaa 360
gttgcaaggt acttgcaaac tttaaacttg ctaagaacag accacaaaca atttatccag 420
acaattaaag ggtctagtgg tcatagtcaa gggattgttg ctgctgtatt gatatctaca 480
tcgcctgaca acgacagaat tgttgagaat gcagtcaaac tttgcagata tatgttatgg 540
caagggctga gatgccatga aagcagtgta aacagattca gtggcaaaaa ccacaaatcg 600
aaaaagtcta aagtttattc ctttgatgaa tcgacccgca tggtaaagta ttctcccatg 660
cttgctgtca atggtctcac ggaatccctc ctcaacgatt ttatgaagaa aactgaatcc 720
attattgaag caagaagaac gatacaacaa aagtataacc gaagagaatc attttttgaa 780
acaccaacag aaaaggatga gaaaagtaaa agacttaata tgtttcaaat tgcacttgcg 840
aatggcccca agtcatttgt tgtctctgga gcgccaaaaa cacttattga gctagaacaa 900
agaataaagg agtcaactac atcgtctcca aactctcaaa cacgtatacc atttagcaag 960
cgcaagccag ttgttaaaat gtatttcctc aaggttgggg ctgcttttca tactgaaatt 1020
tgtaatgacg cctctagtaa actaaaagcc gatgtggtcc aaatgaaatt gagtatctcg 1080
ccaaaggaaa tattaattcc tgtctaccat acaaaagatg gttcgaattt atcggagatc 1140
tcagcagatg tgaatgttgt cgatttgcta attgatatgc aaactagtca agtcaatgat 1200
ttcagaaaaa ccttgaagag tatttcaagt cagaatgggg tgtcaaacgt tattgatttt 1260
ggtccaggtg acggaactgc taaattgtgc ctgaaacgaa agtctggaac tggtatacta 1320
gttgtcgcgg ctactggacc actacgaaac agaagatgtg catatggtag aaatcttgtc 1380
ggtctaaact ctgttttgct ggaaaaaaat cctatactag ggatgaattg gggggaagaa 1440
tttaagccca gactgtcgag tcgcaagaat gacaaccaaa tcattgtcgg aactagattt 1500
agtgacttga ctggaaagcc gcctgtaatt ctacctggaa tgacgcctac tacatccttt 1560
cacggcatcg atttggttgc cgcctgtagt aatggtgggt atcatggcga acttgctggt 1620
ggtggattgc cactgccaga atacttcaaa gcgaaaattg atgagctagt gagcaagcaa 1680
aactctggag taggcataaa cataaacatg ctttacctaa actcctactt atggggtttt 1740
caattttcca ctgccattca gatggcgaaa gatggatatc ccatagaatc tatcacttgc 1800
gccgccggag ttccaactac tgaaaaagcg aaagaaataa tgcatcagtt gaaagaagct 1860
ggaattaaat acattgcctt caaacctgga tcaagagccg caattatgga tgtattatct 1920
attgctagag aaaaccccaa tagttcaatc gtactgcaat ggacaggagg acgtggaggt 1980
ggacatcact ccttcgaaga ctttcatgct cctttgctcg acacttatca agaaattaga 2040
gaacacccaa acgttgttct cgtcgtaggc tccggatttg gagatgcaga gaagtcatac 2100
ccatatttag atggttcgtg gtctacattg cctccacata actgtgaatc gagaatgcca 2160
tgtgatgcag ttttatttgg atctaggtgc atggttgcaa aagaagctgc tacctctcca 2220
gaagttaagc aattgattgt taacgctaag ggcgtccgag aaaacgaatc ttgggaaatg 2280
tcttacgaaa acgatgctgg tggaattctc actgtaactt ccgaactcgg cgagcctatc 2340
cataagataa acaaccgcgg gatgaagcta tggagagagt ttgacagaaa gtacttctca 2400
ttaccttcgg gccaagaaag ggaaaaggct atagtgaaag ataaaaagga gataattaga 2460
aggattaatg cagattttca aaaagtttac tttggaagga aggccgatgg aactgttgtg 2520
gacattgcat atatgacata tggtgaaatt ttacggagaa tggtggacct aatgtacgta 2580
accggaggag gagatggaaa acaacataga tttgcgccaa acagatggat agacgttacg 2640
taccaaacta gaacgttcaa gtttttagtt cgcacagaaa gacggttttt gagagacaaa 2700
aaaatggcct ttgcaactga tatgagaaaa ctcgaatctt ttccgataaa gtgcattgat 2760
gagttcgtca agtgctaccc tcaagtagat aaagtgctgg tatcggacga agatgctgga 2820
tattttatcg aactttgtcg aaaccttaga aacggaaaac cggtaaattt tattcctcaa 2880
attgatggct ctttagctta ctggttcaaa aaagactctc tctggtgttc tgaacagatt 2940
gacgcggttc ccgacaatga ccctggcaga gtttgcatac tacatggccc agtagcagcg 3000
caatattcag tagtgtcaga tgagcccata tgcgaaatcc tcggtaacat tcacaaggga 3060
tacgtggaca agttgaaaga ggagaagtat gaggtggcca aagttgaaac aatcggaaag 3120
ctacctgaat acaaaggttc caacaagtct ttccgcaaat ttcttcgtta tggttggttc 3180
aatgccctgt ttcaaagtac cgtaatggtc aaggagaaaa agtgggtcag aaacattatt 3240
ccaggaatca tacaaacgga aaacgaaaac attgtattct cagagcgaac agcaatgaat 3300
gatatgtgtc tcatcgaagt tcttttaaat gattcacaga gaactctagc tttcatagag 3360
tttgaaggaa agcacatttc tgtgtctttg caggattctg aacctgtaaa cagcacactg 3420
accctgtatt ttgagtacca tcctgaaact cctcactctc cgttatttca agtcacaaat 3480
ggaagcaaaa gtattgctag atcaatcaaa cagtattaca ggaatatatg ggattgtcag 3540
gatgttgttt ctatccatga tacattttcc gaaagcttca ccgtcacgag agaagacatc 3600
gaaaggttca atggcgcaat tcaatatgcg gtaaatactt cggagggaac tttagacttt 3660
gcaattgttg caggatggaa atcactgata aaagctctgt tttcaaaaga gatagacggc 3720
agtctgttag atcttgttca catctcccac tcatacaaac tacttgtaga ggagaaggag 3780
agaacgcttg ttcaagctgg agagaaaatt tgttcggaat ctcgcgttat ttcagtacag 3840
atattaccag gtgttggaaa aagtataaca gttaaagggg tactctctag aaacacagtc 3900
aaatgggttg aagtccaatc cgagttttta atccgaggca atttcttcga ttttcaaaag 3960
actttcaagg aatatagcta caaagctgag gttttatgta aagacgatac agttccgcaa 4020
atcctaaact cacaggaatg gctttttctg gactctggag tcagacctct atctaaacac 4080
gataaagttg tatttcaaat cgatcgcgtt gtggaaaaga gaaagtctgt gacaaatatt 4140
tctgatattt tagttgctgg taatgttgtt cgaatcaata cacctgaaga gatcgatggt 4200
tttattgaat tgaaagacca aggaagccca aagggaatca tcattgggaa agttgattta 4260
agccttgaaa atgccaatct gagtgaaaat ccaattatta catatttgga ttccattaaa 4320
gaatcaaaat tgcacggtag tgtttttgaa tcgggaggat atactgtgat gccgaggccc 4380
gaactagtaa cagcgccaag tattgtcgaa agcattgtgt atgcatctgc ttctagagat 4440
atgaatccaa ttcatcggaa tcgcacattc gctaagttag ctggactccc cggtggtagt 4500
accattgtgc acggaatgtg gacagctgca atgagcagaa gaattctaga attacataca 4560
gttctcggag accatcgccg tatttccaca tactcggtta aatttaccgg catggtgttc 4620
ccagaagaca aactagccgt tatggtcaaa cacgttggtg ttacggaagg tagacttatt 4680
cttgatgtag atgttagcaa agtagaaacg caagagcggg ttctccacgg acgagctgaa 4740
gtagaagggc cgtcgacatc attcctattc actggtcaag gatccgcaaa agttgggatg 4800
ggaatggaca gctacgaaga aaagcagtcg gcacgggcag tttggcaaac tgcagataaa 4860
tttttgagga aaaaatttgg attttcaatt gttgatattg tcaaaaataa tccaaaagag 4920
ttgactgttc gtttcggtgg atcacaaggg aaagtgattc gagaaaacta cagagccatt 4980
aaaagggaag atggatctca attgatccga gaaatccata gtaataccac gtcatttaca 5040
tttcgttcac caaatgggct tttatttgcc acgcaatttt ctcagccggc gctggtacta 5100
gtgcaaaaag cagcgtttga agaaatgctt caaggagggt ttgttccgtc aaacagtatg 5160
tttgctggtc attctcttgg tgaatacgca gctcttgcct cgtatgcaaa cgttctgagt 5220
atcgaagact tagttgaaac tgtattttta agaggaatgg taatgcaaaa tgcagttgca 5280
agggacgacg aaggaacatc tgattacggt atggtggctg ccaatccttc aagagtcagt 5340
aaaacgttct cggcacaatt gttgatggaa atcgtccaga aaattgacga cgaatcagat 5400
gacagtcatc ttctccaagt agtaaatttc aatattagag atgctcagta cgtaatagct 5460
ggaaatctgg aattgttaga tagtctatcg aatgttctca atgtaatttc aaacagccac 5520
gacaaaacct ttgacagaag cattatccaa gctgcaatga taaaggctga aaataggaga 5580
agaagctgca aggaaaacaa tcaacctttc cgtcttaagc gtggaatcgc aaccattcct 5640
cttcacggca ttgatgtacc ttttcattct cgacagcttt tgaatggggt acctgctttt 5700
cggtctctac tagaaccaag atttacggaa gaaatgattc aaaagtactt gcctaggttg 5760
atcggaaagt acattcctaa tgtgcacgca gaaccgtttt ccattagcaa agagtacatt 5820
gaaaaggttg caagtgtaac aaaatcgcca gcgttgtaca agcttctatc tacatactca 5880
acaatgtgtg actcatcaaa agcaagaatc ttgctcatag agcttttagc tcatcaattt 5940
gcaatgccag ttcaatggat agatacccag gactatatac tctccaggca cacacagcga 6000
ataattgaga tgggcccagc cccaactctt gttggtatgg cgaaaaagac attgcaatct 6060
agaatatatg gtgacgaaga agactacaaa cctagtattc tgtggtggaa tcaaaaccag 6120
gaggaaatct tctataactt ggacaatcaa ggaatatcta ttcaacagtt tttgcaagaa 6180
ttaaaaggaa ctgaagatga cgagtcacag tctgatgata tgtcatcata tacccagtca 6240
gacgagacta tacaaagaac tgcaaagaac agggttatgc cagaagcaaa tgcaaacgac 6300
aaggagaaag tggggaatga acttcaaaag aaggaagtta caaacaggcc tcttcatgtc 6360
ctaagagtcc tcatctcaac caaacttaag aagaaaatgt tggaggtaaa ggcaacagac 6420
accatcaagg gtgtttctgg tggaaaaagt gcgattcaaa atgagctaat tggagaaata 6480
aacgctgaat acagcagttt tagagatatt gaagatattt cggaagtttc tttagctgag 6540
ctagctagaa gggccccatc ccggcaggaa acagttcttg gtaaagttac aacgaaaatg 6600
gtgaacaaaa tgttttctag caaactgccc ggaaaatttg gtcctaatga tactaaggag 6660
tacttacaaa gcaaactctc ggatgcgtcc acagtcgatt cagtgtctct tcatgccctg 6720
actctgcagc cggtttctag actagctacc cctggagagg cacaggcatg gcttgattct 6780
gtttgtagag actttgaaaa cgatcagaga atcagtattc taaagaacaa aagtccaaat 6840
tcgaacgaaa gtgtgatgat ttctgaacaa aggactgttc aaaacaactt tgtacaaatt 6900
tctgaagaac cagtaaacag caaacatgta ttgttgtccc tgatatctat gaagctacag 6960
aaggaaatct cctcaatcaa tgaaacatca acaataaaga gcttggtgaa tgggaagagt 7020
gcagttcaaa acgaaattct tggagaaatt gcttccgagt ttcggacaga cgaatctgaa 7080
ggagctgcag atatttcgct ctccgagctt ggagaaaagc ttggaattgg gtatgcagga 7140
cctggcagtg ttgtttccag gcacataaat aaaatgttaa actccaagct tccagggtca 7200
tttaattctt catcagccaa gaagtatctt tcgatgcaag gctttggaaa aggacgaagt 7260
gactccgtac tggttcatgc tttgacaatg caaccaaaat cgcgcctgca gggagtcgaa 7320
gatgcaaagt cttggttgga ttcggtttgt tcgagttatg ggaaatgggc aaatattgat 7380
ctcacagttt ctcgtactct ctcttcaggt gatagtagtc ctgctcattc gacaaaagat 7440
atcctttcgt ccactgcaat gaagaaattg aaaaaattgt acttggacca ggtggatgtc 7500
ttcaacactt ttttgggaga ggatcgacgg gccaaacttc aaaaggttct aacagaggaa 7560
accagtgcaa aagatagaat agaggctcag ctccttgaat taaaggaaga acttggagac 7620
cctttttgtg acgggataca accccagttt aatgaagctc ggatacgaat ctatgattca 7680
tactggaact gggtggtgca ggatgcatta gaactccatt atcatacatt ctcttgtgtg 7740
ttgaactcaa aaggcaagaa cgtaaatatt cccaattcat ctaacagcta ttttcgcgca 7800
atgagcgact ggataacttc ttcaaacgaa aagttggaag aaaataaacc tccgcaggct 7860
tggtttcgca attacctttg taatcgtgcc actccagaac ttctaaccgt tgtacaatac 7920
tttgcagcaa gaatgaacga acaaggacat tcagaatatg ctcaagcagt atctctttta 7980
gcagaacaag tatcacagtg gatatctaga ccacctgtac acatagcatt gtttgcatct 8040
ctagaaccta gggtgacagt tgatagtgct aacaattttg gattgaaata tgaagaaaaa 8100
ccaagaaagt ttgtggctgt cggtgaaaac aaagctttct cgtgtgataa tgcgtccctg 8160
tatgttaaag aaatgagcaa ggggctattt tatgaccacc gtgtggcttc gaaagttgaa 8220
catcccagcc aaagtgtata cctttctccc gataattctt tctttccaaa cgaggaagag 8280
agtgaccaga tttcaagcgg aatgagacta ccaaaaacac aaggagaaat ccagcgtgat 8340
ttttcaagac tccctgctgg tcagaagcta gaagtcatga gaaagagtgt taatcggtcg 8400
tccgacacag atgaggaaat tgccaacatc gttcgacaaa actataacag tatccatgtg 8460
gcaaaaaacg tacctttggt gcatttgaaa tcgccttcta aatttgacaa gacggtaaga 8520
gtactcgatg agcctttgac atcaatgtac ttgagttgtt tgcatgatat tgcaacaagt 8580
ggagttagct ttgcaggaca aaacgctttg gtgacaggag ccggctttgg ttctattggg 8640
attgaattaa tcaaaccgtt gcttgagggg ggggcaacgg ttttagtaac agttagactt 8700
aatcggactg acgagcaaat gcaaattgtt aatgaaaggt ttcaaagatt gtacgaagag 8760
tttgggtcca gaggaagcaa gctcgttctg gttccttgca attgcgcttc aaatcaagat 8820
gtacattccc tcattagaca catctatgag aaattgaaac ttgacctaga ttttatattt 8880
ccatttgctg ctatcgggga gcaagggaaa gatgtgtctg aaattggctc caaatctgaa 8940
gtcgctcata gactaatgct gactaatact ataagattac taggtgcagt gaaaaaggcc 9000
aaagaggata gatgcattga aacaaggcct gcactggttc tgttgccatg ctctccaaac 9060
catggggatt ttggactaga tgggttatat gcagaatcaa agcttggatt ggaatcccta 9120
gtaaacaagt ggaaaagtga agaatgggga aattacctaa gtatttgcgc tgcagtcata 9180
ggttggacaa ggtccaaact catgtggcag aacaatgtcg ttgccgaggg aatagaaaag 9240
ttgggagtga gaaccttttc tactacagag actgctttta acctgattgg tctactacat 9300
cctgatatcg tgtctcatgc agcagaagaa cctctatggg cagaccttac tggaaactgg 9360
ggcgctgctc ctgacttgaa ggaacattca aaaaggattc gaacgtcact tctggcacaa 9420
agtaaagcag caaaagccat atcgctgtcg tcaaaatcgc tcgttataga accttctgaa 9480
tccaagacga aagcgcagac agaaattgtt aacacatacc tagctggtgt gaatgataag 9540
ttgaggcttc cgctagcaaa tcccgaaaag ttctgcaatc cattccctaa aattccatcc 9600
caggaacgaa tggattcatt agcatacctt aaacactcag tagatctcaa aaaagtcgta 9660
gtagtagtag gatatggaga aattgggcct tggggaaatt cgcgcactag atgggagatg 9720
gagtcattcg gtgaattctc gcttgaaggt gcaatcgaac ttgcttggct cgttggctta 9780
atcaaaccag ttactggacc gttaaaaaat gatcctagaa cgcaatactt tggttgggta 9840
gatgcagaat cagaagagcc agttgcagat catgaaataa agactcgata tgagaaagta 9900
ctgcttcagc actctgggat tcgtcttatt gaaccggagc tctttgaagg atataatcca 9960
aagaaaaagt caattctgag acaagtagcc atcgctgaag acatgaaacc aatagaagta 10020
gcttctttgg aagaagccca gcagtacgtg aatgagttag ggaaagagtt catagacgtt 10080
tttaatgaaa acgttgaatc taatgatgga caatggtata ttcgcctaaa agcaggagca 10140
gtggtctcca ttcctggagc gttaagcttc aacagattta ttgctggtca gctacctaca 10200
ggatgggatg caaaacgact aggaatacca gatgatattg ccgactctgt agatcctgtt 10260
acgctatatg ctctggtttc aacagttgaa gccttggtgt gtgctggatt gacagatcca 10320
tatgaactgt atcaatatgt gcatgtaagt caagttggaa acacgtcagg tggcggcatg 10380
ggagggatgc gatcattgaa gcgaatgttc ctagaaagaa aacttgatgc agaaatccct 10440
tctgatactt tagcagagtc atttattaac acaatgccag cctgggtgaa catgctgctt 10500
ctgtcaagct cgggaccaat aaagactcca gtcggtgctt gcgccacagc ggcagagtcg 10560
gtagacatag gaattgaaac catcttagca ggaaaagcca gagtcgttat agcgggcggg 10620
tatgatgact tttgtgaaac gggaagtaat gagtttgcca tgatgggagc aacttccaac 10680
agtcaaacag aagctgaaaa aggcagattt ccacgtgagg catcaagacc aatgacagat 10740
acacgtgccg gattcatgga atcacaaggc gctggtatgc aagtgttaat ggatgctgaa 10800
ttagccattc agatggggct tccagtatat ggtattcttg ctctgtcaaa cactgctacc 10860
gatagacaag gaagatccgt gcccgcccca ggtcgtggaa tactaaccac tgcgagagaa 10920
gtgcgctcaa aaaactctaa aagtagccac aacggcaaac cagtagaaaa tcctctattg 10980
tctgtaaagt tccgcaaaaa acatttgaga caagagctcg atgctataga tctgtgggcc 11040
agccaagaga tatcgaacat gtctgaggaa ttctcacaga attcaagaaa aatggaaaac 11100
aggaaaggtt ttgtagaaac aatgcgaaaa aagaagcgtt cggcagcttt tgaaacctgg 11160
ggacaaggat tctataggaa tgatgactcc atagcccctt tacgcggagc tttgtccgta 11220
tggggattaa cagttgatga tcttcttgtg ggaagttttc acggaactgg aaccaatctt 11280
aacgatacga atgagtcctc cttagtgaat aagcagctta agcatttgga tcgaaaagaa 11340
ggaaacattc tgttagtagt aactcaaaaa tatctgactg gtcatccaaa gggtgcagct 11400
gctgcctgga tgctcaatgg attgctgcaa tgcatgaatt ctggaagagt accgggaaac 11460
cgaaacttgg acaatgtaga tggaaagctc agaacgaacg gttacttgtt ctatccaaat 11520
cggacaattg aggttccaag agtggaagca gcttttctta aaagctttgg atttggtcaa 11580
gccggagctg aagttgtcat tattcatcct gatcgtctcc tggctgtttt aagtgaagaa 11640
aatctgaaat cgtatatact tcgaagaaat gaaagagaga aaagagcata tcgatatcat 11700
caaggcgtaa tgagtggaca tcatactatg gtgcaagtga aggaatttgc tccatatgaa 11760
gacgatatcc tagaagaaat atatttgaat ccaagagcta gggcttcctt tgactcatcg 11820
aaaagtactt ggacgtttca caaatatagg aacgacaaag atactgtaga cgataaagat 11880
gaaatggagc aagattcatt gaatgaatca gtagatcttc cgaaagatat tacaaaagtt 11940
ggatcaccct ctgaaatacg tattccagta aaaacaagat tggaagttac tgttcgtgaa 12000
ggagtcgaag gtttgacgaa aaaggacaag ttttcaagtc aaggggttgg cgttgatgtt 12060
gaaccagttt caacttttgc acaacatgaa gagaaaacga tctttattca gaacaacttc 12120
actgagaatg aacagctgta ttgtaatcat gctgcaagtc cagcagcaag ctatgcgggt 12180
agatgggcag cgaaggaagc agtgattaaa gccatcagca attcttcact tgaaacaaga 12240
tcactttggc aaggagctga gggtaaactg atcgacattg aaatcattca aagtaattct 12300
ggtgctcctg aggtagtctt gcatggtcat gcaaaagaag tattccaaac tttaggtctt 12360
acaaatgtta aggtttctat cagtcatact ccagaagttg cagtagccca agcaataaca 12420
aattaa 12426
<210> 4
<211> 5664
<212> DNA
<213> 酿酒酵母
<400> 4
atgaagccgg aagttgagca agaattagct catattttgc taactgaatt gttagcttat 60
caatttgcct ctcctgtgag atggattgaa actcaagatg tttttttgaa ggattttaac 120
actgaaaggg ttgttgaaat cggtccttct ccaactttgg ctgggatggc tcaaagaacc 180
ttgaagaata aatacgaatc ttacgatgct gctctgtctt tacatagaga aatcttatgc 240
tattcgaagg atgccaaaga gatttattat accccagatc catccgaact agctgcaaag 300
gaagagcccg ctaaggaaga agctcctgct ccaactccag ctgctagtgc tcctgctcct 360
gcagcagcag ccccagctcc cgtcgcggca gcagccccag ctgcagcagc tgctgagatt 420
gccgatgaac ctgtcaaggc ttccctattg ttgcacgttt tggttgctca caagttgaag 480
aagtcgttag attccattcc aatgtccaag acaatcaaag acttggtcgg tggtaaatct 540
acagtccaaa atgaaatttt gggtgattta ggtaaagaat ttggtactac tcctgaaaaa 600
ccagaagaaa ctccattaga agaattggca gaaactttcc aagatacctt ctctggagca 660
ttgggtaagc aatcttcctc gttattatca agattaatct catctaagat gcctggtggg 720
tttactatta ctgtcgctag aaaatactta caaactcgct ggggactacc atctggtaga 780
caagatggtg tccttttggt agctttatct aacgagcctg ctgctcgtct aggttctgaa 840
gctgatgcca aggctttctt ggactccatg gctcaaaaat acgcttccat tgttggtgtt 900
gacttatcat cagctgctag cgctagtggt gctgccggtg caggtgctgc tgccggtgca 960
gctatgatcg atgctggcgc tctggaagaa ataaccaaag accacaaggt tttggcgcgt 1020
caacaactgc aagtattggc tcgttatcta aaaatggact tggataacgg tgaaagaaag 1080
ttcttgaaag aaaaggacac tgttgctgaa cttcaagctc agttggatta cttgaatgcc 1140
gaattaggtg aattctttgt taacggtgtt gctacttctt tctctagaaa aaaggccaga 1200
accttcgatt cttcctggaa ctgggctaaa caatctttat tatcattata ctttgagata 1260
attcatggtg tcttgaaaaa cgttgataga gaggttgtta gtgaagctat caatatcatg 1320
aacagatcta acgatgcttt gattaaattc atggaatacc atatctctaa cactgatgaa 1380
acaaaaggtg aaaactatca attggttaaa actcttggtg agcagttgat tgaaaactgt 1440
aaacaagttt tggatgttga tccagtttac aaagatgttg ctaagcctac cggtccaaaa 1500
actgctattg acaagaacgg taacattaca tactcagaag agccaagaga aaaggttagg 1560
aaattatctc aatacgtaca agaaatggcc cttggtggtc caatcaccaa agaatctcaa 1620
cctactattg aagaggattt gactcgtgtt tacaaggcaa tcagtgctca agctgataaa 1680
caagatattt ccagctccac cagggttgaa tttgaaaaac tatatagtga tttgatgaag 1740
ttcttggaaa gctccaaaga aatcgatcct tctcaaacaa cccaattggc cggtatggat 1800
gttgaggatg ctttggacaa agattccacc aaagaagttg cttctttgcc aaacaaatct 1860
accatttcta agacggtatc ttcaactatt ccaagagaaa ctattccgtt cttacatttg 1920
agaaagaaga ctcctgccgg agattggaaa tatgaccgcc aattgtcttc tcttttctta 1980
gatggtttag aaaaggctgc cttcaacggt gtcaccttca aggacaaata cgtcttgatc 2040
actggtgctg gtaagggttc tattggtgct gaagtcttgc aaggtttgtt acaaggtggt 2100
gctaaggttg ttgttaccac ctctcgtttc tctaagcaag ttacagacta ctaccaatcc 2160
atttacgcca aatatggtgc taagggttct actttgattg ttgttccatt caaccaaggt 2220
tctaagcaag acgttgaagc tttgattgaa tttatctacg acactgaaaa gaatggtggt 2280
ttaggttggg atctagatgc tattattcca ttcgcggcca ttccagaaca aggtattgaa 2340
ttagaacata ttgattctaa gtctgaattt gctcatagaa tcatgttgac caatatctta 2400
agaatgatgg gttgtgtcaa gaagcaaaaa tctgcaagag gtattgaaac aagaccagct 2460
caagtcattc taccaatgtc tccaaaccat ggtactttcg gtggtgatgg tatgtattca 2520
gaatccaagt tgtctttgga aactttgttc aacagatggc actctgaatc ctgggccaat 2580
caattaaccg tttgcggtgc tattattggt tggactagag gtactggttt aatgagcgct 2640
aataacatca ttgctgaagg cattgaaaag atgggtgttc gtactttctc tcaaaaggaa 2700
atggctttca acttattggg tctattgact ccagaagtcg tagaattgtg ccaaaaatca 2760
cctgttatgg ctgacttgaa tggtggtttg caatttgttc ctgaattgaa ggaattcact 2820
gctaaattgc gtaaagagtt ggttgaaact tctgaagtta gaaaggcagt ttccatcgaa 2880
actgctttgg agcataaggt tgtcaatggc aatagcgctg atgctgcata tgctcaagtc 2940
gaaattcaac caagagctaa cattcaactg gacttcccag aattgaaacc atacaaacag 3000
gttaaacaaa ttgctcccgc tgagcttgaa ggtttgttgg atttggaaag agttattgta 3060
gttaccggtt ttgctgaagt cggcccatgg ggttcggcca gaacaagatg ggaaatggaa 3120
gcttttggtg aattttcgtt ggaaggttgc gttgaaatgg cctggattat gggcttcatt 3180
tcataccata acggtaattt gaagggtcgt ccatacactg gttgggttga ttccaaaaca 3240
aaagaaccag ttgatgacaa ggacgttaag gccaagtatg aaacatcaat cctagaacac 3300
agtggtatca gattgatcga accagagtta ttcaatggtt acaacccaga aaagaaggaa 3360
atgattcaag aagtcattgt cgaagaagac ttggaaccat ttgaggcttc gaaggaaact 3420
gccgaacaat ttaaacacca acatggtgac aaagtggata tcttcgaaat cccagaaaca 3480
ggagagtact ctgttaagtt actaaagggt gccactttat acattccaaa ggctttgaga 3540
tttgaccgtt tggttgcagg tcaaattcca actggttgga atgctaagac ttatggtatc 3600
tctgatgata tcatttctca ggttgaccca atcacattat tcgttttggt ctctgttgtg 3660
gaagcattta ttgcatctgg tatcaccgac ccatacgaaa tgtacaaata cgtacatgtt 3720
tctgaggttg gtaactgttc tggttctggt atgggtggtg tttctgcctt acgtggtatg 3780
tttaaggacc gtttcaagga tgagcctgtc caaaatgata ttttacaaga atcatttatc 3840
aacaccatgt ccgcttgggt taatatgttg ttgatttcct catctggtcc aatcaagaca 3900
cctgttggtg cctgtgccac atccgtggaa tctgttgaca ttggtgtaga aaccatcttg 3960
tctggtaagg ctagaatctg tattgtcggt ggttacgatg atttccaaga agaaggctcc 4020
tttgagttcg gtaacatgaa ggccacttcc aacactttgg aagaatttga acatggtcgt 4080
accccagcgg aaatgtccag acctgccacc actacccgta acggttttat ggaagctcaa 4140
ggtgctggta ttcaaatcat catgcaagct gatttagctt tgaagatggg tgtgccaatt 4200
tacggtattg ttgccatggc tgctaccgcc accgataaga ttggtagatc tgtgccagct 4260
ccaggtaagg gtattttaac cactgctcgt gaacaccact ccagtgttaa gtatgcttca 4320
ccaaacttga acatgaagta cagaaagcgc caattggtta ctcgtgaagc tcagattaaa 4380
gattgggtag aaaacgaatt ggaagctttg aagttggagg ccgaagaaat tccaagcgaa 4440
gaccaaaacg agttcttact tgaacgtacc agagaaatcc acaacgaagc tgaaagtcaa 4500
ttgagagctg cacaacaaca atggggtaac gacttctaca agagggaccc acgtattgct 4560
ccattgagag gagcactggc tacttacggt ttaactattg atgacttggg tgtcgcttca 4620
ttccacggta catccacaaa ggctaatgac aagaacgaat ctgccacaat taatgaaatg 4680
atgaagcatt tgggtagatc tgaaggtaat cccgtcattg gtgttttcca aaagttcttg 4740
actggtcatc caaagggtgc tgctggtgca tggatgatga atggtgcttt gcaaattcta 4800
aacagtggta ttattccagg taaccgtaac gctgataacg tggataagat cttggagcaa 4860
tttgaatacg tcttgtaccc atccaagact ttaaagaccg acggtgtcag agccgtgtcc 4920
atcacttctt tcggttttgg tcaaaagggt ggtcaagcta ttgtggttca tccagactac 4980
ttatacggtg ctatcactga agacagatac aacgagtatg tcgccaaggt tagtgccaga 5040
gagaaaagtg cctacaaatt cttccataat ggtatgatct acaacaagtt gttcgtaagt 5100
aaagagcatg ctccatacac tgatgaattg gaagaggatg tttacttgga cccattagcc 5160
cgtgtatcta aggataagaa atcaggctcc ttgactttca actctaaaaa catccaaagc 5220
aaggacagtt acatcaatgc taacaccatt gaaactgcca agatgattga aaacatgacc 5280
aaggagaaag tctctaacgg tggcgtcggt gtagatgttg aattaatcac tagcatcaac 5340
gttgaaaatg atacttttat cgagcgcaat ttcaccccgc aagaaataga gtactgcagc 5400
gcgcagccta gtgtgcaaag ctctttcgct gggacatggt ccgccaaaga ggctgttttc 5460
aagtccttag gcgtcaagtc cttaggcggt ggtgctgcat tgaaagacat cgaaatcgta 5520
cgcgttaaca aaaacgctcc agccgttgaa ctgcacggta acgccaaaaa ggctgccgaa 5580
gaagctggtg ttaccgatgt gaaggtatct atttctcacg atgacctcca agctgtcgcg 5640
gtcgccgttt ctactaagaa atag 5664
<210> 5
<211> 8835
<212> DNA
<213> 圆红冬孢酵母
<400> 5
atggtcgcgg cgcaggactt gccgctcgcg ctgagcatca gcttcgcgcc cgagtcgtcg 60
accatctcga tgacgctgtt caaccagccc gaggcgtcga aacccgccct ccccctcgag 120
ctcaagtaca agtacgaccc ctcgacgccg tacgccccga tccacgagat caccgaggac 180
cgtaatcaga ggatcaagca gcactactgg gacctctggg gcctcggcaa caaggcagac 240
cagggcatct cgcagctcaa gatcaccgac gagttccagg gcgacctcgt caccatctcg 300
gccgacgaga tcgaggcgtt ctgccgtgtt gtcggcatcg agggcgaggc gtacaagcgc 360
aaccacaagg ccggcatgca ggtcccgctc gacttcgcca tcaagctcgg ctggaaggcc 420
atcatgaagc cgatcttccc ctcgacgatt gacggcgacc tgctcaagct cgtccacctc 480
tcgaacggct tccgcgtcct ccccgacacg cccacactcc aggttggcga cgtcgtgacg 540
accacgtcgc gcatcgaatc aatcacgaac tcggacacgg gcaaaaccgt ctcggttcgc 600
ggcgtcatct cgctcgtctc gtccgccgac tcgaagggca aggacgcctc gaccgaggac 660
cgcatcccgc tcatcgaggt cacctcgtcc ttcttctacc gcggcaagtt cagcgactac 720
gcccagacat tctcccgcgt cgcccacccg acctactctg tcccgatcac cacgcccgag 780
gccgtcgccg tcctccagtc caaggagtgg ttccagtggg acgacgactc gaagcccctc 840
gaggtcggca ccaagctcca gttcaaggtc gagtcgaact atgtctacgc cgacaagtcg 900
tcctacgcga tggctaccgt caccggcggc gcgtacgtca tcacccccga gctcaagctc 960
gctgtcaagg ttgccacggt cgactacacg tccgagggcg agggcgtcat ccagggcgac 1020
ccggtcatcg agtacctcaa gcgccacggc tcggccctcg accagcccat catgctcgag 1080
aacggcggct attcgctcac caaggccggc cagtgcacct tcacgacgcc cgcgtccaac 1140
ctcgactact cgctcacctc gggcgacacg aacccgattc acacgaaccc gtactttgcc 1200
tcgctcgcct acctccccgg caccatcacg cacggcatgc actcgtcggc ccgcacgcgc 1260
aagtttgtcg agcaggtcgc cgcagacaac gtcggcgcgc gcgtccgcaa gtacgaggtc 1320
ggcttcacgg ccatgtgcct cccctcgcgc aagatggagg tccgccttaa gcacgtcggc 1380
atgaccgcgg acggaaaccg cctcatcaag gtcgagaccg tcgacgtcga gggcggcaac 1440
gtcgttctca gcggaaccgc cgaggtcgcc caggctccca ccgcgtacgt cttcaccggt 1500
caaggttcgc aagagcccgg catgggcatg gagctctacg ccaactcgcc cgtcgcccgc 1560
gccgtctggg acgaggctga ccgccacctc ggcgaggtct acggcttctc catcctcgag 1620
attgtccgta cgaaccccaa ggaaaagact gtgcacttcg gcgggttgaa aggccaagca 1680
acccgtcaga agtacatgga catgtcgtac acaacgactg accatgaggg caacgttaag 1740
actctcccgc tcttcggcga catcgacctc cgtacctcac gctacacgtt ctcgtcgccg 1800
accggtctcc tctacgccac ccagttcgcc cagatcgccc tcgtcgtaac ggagaaggcc 1860
gccttcgagg acatgcgcgc caagggtctc gttcagaagg actgcgtctt tgccggtcac 1920
tcgctcggag agtactcggc tctcgcctcg atcgccgaca tcctccccat ctcggccctc 1980
gtcgacgtcg tcttctaccg cggtatcacc atgcagcgcg ccgtcgaacg cgaccacctc 2040
aaccgctcgt cgtacggaat ggtcgccgtc aacccgagcc gcatcggcaa gagctttggc 2100
gacgccgccc tccgcgaggt cgtcgacacc atcgcccgcc gcggaaacat cctcatcgag 2160
gtcgtcaact acaacgtcga gggacagcaa tacgtcgtcg ccggtcacct cgtcgccctc 2220
caatccctca caaacgtcct caacttcctc aagatccaga agatcgacct cgccaagctc 2280
accgagacga tgtcgatcga gcaggtcaag gagcacctgt gcgagatcgt cgacgagtgc 2340
gtccagaagg cgcgcgacct ccaggccaag acgggcttca tcaccctcga gcgcggcttt 2400
gcgacgatcc cgctccccgg tatcgacgtg ccgttccact cgcgctacct ctgggcggga 2460
gtcatgccgt tccgcactta cctctcgaag aaggtcaacc cggcgcactt caacgccgac 2520
ctcctcgtcg gccgctacat ccccaacttg accgccgtcc actacgaggt ctcgaaggag 2580
tacgccgaac gcatccacac ccagacgtcg tcgccgcgcc tcaacaagat tctcaaggcc 2640
tgggacgagg agcgctgggg cgcacccgag aaccgcaaca agctcggcta cgccatcctc 2700
atcgagctcc tcgcgtacca gttcgcctcg cccgtccgct ggatcgagac gcaggacatc 2760
ctcttccgcg acttcaagtt tgagcgcctc gtcgagcttg gcccgtcgcc cactctcacc 2820
ggcatggcta cgcgcacgca gaagctcaag tacgacgcgc acgactcgtc ggtcggcatc 2880
aagcgctcga tctactgcat cgccaagcac cagaaggaga tctactacca gttcgatgac 2940
gttgccggcg aagaggcgcc cgctcctgcc gcagttgcgc cttccgctcc cgctcccaag 3000
gccgccccag tcgccgccgc ccctccccct cccgctcctg tcgctgccgc gcctgccgcc 3060
gccgtcgccg acgagccgct caaggctgtc gacacgctcc gcatcatcat cgcgcagaag 3120
ctcaagaagc ccgttggcga agtccccctc accaagtcga tcaaggagct cgtcggcggc 3180
aagtcgaccc tccagaacga gattctcggc gaccttcaag gcgagttcag cagcgcgcct 3240
gaaaagggcg aggagatgcc tctccaggag ctcggcgcgg ccctccagca gggctactct 3300
ggcaagctcg gcaagtacac caccggcgtc atctcgcgca tgattggcgc caagatgccc 3360
ggcggttttg gtctctccgc cgtccagggt cacctcggca agacctacgg cctcggcgcc 3420
ggtcgcatcg atggcgtcct cctcttcgcc gtcacgcagg agccggctaa gcgtctcgcc 3480
aacgagggtg aggcgaaggc ttgggtcgac tcggtcgcgc aaggctacgc ctcgatggct 3540
ggcatctcgc tcgccgccgg cggtggagct gctgctgctg cccccgcgat ggcgttcgcc 3600
gctccggccg cagctggcgg tggagcgccc gctaagacta tattgatatt gggtgactca 3660
ttgtccgctg gttatggtat taatcctgaa caaggttggg tcgccttatt gcaaaagaga 3720
ttggatcaac aattcccaaa gcaacataaa gtaatcaatg catctgtttc aggtgaaact 3780
acatctggtg ctttggcaag attaccaaag ttgttaacca cttacagacc taacgttgtc 3840
gtaattgaat tgggtggtaa cgacgcctta agaggtcaac cacctcaaat gatccaatca 3900
aatttggaaa agttaataca acactcccaa aaagctaaga gtaaggttgt cgtattcggt 3960
atgaagatcc cacctaacta tggtacagca tactctcaag ccttcgaaaa taactataag 4020
gttgtctcac aaacctacca agtcaaattg ttaccatttt tcttggatgg tgttgctggt 4080
cataagtcct taatgcaaaa tgaccaaatc cacccaaacg ccaaagctca aagtatattg 4140
ttgaacaacg cttaccctta catcaagggt gcattagctg gtggaggcgg aggcggcgcg 4200
gctgtcggcg gcgccggctt tatgatcaac accgagcagc tcgacaagat gcaggagaag 4260
caggacaact tcgtctcgca gcaggtcgag ctcttcctcc gctacctcgg caaggactcg 4320
cgcgagggcc accgcctcgc cgacatgcag aaggcagagg tcgccaacct ccaggagaag 4380
ctcgactcga tcgctcgcga gcacggcgac gcctatgtcc agggcatcca gcccgtcttc 4440
gacccgctca aggcccgcca cttcaactcg tcgtggaact gggtccgtca ggacgcgctc 4500
atgatgtgga tggacatcct cttcggccgc ctcaccaccg tcgaccgcga catcaccgct 4560
cgctgccttg tcatcatgaa ccgcgccgac ccttctctca tcgactacat gcagtacacc 4620
atcgacaaca cccccgtcga gcgcggcgag cattacgtcc tcgccaagca attcggccag 4680
cagctcctcg acaactgccg cgagatgatc ggccaggctc cgctctacaa ggacgtcacc 4740
ttcccgaccg cgcccaagac gaccgtcaac gccaagggcg acatcatcac cgaggaggtc 4800
aaccgccccg gcgtctctcg cctcgagaag tatgtcgccg agatggctgc cggctcaaag 4860
gtcaccgtcg ccagcgtcaa cctcgacaag gtccaggagc aggtcgagaa gctgtacaag 4920
ctcgtcaagt cgcagccgca gatttcgaag cagcacatga cgtcgatcaa gtcgctgtac 4980
gctgaggtcg ttcgcggtct cggcaaggac gccggccctc ctccggtcca caaggccggc 5040
actcgcgccc gccgcccctc gagccagttc ctccgtcccg cagccgtctc cgaggcgact 5100
ttcctccccg aggacaaggt gcctctcctg cacctcaagc gcaagatcgg caacgactgg 5160
caatactcga gcaagctcac gtcgctctac ctcgacatcc tcaaggagat tgccacgtcg 5220
ggtgtcacct tcgagcacaa gaacgcgctc atgaccggtg tcggcaaggg ctccatcggt 5280
atcgagatcg tcaagggtct cctcgctggt ggcgctcgcg tcgtcatcac gacctcgcgc 5340
tactcgcgct cgactgtcga gtactaccag gcgatctacc aggaggtcgg ctcgaagggc 5400
tcgtcgctca ccgtcgtccc cttcaaccag ggctcgaagc aggatgtcga ggcgctcgtc 5460
gacttcattt attcgaagga taagggtctc ggcatggacc tcgactacat cctccccttc 5520
gccgcccttc ccgagaacgg ccgcgagatc gacggcatcg acgaccgctc cgagctcgcc 5580
caccgcatca tgctcaccaa cctcctccgc ctcctcggtg ccgtcaagtc gaagaaggcc 5640
gccctcaagc tcacgacccg cccaaccgag gtcgtcctcc cgctttcgcc gaaccacggc 5700
ctcttcggca acgacggtct ctactcggag tcgaagatct cgctcgagac gctcttcaac 5760
cgctggagct cggagagctg gggcgagtac ctctgcctcg ctggcgctgt catcggatgg 5820
acgcgcggta ccggtctcat gtcggcgacg aactcggtcg ccgaaggtat cgaggcgcag 5880
ggttgcagga cgttctccgc caaggagatg gccttcaaca ttctcggcct catgcacccg 5940
ctcgtcttcg acgtcgcgca gatcgagcct gtctgggccg acctcaacgg tggcatggac 6000
aagctccccg accttgccaa cctcacgacc gagatccgca agaagctcaa cctcaccgcg 6060
tcgacccgcc gcgccatcgc caaggacaac tcgttcgact acaaggtcgc gcacggcccg 6120
gcgatggagc agatacacca gcggatcaac gtcgccccgc gcgccaactt ctcccttccc 6180
ttccccgagc tcaagccgat cgatgccaag tcggagctcg cgaagctccg tggcctcatc 6240
gacctcgaga aggtcgtagt catgaccggt tacgccgagg tcggaccgtt cggctcgtcg 6300
cgcacgcgct gggagatgga ggcgaacggc accttctcca tccagggcac actcgagctt 6360
gcgtacgtca tgggcctcat caagcacttt gagggtcgcc tcaaggacgg cacgctctac 6420
gtcggatggg tcgacgccaa gacgaacgaa ccgctggacg acaaggacgt caaggctgcg 6480
tacgagaagc acattctcgc gcacaccggc atccgcctca tcgagccgga gatcttcaac 6540
ggctacgacc cgaagcgcaa gggcttcacg caggagatcg agatccagca cgacctcgag 6600
cccatcgagg cgtccgagga ggacgcggct cgcttcaagc gcgagcacgg cgcgctcgtc 6660
gacgtctaca ccgaggacgg cagcaagttc ttcgtcaagt tcaagaaggg cgccaagctg 6720
cacattccca aggctgttgc cttcgaccgc cttgtcgccg gacagatccc gactggctgg 6780
tcgcacaagg ccttcggtat ccccgacgac attgcctcgc aggttgaccg cacctcgctg 6840
tgggcgctcg tctcggtcgc cgaggcgctc atgatggccg gcatcaccga cccgtatgag 6900
ctctacaagt ggattcaccc gagcgaggtc ggttcgtcgc tcggatccgg catgggaggc 6960
atcacgagta tctcgaagat gttccgcgac cgccgcgagg agaaggacgt ccagaaggac 7020
atcctccagg agaccttcat caatacggtc gccggatggg tcaacctcct ccttctctcg 7080
tcatccggac cgatcaagat ccccgtcggc gcctgcgcga ctgccctcca gtcggtcgag 7140
atcgcctgcg acaccatcct cagcggcaag gccaagatca tggtctcggg aggctacgac 7200
gacttctccg aggagggctc gtacgagttc gcaaacatga aggcgacctc gaacagcgag 7260
accgagttcg ctgccggccg cgagccgaac gagatgtcgc gtccgacgac cagcacccgt 7320
gccggcttca tggagtcgat gggttgcggt gctcaggtcc tgatgtcggc gaagacggcc 7380
atcgagatgg gcgccaccat ctacggcatc gtcgcctaca ccgcgaccgc caccgacaag 7440
gctggtcgct cgattcccgc ccccggacgc ggtgtcatgg gtaccgcgcg cgagatcacc 7500
tccaagtacc cctcgcccat cctcgatgtc acctaccgcc gccgccagct cgagttccgt 7560
cgcaagcaga tctcgcagtg gctcgagaac gagaccgagc tcctcaagtt cgaggtctcc 7620
tcgcacggac aggccacaaa gctccccgac gactacgtct ccgagcgcct cgcatccatc 7680
gaacgcgaag ccaagcgcca ggaggccgag gctctcgcga cgtacggcat gctcgccggc 7740
caggacccga ccatcgcccc gctccgtcgc gctctcgccg tttggggtct caccatcgac 7800
gacgttggag tcgcctcgtt ccacggcacc tcgaccgttg ccaacgacaa gaacgagtcg 7860
aacgcgtaca acgagcagtt ccgtcacctt ggccgcgcca agggtaacgc ctgccccgtc 7920
atcgctcaga agtggctcac cggacacccg aagggaggtg ccgccgcctg gatgctcaac 7980
ggcttggccc aggtcattca gagcggtctc gttcccggca accgcaacgc cgacaacatc 8040
ggcgaagagc ttcgcgcgtt cgagtacctg ctctacccgt ccaagtcgat ccagaccgac 8100
ggcatcaagg ctggtctcct cacctcgttc ggcttcggtc aagtcggtgg ccaggctctc 8160
atcgttcacc cgagtctgct catcggcgcg ctcgagcccg cccagttcga ggcgtacaag 8220
aagctcaacg accagcgcaa gaagtggtca taccgtcgct tcaacgattt cttcacgaac 8280
ggcaagctcg tcattatcaa ggacggcacg cccttcacgc ccgagcagga gaacacgacc 8340
ctcctcaacc cgctcgtccg cgccgtgccc gacaagactg gctcgtactc gatgccgaag 8400
gagttccctg ccaccgtccc tcgcagcaac aacgccgaag tcgccaacaa gctcgtcagc 8460
gcggctgtcg gcggtgcttt cggcgtcggc acggacgtcg agctgatcag cgccgtcccg 8520
acctcggagt cgttcctcga gaggaacttc acccaggacg agatcgccta ctgcaaggcc 8580
gcacccgact tccgcgctag cctcgccgcg cgctggtccg ccaaggaggc cactttcaag 8640
gctctcaaga ccgagtcgaa gggcgccgcc gccagcatgc aggacatcga ggtcgtctcc 8700
acgtcgcagg gcccgactat caagctccac ggcgaggtcg agaagatcgc ccaggccgcc 8760
ggcatcacgg ccttcgaggt ctcgctctcg cactcggagg acgtcgcttg cgccgtcgtc 8820
atcgcccaga agtag 8835
<210> 6
<211> 12477
<212> DNA
<213> Aplanochytrium kerguelense
<400> 6
atggaacaac ataaaactga gaaactttca gcagccgatg aaaagttgag tgagagtaca 60
attcattggc ttgctgcaac agatgtccct tcaaactttg tactcttttc cggacaaggg 120
tatcaatact ttgatgagct ccgggaactg tatgaaactg gcacagatga ggttaaaggc 180
ttccttttac ttgcttcgaa aacactgcag gaagagatct gtagtgaaaa agcgaaaact 240
gtcgctttca aatttgtcaa tgagcttgac tttacgaaat ggatagaaga ggatatcgat 300
gtggatcaaa cctacatctt ttctgctcca gtttctttcc ccttaatatt tgtcgctcaa 360
gttgcaaggt acttgcaaac tttaaacttg ctaagaacag accacaaaca atttatccag 420
acaattaaag ggtctagtgg tcatagtcaa gggattgttg ctgctgtatt gatatctaca 480
tcgcctgaca acgacagaat tgttgagaat gcagtcaaac tttgcagata tatgttatgg 540
caagggctga gatgccatga aagcagtgta aacagattca gtggcaaaaa ccacaaatcg 600
aaaaagtcta aagtttattc ctttgatgaa tcgacccgca tggtaaagta ttctcccatg 660
cttgctgtca atggtctcac ggaatccctc ctcaacgatt ttatgaagaa aactgaatcc 720
attattgaag caagaagaac gatacaacaa aagtataacc gaagagaatc attttttgaa 780
acaccaacag aaaaggatga gaaaagtaaa agacttaata tgtttcaaat tgcacttgcg 840
aatggcccca agtcatttgt tgtctctgga gcgccaaaaa cacttattga gctagaacaa 900
agaataaagg agtcaactac atcgtctcca aactctcaaa cacgtatacc atttagcaag 960
cgcaagccag ttgttaaaat gtatttcctc aaggttgggg ctgcttttca tactgaaatt 1020
tgtaatgacg cctctagtaa actaaaagcc gatgtggtcc aaatgaaatt gagtatctcg 1080
ccaaaggaaa tattaattcc tgtctaccat acaaaagatg gttcgaattt atcggagatc 1140
tcagcagatg tgaatgttgt cgatttgcta attgatatgc aaactagtca agtcaatgat 1200
ttcagaaaaa ccttgaagag tatttcaagt cagaatgggg tgtcaaacgt tattgatttt 1260
ggtccaggtg acggaactgc taaattgtgc ctgaaacgaa agtctggaac tggtatacta 1320
gttgtcgcgg ctactggacc actacgaaac agaagatgtg catatggtag aaatcttgtc 1380
ggtctaaact ctgttttgct ggaaaaaaat cctatactag ggatgaattg gggggaagaa 1440
tttaagccca gactgtcgag tcgcaagaat gacaaccaaa tcattgtcgg aactagattt 1500
agtgacttga ctggaaagcc gcctgtaatt ctacctggaa tgacgcctac tacatccttt 1560
cacggcatcg atttggttgc cgcctgtagt aatggtgggt atcatggcga acttgctggt 1620
ggtggattgc cactgccaga atacttcaaa gcgaaaattg atgagctagt gagcaagcaa 1680
aactctggag taggcataaa cataaacatg ctttacctaa actcctactt atggggtttt 1740
caattttcca ctgccattca gatggcgaaa gatggatatc ccatagaatc tatcacttgc 1800
gccgccggag ttccaactac tgaaaaagcg aaagaaataa tgcatcagtt gaaagaagct 1860
ggaattaaat acattgcctt caaacctgga tcaagagccg caattatgga tgtattatct 1920
attgctagag aaaaccccaa tagttcaatc gtactgcaat ggacaggagg acgtggaggt 1980
ggacatcact ccttcgaaga ctttcatgct cctttgctcg acacttatca agaaattaga 2040
gaacacccaa acgttgttct cgtcgtaggc tccggatttg gagatgcaga gaagtcatac 2100
ccatatttag atggttcgtg gtctacattg cctccacata actgtgaatc gagaatgcca 2160
tgtgatgcag ttttatttgg atctaggtgc atggttgcaa aagaagctgc tacctctcca 2220
gaagttaagc aattgattgt taacgctaag ggcgtccgag aaaacgaatc ttgggaaatg 2280
tcttacgaaa acgatgctgg tggaattctc actgtaactt ccgaactcgg cgagcctatc 2340
cataagataa acaaccgcgg gatgaagcta tggagagagt ttgacagaaa gtacttctca 2400
ttaccttcgg gccaagaaag ggaaaaggct atagtgaaag ataaaaagga gataattaga 2460
aggattaatg cagattttca aaaagtttac tttggaagga aggccgatgg aactgttgtg 2520
gacattgcat atatgacata tggtgaaatt ttacggagaa tggtggacct aatgtacgta 2580
accggaggag gagatggaaa acaacataga tttgcgccaa acagatggat agacgttacg 2640
taccaaacta gaacgttcaa gtttttagtt cgcacagaaa gacggttttt gagagacaaa 2700
aaaatggcct ttgcaactga tatgagaaaa ctcgaatctt ttccgataaa gtgcattgat 2760
gagttcgtca agtgctaccc tcaagtagat aaagtgctgg tatcggacga agatgctgga 2820
tattttatcg aactttgtcg aaaccttaga aacggaaaac cggtaaattt tattcctcaa 2880
attgatggct ctttagctta ctggttcaaa aaagactctc tctggtgttc tgaacagatt 2940
gacgcggttc ccgacaatga ccctggcaga gtttgcatac tacatggccc agtagcagcg 3000
caatattcag tagtgtcaga tgagcccata tgcgaaatcc tcggtaacat tcacaaggga 3060
tacgtggaca agttgaaaga ggagaagtat gaggtggcca aagttgaaac aatcggaaag 3120
ctacctgaat acaaaggttc caacaagtct ttccgcaaat ttcttcgtta tggttggttc 3180
aatgccctgt ttcaaagtac cgtaatggtc aaggagaaaa agtgggtcag aaacattatt 3240
ccaggaatca tacaaacgga aaacgaaaac attgtattct cagagcgaac agcaatgaat 3300
gatatgtgtc tcatcgaagt tcttttaaat gattcacaga gaactctagc tttcatagag 3360
tttgaaggaa agcacatttc tgtgtctttg caggattctg aacctgtaaa cagcacactg 3420
accctgtatt ttgagtacca tcctgaaact cctcactctc cgttatttca agtcacaaat 3480
ggaagcaaaa gtattgctag atcaatcaaa cagtattaca ggaatatatg ggattgtcag 3540
gatgttgttt ctatccatga tacattttcc gaaagcttca ccgtcacgag agaagacatc 3600
gaaaggttca atggcgcaat tcaatatgcg gtaaatactt cggagggaac tttagacttt 3660
gcaattgttg caggatggaa atcactgata aaagctctgt tttcaaaaga gatagacggc 3720
agtctgttag atcttgttca catctcccac tcatacaaac tacttgtaga ggagaaggag 3780
agaacgcttg ttcaagctgg agagaaaatt tgttcggaat ctcgcgttat ttcagtacag 3840
atattaccag gtgttggaaa aagtataaca gttaaagggg tactctctag aaacacagtc 3900
aaatgggttg aagtccaatc cgagttttta atccgaggca atttcttcga ttttcaaaag 3960
actttcaagg aatatagcta caaagctgag gttttatgta aagacgatac agttccgcaa 4020
atcctaaact cacaggaatg gctttttctg gactctggag tcagacctct atctaaacac 4080
gataaagttg tatttcaaat cgatcgcgtt gtggaaaaga gaaagtctgt gacaaatatt 4140
tctgatattt tagttgctgg taatgttgtt cgaatcaata cacctgaaga gatcgatggt 4200
tttattgaat tgaaagacca aggaagccca aagggaatca tcattgggaa agttgattta 4260
agccttgaaa atgccaatct gagtgaaaat ccaattatta catatttgga ttccattaaa 4320
gaatcaaaat tgcacggtag tgtttttgaa tcgggaggat atactgtgat gccgaggccc 4380
gaactagtaa cagcgccaag tattgtcgaa agcattgtgt atgcatctgc ttctagagat 4440
atgaatccaa ttcatcggaa tcgcacattc gctaagttag ctggactccc cggtggtagt 4500
accattgtgc acggaatgtg gacagctgca atgagcagaa gaattctaga attacataca 4560
gttctcggag accatcgccg tatttccaca tactcggtta aatttaccgg catggtgttc 4620
ccagaagaca aactagccgt tatggtcaaa cacgttggtg ttacggaagg tagacttatt 4680
cttgatgtag atgttagcaa agtagaaacg caagagcggg ttctccacgg acgagctgaa 4740
gtagaagggc cgtcgacatc attcctattc actggtcaag gatccgcaaa agttgggatg 4800
ggaatggaca gctacgaaga aaagcagtcg gcacgggcag tttggcaaac tgcagataaa 4860
tttttgagga aaaaatttgg attttcaatt gttgatattg tcaaaaataa tccaaaagag 4920
ttgactgttc gtttcggtgg atcacaaggg aaagtgattc gagaaaacta cagagccatt 4980
aaaagggaag atggatctca attgatccga gaaatccata gtaataccac gtcatttaca 5040
tttcgttcac caaatgggct tttatttgcc acgcaatttt ctcagccggc gctggtacta 5100
gtgcaaaaag cagcgtttga agaaatgctt caaggagggt ttgttccgtc aaacagtatg 5160
tttgctggtc attctcttgg tgaatacgca gctcttgcct cgtatgcaaa cgttctgagt 5220
atcgaagact tagttgaaac tgtattttta agaggaatgg taatgcaaaa tgcagttgca 5280
agggacgacg aaggaacatc tgattacggt atggtggctg ccaatccttc aagagtcagt 5340
aaaacgttct cggcacaatt gttgatggaa atcgtccaga aaattgacga cgaatcagat 5400
gacagtcatc ttctccaagt agtaaatttc aatattagag atgctcagta cgtaatagct 5460
ggaaatctgg aattgttaga tagtctatcg aatgttctca atgtaatttc aaacagccac 5520
gacaaaacct ttgacagaag cattatccaa gctgcaatga taaaggctga aaataggaga 5580
agaagctgca aggaaaacaa tcaacctttc cgtcttaagc gtggaatcgc aaccattcct 5640
cttcacggca ttgatgtacc ttttcattct cgacagcttt tgaatggggt acctgctttt 5700
cggtctctac tagaaccaag atttacggaa gaaatgattc aaaagtactt gcctaggttg 5760
atcggaaagt acattcctaa tgtgcacgca gaaccgtttt ccattagcaa agagtacatt 5820
gaaaaggttg caagtgtaac aaaatcgcca gcgttgtaca agcttctatc tacatactca 5880
acaatgtgtg actcatcaaa agcaagaatc ttgctcatag agcttttagc tcatcaattt 5940
gcaatgccag ttcaatggat agatacccag gactatatac tctccaggca cacacagcga 6000
ataattgaga tgggcccagc cccaactctt gttggtatgg cgaaaaagac attgcaatct 6060
agaatatatg gtgacgaaga agactacaaa cctagtattc tgtggtggaa tcaaaaccag 6120
gaggaaatct tctataactt ggacaatcaa ggaatatcta ttcaacagtt tttgcaagaa 6180
ttaaaaggaa ctgaagatga cgagtcacag tctgatgata tgtcatcata tacccagtca 6240
gacgagacta tacaaagaac tgcaaagaac agggttatgc cagaagcaaa tgcaaacgac 6300
aaggagaaag tggggaatga acttcaaaag aaggaagtta caaacaggcc tcttcatgtc 6360
ctaagagtcc tcatctcaac caaacttaag aagaaaatgt tggaggtaaa ggcaacagac 6420
accatcaagg gtgtttctgg tggaaaaagt gcgattcaaa atgagctaat tggagaaata 6480
aacgctgaat acagcagttt tagagatatt gaagatattt cggaagtttc tttagctgag 6540
ctagctagaa gggccccatc ccggcaggaa acagttcttg gtaaagttac aacgaaaatg 6600
gtgaacaaaa tgttttctag caaactgccc ggaaaatttg gtcctaatga tactaaggag 6660
tacttacaaa gcaaactctc ggatgcgtcc acagtcgatt cagtgtctct tcatgccctg 6720
actctgcagc cggtttctag actagctacc cctggagagg cacaggcatg gcttgattct 6780
gtttgtagag actttgaaaa cgatcagaga atcagtattc taaagaacaa aagtccaaat 6840
tcgaacgaaa gtgtgatgat ttctgaacaa aggactgttc aaaacaactt tgtacaaatt 6900
aagactatat tgatattggg tgactcattg tccgctggtt atggtattaa tcctgaacaa 6960
ggttgggtcg ccttattgca aaagagattg gatcaacaat tcccaaagca acataaagta 7020
atcaatgcat ctgtttcagg tgaaactaca tctggtgctt tggcaagatt accaaagttg 7080
ttaaccactt acagacctaa cgttgtcgta attgaattgg gtggtaacga cgccttaaga 7140
ggtcaaccac ctcaaatgat ccaatcaaat ttggaaaagt taatacaaca ctcccaaaaa 7200
gctaagagta aggttgtcgt attcggtatg aagatcccac ctaactatgg tacagcatac 7260
tctcaagcct tcgaaaataa ctataaggtt gtctcacaaa cctaccaagt caaattgtta 7320
ccatttttct tggatggtgt tgctggtcat aagtccttaa tgcaaaatga ccaaatccac 7380
ccaaacgcca aagctcaaag tatattgttg aacaacgctt acccttacat caagggtgca 7440
ttacgtactc tctcttcagg tgatagtagt cctgctcatt cgacaaaaga tatcctttcg 7500
tccactgcaa tgaagaaatt gaaaaaattg tacttggacc aggtggatgt cttcaacact 7560
tttttgggag aggatcgacg ggccaaactt caaaaggttc taacagagga aaccagtgca 7620
aaagatagaa tagaggctca gctccttgaa ttaaaggaag aacttggaga ccctttttgt 7680
gacgggatac aaccccagtt taatgaagct cggatacgaa tctatgattc atactggaac 7740
tgggtggtgc aggatgcatt agaactccat tatcatacat tctcttgtgt gttgaactca 7800
aaaggcaaga acgtaaatat tcccaattca tctaacagct attttcgcgc aatgagcgac 7860
tggataactt cttcaaacga aaagttggaa gaaaataaac ctccgcaggc ttggtttcgc 7920
aattaccttt gtaatcgtgc cactccagaa cttctaaccg ttgtacaata ctttgcagca 7980
agaatgaacg aacaaggaca ttcagaatat gctcaagcag tatctctttt agcagaacaa 8040
gtatcacagt ggatatctag accacctgta cacatagcat tgtttgcatc tctagaacct 8100
agggtgacag ttgatagtgc taacaatttt ggattgaaat atgaagaaaa accaagaaag 8160
tttgtggctg tcggtgaaaa caaagctttc tcgtgtgata atgcgtccct gtatgttaaa 8220
gaaatgagca aggggctatt ttatgaccac cgtgtggctt cgaaagttga acatcccagc 8280
caaagtgtat acctttctcc cgataattct ttctttccaa acgaggaaga gagtgaccag 8340
atttcaagcg gaatgagact accaaaaaca caaggagaaa tccagcgtga tttttcaaga 8400
ctccctgctg gtcagaagct agaagtcatg agaaagagtg ttaatcggtc gtccgacaca 8460
gatgaggaaa ttgccaacat cgttcgacaa aactataaca gtatccatgt ggcaaaaaac 8520
gtacctttgg tgcatttgaa atcgccttct aaatttgaca agacggtaag agtactcgat 8580
gagcctttga catcaatgta cttgagttgt ttgcatgata ttgcaacaag tggagttagc 8640
tttgcaggac aaaacgcttt ggtgacagga gccggctttg gttctattgg gattgaatta 8700
atcaaaccgt tgcttgaggg gggggcaacg gttttagtaa cagttagact taatcggact 8760
gacgagcaaa tgcaaattgt taatgaaagg tttcaaagat tgtacgaaga gtttgggtcc 8820
agaggaagca agctcgttct ggttccttgc aattgcgctt caaatcaaga tgtacattcc 8880
ctcattagac acatctatga gaaattgaaa cttgacctag attttatatt tccatttgct 8940
gctatcgggg agcaagggaa agatgtgtct gaaattggct ccaaatctga agtcgctcat 9000
agactaatgc tgactaatac tataagatta ctaggtgcag tgaaaaaggc caaagaggat 9060
agatgcattg aaacaaggcc tgcactggtt ctgttgccat gctctccaaa ccatggggat 9120
tttggactag atgggttata tgcagaatca aagcttggat tggaatccct agtaaacaag 9180
tggaaaagtg aagaatgggg aaattaccta agtatttgcg ctgcagtcat aggttggaca 9240
aggtccaaac tcatgtggca gaacaatgtc gttgccgagg gaatagaaaa gttgggagtg 9300
agaacctttt ctactacaga gactgctttt aacctgattg gtctactaca tcctgatatc 9360
gtgtctcatg cagcagaaga acctctatgg gcagacctta ctggaaactg gggcgctgct 9420
cctgacttga aggaacattc aaaaaggatt cgaacgtcac ttctggcaca aagtaaagca 9480
gcaaaagcca tatcgctgtc gtcaaaatcg ctcgttatag aaccttctga atccaagacg 9540
aaagcgcaga cagaaattgt taacacatac ctagctggtg tgaatgataa gttgaggctt 9600
ccgctagcaa atcccgaaaa gttctgcaat ccattcccta aaattccatc ccaggaacga 9660
atggattcat tagcatacct taaacactca gtagatctca aaaaagtcgt agtagtagta 9720
ggatatggag aaattgggcc ttggggaaat tcgcgcacta gatgggagat ggagtcattc 9780
ggtgaattct cgcttgaagg tgcaatcgaa cttgcttggc tcgttggctt aatcaaacca 9840
gttactggac cgttaaaaaa tgatcctaga acgcaatact ttggttgggt agatgcagaa 9900
tcagaagagc cagttgcaga tcatgaaata aagactcgat atgagaaagt actgcttcag 9960
cactctggga ttcgtcttat tgaaccggag ctctttgaag gatataatcc aaagaaaaag 10020
tcaattctga gacaagtagc catcgctgaa gacatgaaac caatagaagt agcttctttg 10080
gaagaagccc agcagtacgt gaatgagtta gggaaagagt tcatagacgt ttttaatgaa 10140
aacgttgaat ctaatgatgg acaatggtat attcgcctaa aagcaggagc agtggtctcc 10200
attcctggag cgttaagctt caacagattt attgctggtc agctacctac aggatgggat 10260
gcaaaacgac taggaatacc agatgatatt gccgactctg tagatcctgt tacgctatat 10320
gctctggttt caacagttga agccttggtg tgtgctggat tgacagatcc atatgaactg 10380
tatcaatatg tgcatgtaag tcaagttgga aacacgtcag gtggcggcat gggagggatg 10440
cgatcattga agcgaatgtt cctagaaaga aaacttgatg cagaaatccc ttctgatact 10500
ttagcagagt catttattaa cacaatgcca gcctgggtga acatgctgct tctgtcaagc 10560
tcgggaccaa taaagactcc agtcggtgct tgcgccacag cggcagagtc ggtagacata 10620
ggaattgaaa ccatcttagc aggaaaagcc agagtcgtta tagcgggcgg gtatgatgac 10680
ttttgtgaaa cgggaagtaa tgagtttgcc atgatgggag caacttccaa cagtcaaaca 10740
gaagctgaaa aaggcagatt tccacgtgag gcatcaagac caatgacaga tacacgtgcc 10800
ggattcatgg aatcacaagg cgctggtatg caagtgttaa tggatgctga attagccatt 10860
cagatggggc ttccagtata tggtattctt gctctgtcaa acactgctac cgatagacaa 10920
ggaagatccg tgcccgcccc aggtcgtgga atactaacca ctgcgagaga agtgcgctca 10980
aaaaactcta aaagtagcca caacggcaaa ccagtagaaa atcctctatt gtctgtaaag 11040
ttccgcaaaa aacatttgag acaagagctc gatgctatag atctgtgggc cagccaagag 11100
atatcgaaca tgtctgagga attctcacag aattcaagaa aaatggaaaa caggaaaggt 11160
tttgtagaaa caatgcgaaa aaagaagcgt tcggcagctt ttgaaacctg gggacaagga 11220
ttctatagga atgatgactc catagcccct ttacgcggag ctttgtccgt atggggatta 11280
acagttgatg atcttcttgt gggaagtttt cacggaactg gaaccaatct taacgatacg 11340
aatgagtcct ccttagtgaa taagcagctt aagcatttgg atcgaaaaga aggaaacatt 11400
ctgttagtag taactcaaaa atatctgact ggtcatccaa agggtgcagc tgctgcctgg 11460
atgctcaatg gattgctgca atgcatgaat tctggaagag taccgggaaa ccgaaacttg 11520
gacaatgtag atggaaagct cagaacgaac ggttacttgt tctatccaaa tcggacaatt 11580
gaggttccaa gagtggaagc agcttttctt aaaagctttg gatttggtca agccggagct 11640
gaagttgtca ttattcatcc tgatcgtctc ctggctgttt taagtgaaga aaatctgaaa 11700
tcgtatatac ttcgaagaaa tgaaagagag aaaagagcat atcgatatca tcaaggcgta 11760
atgagtggac atcatactat ggtgcaagtg aaggaatttg ctccatatga agacgatatc 11820
ctagaagaaa tatatttgaa tccaagagct agggcttcct ttgactcatc gaaaagtact 11880
tggacgtttc acaaatatag gaacgacaaa gatactgtag acgataaaga tgaaatggag 11940
caagattcat tgaatgaatc agtagatctt ccgaaagata ttacaaaagt tggatcaccc 12000
tctgaaatac gtattccagt aaaaacaaga ttggaagtta ctgttcgtga aggagtcgaa 12060
ggtttgacga aaaaggacaa gttttcaagt caaggggttg gcgttgatgt tgaaccagtt 12120
tcaacttttg cacaacatga agagaaaacg atctttattc agaacaactt cactgagaat 12180
gaacagctgt attgtaatca tgctgcaagt ccagcagcaa gctatgcggg tagatgggca 12240
gcgaaggaag cagtgattaa agccatcagc aattcttcac ttgaaacaag atcactttgg 12300
caaggagctg agggtaaact gatcgacatt gaaatcattc aaagtaattc tggtgctcct 12360
gaggtagtct tgcatggtca tgcaaaagaa gtattccaaa ctttaggtct tacaaatgtt 12420
aaggtttcta tcagtcatac tccagaagtt gcagtagccc aagcaataac aaattaa 12477
<210> 7
<211> 543
<212> DNA
<213> 白锈菌
<400> 7
aagactatat tgatattggg tgactcattg tccgctggtt atggtattaa tcctgaacaa 60
ggttgggtcg ccttattgca aaagagattg gatcaacaat tcccaaagca acataaagta 120
atcaatgcat ctgtttcagg tgaaactaca tctggtgctt tggcaagatt accaaagttg 180
ttaaccactt acagacctaa cgttgtcgta attgaattgg gtggtaacga cgccttaaga 240
ggtcaaccac ctcaaatgat ccaatcaaat ttggaaaagt taatacaaca ctcccaaaaa 300
gctaagagta aggttgtcgt attcggtatg aagatcccac ctaactatgg tacagcatac 360
tctcaagcct tcgaaaataa ctataaggtt gtctcacaaa cctaccaagt caaattgtta 420
ccatttttct tggatggtgt tgctggtcat aagtccttaa tgcaaaatga ccaaatccac 480
ccaaacgcca aagctcaaag tatattgttg aacaacgctt acccttacat caagggtgca 540
tta 543
<210> 8
<211> 1266
<212> PRT
<213> 圆红冬孢酵母
<400> 8
Met Asn Gly Arg Ala Thr Arg Ser Val Thr Gly Thr Ser Thr Pro Val
1 5 10 15
His Thr Ala Thr Thr Arg Pro Leu Val Leu Leu His Pro Ser Thr Gln
20 25 30
Thr Arg Ile Ser Leu His Val Pro Ser Thr Ser Gln Glu Trp Ile Ala
35 40 45
Ala Glu Val Ala Arg Asp Thr Phe Gln Asp Trp Leu His Ala Ala Glu
50 55 60
Lys Ser Gly Asn Leu Val Gly Phe Glu Ala Ala Glu Leu Asp Asp Glu
65 70 75 80
Gln Ala Gly Glu Gly Asp Asp Glu Lys Glu Leu Val Leu Thr Ala Tyr
85 90 95
Phe Leu Lys His Val Ala Gly Leu Leu Pro Phe Pro Ser Thr Ala Thr
100 105 110
Ser Pro Ala Thr Ala Ala Val Leu Leu Ala Ala Phe Asn His Phe Ala
115 120 125
Ser Val Tyr Leu Ser Gly Thr Asp Val His Thr Leu Thr Ala Ser Leu
130 135 140
Ala Ala Pro Val Arg Ala Leu Val Ile Ser Ser Phe Phe Leu Ala Lys
145 150 155 160
Thr Lys Leu Glu Val Glu Gly Leu Gly Lys Val Leu Pro Lys Gln Ser
165 170 175
Glu Ser Ala Leu Leu Gln Lys Ala Ala Thr Gly Gln Ala Glu Val Phe
180 185 190
Ala Leu Phe Gly Gly Gln Gly Met Asn Glu Val Tyr Phe Asp Glu Leu
195 200 205
Gln Thr Leu His Asp Leu Tyr Thr Pro Leu Leu Thr Pro Phe Leu Ala
210 215 220
Arg Ala Ser Glu His Leu Val Ser Leu Ala Ala Ala Glu Gln His Thr
225 230 235 240
Leu Leu Tyr Asp His Ser Leu Asp Ala Leu Ala Trp Leu Gln Asp Pro
245 250 255
Ser Thr Arg Pro Glu Val Pro Tyr Leu Ala Thr Cys Ala Val Ser Leu
260 265 270
Pro Leu Ile Gly Leu Thr Gln Leu Cys Gln Tyr Val Val Tyr Gly Lys
275 280 285
Gly Ser Ser Leu Gly Pro Ala Glu Leu Gly Ala Lys Phe Lys Gly Ala
290 295 300
Thr Gly His Ser Gln Gly Val Val Ser Ala Leu Val Ile Ala His Glu
305 310 315 320
Tyr Pro Pro Ala Ser Lys Asp Gly Ser Asp Ala Trp Glu Pro Phe Tyr
325 330 335
Glu Gln Ala Leu Arg Gly Leu Thr Val Leu Phe Gln Ile Gly Leu Gln
340 345 350
Gly Thr Leu Ala Phe Pro Ser Ile Ala Ile Ser Pro Ala Leu Glu Ser
355 360 365
Ser Ser Val Glu Asn Gly Glu Gly Val Pro Thr Ala Met Leu Ala Val
370 375 380
Thr Gly Leu Asp Leu Lys Ser Leu Glu Lys Lys Ile Ala Glu Val Asn
385 390 395 400
Gly His Val Lys Ser Glu Gly Arg Asp Glu Thr Val Ser Ile Ser Leu
405 410 415
Tyr Asn Gly Ala Arg Ala Phe Val Val Thr Gly Ala Pro Lys Asp Leu
420 425 430
Val Gly Leu Ala Asp Gly Leu Arg Lys Asn Arg Ala Pro Ala Gly Lys
435 440 445
Asp Gln Ser Lys Ile Pro His Ser Lys Arg Leu Pro Val Phe Ser Met
450 455 460
Arg Phe Leu Pro Ile Asn Val Pro Tyr His Ser His Leu Leu Gln Gly
465 470 475 480
Ala Thr Glu Lys Ala Leu Ala Thr Phe Ser Ala Glu Glu Ala Ala His
485 490 495
Trp Ala Pro Ser Ser Phe Thr Cys Ala Val Tyr Asn Thr Glu Asp Gly
500 505 510
Ser Asp Met Arg Gln Leu Ser Ala Ser Ser Val Leu Glu Ser Val Phe
515 520 525
Gln Gln Ile Phe Thr Ser Pro Ile His Trp Val Ser His Ala Thr Asn
530 535 540
Phe Pro Ser Ser Ala Thr His Ala Ile Asp Phe Gly Thr Gly Gly Ala
545 550 555 560
Ser Gly Ile Gly Ser Leu Cys Ala Arg Asn Trp Glu Gly Arg Gly Ile
565 570 575
Arg Thr Ile Met Leu Gly Asn Arg Gly Glu Gly Val Gly Ala Gly Lys
580 585 590
Glu Ala Trp Gly Lys Lys Val Pro Thr Glu Glu Lys Trp Asn Glu Arg
595 600 605
Phe His Pro Arg Leu Val Arg Thr Ser Asp Gly Lys Ile His Leu Asp
610 615 620
Thr Pro Phe Ser Arg Leu Leu Ser Lys Pro Pro Leu Met Val Gly Gly
625 630 635 640
Met Thr Pro Thr Thr Val Lys Ala Gly Phe Val Ser Ala Val Leu Arg
645 650 655
Ala Gly Tyr His Ile Glu Leu Ala Gly Gly Gly His Tyr Asn Glu Lys
660 665 670
Ala Val Arg Ala Lys Val Ala Glu Ile Gln Lys Leu Val Asn Lys Pro
675 680 685
Gly Met Gly Ile Thr Leu Asn Ser Leu Tyr Ile Asn Gln Arg Gln Trp
690 695 700
Thr Phe Gln Phe Pro Leu Trp Ala Lys Met Lys Gln Glu Gly Glu Pro
705 710 715 720
Val Glu Gly Leu Cys Val Ala Ala Gly Ile Pro Ser Thr Glu Lys Ala
725 730 735
Lys Glu Ile Ile Asp Thr Leu Arg Glu Ala Gly Ile Lys His Val Ser
740 745 750
Phe Lys Pro Gly Ser Val Asp Gly Ile Arg Gln Val Val Asn Ile Ala
755 760 765
Ser Ala Asn Pro Asp Phe Pro Ile Ile Leu Gln Trp Thr Gly Gly Arg
770 775 780
Ala Gly Gly His His Ser Cys Glu Asp Phe His Ala Pro Ile Leu Ala
785 790 795 800
Thr Tyr Ala Ser Ile Arg Gln His Pro Asn Ile Lys Leu Val Ala Gly
805 810 815
Ser Gly Phe Gly Ser Ala Glu Gly Cys Tyr Pro Tyr Leu Ser Gly Glu
820 825 830
Trp Ser Glu Lys Gln Tyr Gly Val Ala Arg Met Pro Phe Asp Gly Phe
835 840 845
Met Phe Ala Ser Trp Val Met Val Ala Lys Glu Ala His Thr Ser Glu
850 855 860
Ser Val Lys Gln Leu Ile Val Asp Ala Pro Gly Val Glu Asp Gly Gln
865 870 875 880
Trp Glu Gln Thr Tyr Asp Lys Pro Thr Gly Gly Ile Leu Thr Val Asn
885 890 895
Ser Glu Leu Gly Glu Pro Ile His Lys Val Ala Thr Arg Gly Val Lys
900 905 910
Leu Trp Ala Glu Phe Asp Lys Lys Val Phe Ser Leu Ser Lys Glu Lys
915 920 925
Gln Leu Ala Trp Leu Ala Asp Asn Lys Lys Tyr Val Ile Asp Arg Leu
930 935 940
Asn Ala Asp Phe Gln Lys Pro Trp Phe Pro Ala Lys Ala Asp Gly Ser
945 950 955 960
Pro Cys Asp Leu Ala Asp Met Thr Tyr Ala Glu Val Asn Ala Arg Leu
965 970 975
Val Arg Leu Met Tyr Val Ala His Glu Lys Arg Trp Ile Asp Pro Ser
980 985 990
Leu Arg Asn Leu Val Gly Asp Trp Ile Arg Arg Val Glu Glu Arg Leu
995 1000 1005
Ser Asn Val Asn Asp Ser Gly Ile Lys Ile Ser Ala Leu Gln Ser
1010 1015 1020
Tyr Ser Glu Leu Asn Glu Pro Glu Ala Phe Leu Lys Gln Phe Leu
1025 1030 1035
Ala Gln Tyr Pro Gln Ala Glu Asp Gln Ile Leu Ala Ser Ala Asp
1040 1045 1050
Val Ser Tyr Phe Leu Ala Ile Ser Gln Arg Pro Gly Gln Lys Pro
1055 1060 1065
Val Pro Phe Ile Pro Val Leu Asp Ala Asn Phe Ser Ile Trp Phe
1070 1075 1080
Lys Lys Asp Ser Leu Trp Gln Ala Glu Asp Ile Glu Ala Val Phe
1085 1090 1095
Asp Gln Asp Pro Gln Arg Val Cys Ile Leu Gln Gly Pro Val Ala
1100 1105 1110
Ala Lys His Cys Thr Ser Thr Gln Thr Pro Ile Ala Glu Met Leu
1115 1120 1125
Gly Asn Ile Glu His Gln Leu Val Lys Asn Val Leu Asp Asp Tyr
1130 1135 1140
Tyr Gly Gly Asp Glu Ser Gln Ile Pro Thr Ile Asp Tyr Leu Ala
1145 1150 1155
Pro Pro Pro Lys Pro Val Asp Ala Gly Ala Ile Leu Ala Glu Asn
1160 1165 1170
Asn Ile Ala His Ser Val Glu Glu Leu Ala Asp Gly Gly Lys Lys
1175 1180 1185
His Val Tyr Ser Ile Asn Gly Val Leu Pro Pro Thr Gly Asp Trp
1190 1195 1200
His Ala Ala Leu Ala Gly Pro Lys Leu Asp Trp Leu Gln Ala Phe
1205 1210 1215
Leu Ser Asn Val Ser Ile Gln Ala Gly Glu Gln Ser Ile Pro Asn
1220 1225 1230
Pro Val Lys Lys Val Leu Ala Pro Arg His Gly Gln Arg Val Glu
1235 1240 1245
Leu Thr Leu Asn Lys Asp Gly Gln Pro Leu Lys Leu Asp Val Phe
1250 1255 1260
Gly Gly Leu
1265
<210> 9
<211> 2928
<212> PRT
<213> 圆红冬孢酵母
<400> 9
Met Val Ala Ala Gln Asp Leu Pro Leu Ala Leu Ser Ile Ser Phe Ala
1 5 10 15
Pro Glu Ser Ser Thr Ile Ser Met Thr Leu Phe Asn Gln Pro Glu Ala
20 25 30
Ser Lys Pro Ala Leu Pro Leu Glu Leu Lys Tyr Lys Tyr Asp Pro Ser
35 40 45
Thr Pro Tyr Ala Pro Ile His Glu Ile Thr Glu Asp Arg Asn Gln Arg
50 55 60
Ile Lys Gln His Tyr Trp Asp Leu Trp Gly Leu Gly Asn Lys Ala Asp
65 70 75 80
Gln Gly Ile Ser Gln Leu Lys Ile Thr Asp Glu Phe Gln Gly Asp Leu
85 90 95
Val Thr Ile Ser Ala Asp Glu Ile Glu Ala Phe Cys Arg Val Val Gly
100 105 110
Ile Glu Gly Glu Ala Tyr Lys Arg Asn His Lys Ala Gly Met Gln Val
115 120 125
Pro Leu Asp Phe Ala Ile Lys Leu Gly Trp Lys Ala Ile Met Lys Pro
130 135 140
Ile Phe Pro Ser Thr Ile Asp Gly Asp Leu Leu Lys Leu Val His Leu
145 150 155 160
Ser Asn Gly Phe Arg Val Leu Pro Asp Thr Pro Thr Leu Gln Val Gly
165 170 175
Asp Val Val Thr Thr Thr Ser Arg Ile Glu Ser Ile Thr Asn Ser Asp
180 185 190
Thr Gly Lys Thr Val Ser Val Arg Gly Val Ile Ser Leu Val Ser Ser
195 200 205
Ala Asp Ser Lys Gly Lys Asp Ala Ser Thr Glu Asp Arg Ile Pro Leu
210 215 220
Ile Glu Val Thr Ser Ser Phe Phe Tyr Arg Gly Lys Phe Ser Asp Tyr
225 230 235 240
Ala Gln Thr Phe Ser Arg Val Ala His Pro Thr Tyr Ser Val Pro Ile
245 250 255
Thr Thr Pro Glu Ala Val Ala Val Leu Gln Ser Lys Glu Trp Phe Gln
260 265 270
Trp Asp Asp Asp Ser Lys Pro Leu Glu Val Gly Thr Lys Leu Gln Phe
275 280 285
Lys Val Glu Ser Asn Tyr Val Tyr Ala Asp Lys Ser Ser Tyr Ala Met
290 295 300
Ala Thr Val Thr Gly Gly Ala Tyr Val Ile Thr Pro Glu Leu Lys Leu
305 310 315 320
Ala Val Lys Val Ala Thr Val Asp Tyr Thr Ser Glu Gly Glu Gly Val
325 330 335
Ile Gln Gly Asp Pro Val Ile Glu Tyr Leu Lys Arg His Gly Ser Ala
340 345 350
Leu Asp Gln Pro Ile Met Leu Glu Asn Gly Gly Tyr Ser Leu Thr Lys
355 360 365
Ala Gly Gln Cys Thr Phe Thr Thr Pro Ala Ser Asn Leu Asp Tyr Ser
370 375 380
Leu Thr Ser Gly Asp Thr Asn Pro Ile His Thr Asn Pro Tyr Phe Ala
385 390 395 400
Ser Leu Ala Tyr Leu Pro Gly Thr Ile Thr His Gly Met His Ser Ser
405 410 415
Ala Arg Thr Arg Lys Phe Val Glu Gln Val Ala Ala Asp Asn Val Gly
420 425 430
Ala Arg Val Arg Lys Tyr Glu Val Gly Phe Thr Ala Met Cys Leu Pro
435 440 445
Ser Arg Lys Met Glu Val Arg Leu Lys His Val Gly Met Thr Ala Asp
450 455 460
Gly Asn Arg Leu Ile Lys Val Glu Thr Val Asp Val Glu Gly Gly Asn
465 470 475 480
Val Val Leu Ser Gly Thr Ala Glu Val Ala Gln Ala Pro Thr Ala Tyr
485 490 495
Val Phe Thr Gly Gln Gly Ser Gln Glu Pro Gly Met Gly Met Glu Leu
500 505 510
Tyr Ala Asn Ser Pro Val Ala Arg Ala Val Trp Asp Glu Ala Asp Arg
515 520 525
His Leu Gly Glu Val Tyr Gly Phe Ser Ile Leu Glu Ile Val Arg Thr
530 535 540
Asn Pro Lys Glu Lys Thr Val His Phe Gly Gly Leu Lys Gly Gln Ala
545 550 555 560
Thr Arg Gln Lys Tyr Met Asp Met Ser Tyr Thr Thr Thr Asp His Glu
565 570 575
Gly Asn Val Lys Thr Leu Pro Leu Phe Gly Asp Ile Asp Leu Arg Thr
580 585 590
Ser Arg Tyr Thr Phe Ser Ser Pro Thr Gly Leu Leu Tyr Ala Thr Gln
595 600 605
Phe Ala Gln Ile Ala Leu Val Val Thr Glu Lys Ala Ala Phe Glu Asp
610 615 620
Met Arg Ala Lys Gly Leu Val Gln Lys Asp Cys Val Phe Ala Gly His
625 630 635 640
Ser Leu Gly Glu Tyr Ser Ala Leu Ala Ser Ile Ala Asp Ile Leu Pro
645 650 655
Ile Ser Ala Leu Val Asp Val Val Phe Tyr Arg Gly Ile Thr Met Gln
660 665 670
Arg Ala Val Glu Arg Asp His Leu Asn Arg Ser Ser Tyr Gly Met Val
675 680 685
Ala Val Asn Pro Ser Arg Ile Gly Lys Ser Phe Gly Asp Ala Ala Leu
690 695 700
Arg Glu Val Val Asp Thr Ile Ala Arg Arg Gly Asn Ile Leu Ile Glu
705 710 715 720
Val Val Asn Tyr Asn Val Glu Gly Gln Gln Tyr Val Val Ala Gly His
725 730 735
Leu Val Ala Leu Gln Ser Leu Thr Asn Val Leu Asn Phe Leu Lys Ile
740 745 750
Gln Lys Ile Asp Leu Ala Lys Leu Thr Glu Thr Met Ser Ile Glu Gln
755 760 765
Val Lys Glu His Leu Cys Glu Ile Val Asp Glu Cys Val Gln Lys Ala
770 775 780
Arg Asp Leu Gln Ala Lys Thr Gly Phe Ile Thr Leu Glu Arg Gly Phe
785 790 795 800
Ala Thr Ile Pro Leu Pro Gly Ile Asp Val Pro Phe His Ser Arg Tyr
805 810 815
Leu Trp Ala Gly Val Met Pro Phe Arg Thr Tyr Leu Ser Lys Lys Val
820 825 830
Asn Pro Ala His Phe Asn Ala Asp Leu Leu Val Gly Arg Tyr Ile Pro
835 840 845
Asn Leu Thr Ala Val His Tyr Glu Val Ser Lys Glu Tyr Ala Glu Arg
850 855 860
Ile His Thr Gln Thr Ser Ser Pro Arg Leu Asn Lys Ile Leu Lys Ala
865 870 875 880
Trp Asp Glu Glu Arg Trp Gly Ala Pro Glu Asn Arg Asn Lys Leu Gly
885 890 895
Tyr Ala Ile Leu Ile Glu Leu Leu Ala Tyr Gln Phe Ala Ser Pro Val
900 905 910
Arg Trp Ile Glu Thr Gln Asp Ile Leu Phe Arg Asp Phe Lys Phe Glu
915 920 925
Arg Leu Val Glu Leu Gly Pro Ser Pro Thr Leu Thr Gly Met Ala Thr
930 935 940
Arg Thr Gln Lys Leu Lys Tyr Asp Ala His Asp Ser Ser Val Gly Ile
945 950 955 960
Lys Arg Ser Ile Tyr Cys Ile Ala Lys His Gln Lys Glu Ile Tyr Tyr
965 970 975
Gln Phe Asp Asp Val Ala Gly Glu Glu Ala Pro Ala Pro Ala Ala Val
980 985 990
Ala Pro Ser Ala Pro Ala Pro Lys Ala Ala Pro Val Ala Ala Ala Pro
995 1000 1005
Pro Pro Pro Ala Pro Val Ala Ala Ala Pro Ala Ala Ala Val Ala
1010 1015 1020
Asp Glu Pro Leu Lys Ala Val Asp Thr Leu Arg Ile Ile Ile Ala
1025 1030 1035
Gln Lys Leu Lys Lys Pro Val Gly Glu Val Pro Leu Thr Lys Ser
1040 1045 1050
Ile Lys Glu Leu Val Gly Gly Lys Ser Thr Leu Gln Asn Glu Ile
1055 1060 1065
Leu Gly Asp Leu Gln Gly Glu Phe Ser Ser Ala Pro Glu Lys Gly
1070 1075 1080
Glu Glu Met Pro Leu Gln Glu Leu Gly Ala Ala Leu Gln Gln Gly
1085 1090 1095
Tyr Ser Gly Lys Leu Gly Lys Tyr Thr Thr Gly Val Ile Ser Arg
1100 1105 1110
Met Ile Gly Ala Lys Met Pro Gly Gly Phe Gly Leu Ser Ala Val
1115 1120 1125
Gln Gly His Leu Gly Lys Thr Tyr Gly Leu Gly Ala Gly Arg Ile
1130 1135 1140
Asp Gly Val Leu Leu Phe Ala Val Thr Gln Glu Pro Ala Lys Arg
1145 1150 1155
Leu Ala Asn Glu Gly Glu Ala Lys Ala Trp Val Asp Ser Val Ala
1160 1165 1170
Gln Gly Tyr Ala Ser Met Ala Gly Ile Ser Leu Ala Ala Gly Gly
1175 1180 1185
Gly Ala Ala Ala Ala Ala Pro Ala Met Ala Phe Ala Ala Pro Ala
1190 1195 1200
Ala Ala Gly Gly Gly Ala Pro Ala Ala Val Pro Asp Glu Pro Leu
1205 1210 1215
Lys Ala Thr Asp Thr Leu Arg Ala Ile Ile Ala Gln Lys Leu Lys
1220 1225 1230
Lys Gln Ile Pro Asp Val Pro Leu Thr Lys Ser Ile Lys Asp Leu
1235 1240 1245
Val Gly Gly Lys Ser Thr Leu Gln Asn Glu Ile Leu Gly Asp Leu
1250 1255 1260
Gln Gly Glu Phe Ser Ser Ala Pro Glu Lys Gly Glu Glu Met Pro
1265 1270 1275
Leu Gln Glu Leu Gly Ala Ala Leu Asn Gln Gly Tyr Ser Gly Thr
1280 1285 1290
Leu Gly Lys His Thr Ser Gly Leu Val Ala Arg Met Met Gly Ala
1295 1300 1305
Lys Met Pro Gly Gly Phe Gly Leu Ser Ala Ala Lys Ala His Leu
1310 1315 1320
Ser Lys Ala His Gly Leu Gly Pro Gly Arg Thr Asp Gly Ala Leu
1325 1330 1335
Leu Val Ala Leu Thr Lys Glu Pro Glu Lys Arg Leu Gly Ser Glu
1340 1345 1350
Ala Asp Ala Lys Ala Trp Leu Asp Gly Val Ala Gln Ala Tyr Ala
1355 1360 1365
Ser Gln Ala Gly Ile Thr Leu Gly Ala Gly Gly Gly Gly Gly Gly
1370 1375 1380
Ala Ala Val Gly Gly Ala Gly Phe Met Ile Asn Thr Glu Gln Leu
1385 1390 1395
Asp Lys Met Gln Glu Lys Gln Asp Asn Phe Val Ser Gln Gln Val
1400 1405 1410
Glu Leu Phe Leu Arg Tyr Leu Gly Lys Asp Ser Arg Glu Gly His
1415 1420 1425
Arg Leu Ala Asp Met Gln Lys Ala Glu Val Ala Asn Leu Gln Glu
1430 1435 1440
Lys Leu Asp Ser Ile Ala Arg Glu His Gly Asp Ala Tyr Val Gln
1445 1450 1455
Gly Ile Gln Pro Val Phe Asp Pro Leu Lys Ala Arg His Phe Asn
1460 1465 1470
Ser Ser Trp Asn Trp Val Arg Gln Asp Ala Leu Met Met Trp Met
1475 1480 1485
Asp Ile Leu Phe Gly Arg Leu Thr Thr Val Asp Arg Asp Ile Thr
1490 1495 1500
Ala Arg Cys Leu Val Ile Met Asn Arg Ala Asp Pro Ser Leu Ile
1505 1510 1515
Asp Tyr Met Gln Tyr Thr Ile Asp Asn Thr Pro Val Glu Arg Gly
1520 1525 1530
Glu His Tyr Val Leu Ala Lys Gln Phe Gly Gln Gln Leu Leu Asp
1535 1540 1545
Asn Cys Arg Glu Met Ile Gly Gln Ala Pro Leu Tyr Lys Asp Val
1550 1555 1560
Thr Phe Pro Thr Ala Pro Lys Thr Thr Val Asn Ala Lys Gly Asp
1565 1570 1575
Ile Ile Thr Glu Glu Val Asn Arg Pro Gly Val Ser Arg Leu Glu
1580 1585 1590
Lys Tyr Val Ala Glu Met Ala Ala Gly Ser Lys Val Thr Val Ala
1595 1600 1605
Ser Val Asn Leu Asp Lys Val Gln Glu Gln Val Glu Lys Leu Tyr
1610 1615 1620
Lys Leu Val Lys Ser Gln Pro Gln Ile Ser Lys Gln His Met Thr
1625 1630 1635
Ser Ile Lys Ser Leu Tyr Ala Glu Val Val Arg Gly Leu Gly Lys
1640 1645 1650
Asp Ala Gly Pro Pro Pro Val His Lys Ala Gly Thr Arg Ala Arg
1655 1660 1665
Arg Pro Ser Ser Gln Phe Leu Arg Pro Ala Ala Val Ser Glu Ala
1670 1675 1680
Thr Phe Leu Pro Glu Asp Lys Val Pro Leu Leu His Leu Lys Arg
1685 1690 1695
Lys Ile Gly Asn Asp Trp Gln Tyr Ser Ser Lys Leu Thr Ser Leu
1700 1705 1710
Tyr Leu Asp Ile Leu Lys Glu Ile Ala Thr Ser Gly Val Thr Phe
1715 1720 1725
Glu His Lys Asn Ala Leu Met Thr Gly Val Gly Lys Gly Ser Ile
1730 1735 1740
Gly Ile Glu Ile Val Lys Gly Leu Leu Ala Gly Gly Ala Arg Val
1745 1750 1755
Val Ile Thr Thr Ser Arg Tyr Ser Arg Ser Thr Val Glu Tyr Tyr
1760 1765 1770
Gln Ala Ile Tyr Gln Glu Val Gly Ser Lys Gly Ser Ser Leu Thr
1775 1780 1785
Val Val Pro Phe Asn Gln Gly Ser Lys Gln Asp Val Glu Ala Leu
1790 1795 1800
Val Asp Phe Ile Tyr Ser Lys Asp Lys Gly Leu Gly Met Asp Leu
1805 1810 1815
Asp Tyr Ile Leu Pro Phe Ala Ala Leu Pro Glu Asn Gly Arg Glu
1820 1825 1830
Ile Asp Gly Ile Asp Asp Arg Ser Glu Leu Ala His Arg Ile Met
1835 1840 1845
Leu Thr Asn Leu Leu Arg Leu Leu Gly Ala Val Lys Ser Lys Lys
1850 1855 1860
Ala Ala Leu Lys Leu Thr Thr Arg Pro Thr Glu Val Val Leu Pro
1865 1870 1875
Leu Ser Pro Asn His Gly Leu Phe Gly Asn Asp Gly Leu Tyr Ser
1880 1885 1890
Glu Ser Lys Ile Ser Leu Glu Thr Leu Phe Asn Arg Trp Ser Ser
1895 1900 1905
Glu Ser Trp Gly Glu Tyr Leu Cys Leu Ala Gly Ala Val Ile Gly
1910 1915 1920
Trp Thr Arg Gly Thr Gly Leu Met Ser Ala Thr Asn Ser Val Ala
1925 1930 1935
Glu Gly Ile Glu Ala Gln Gly Cys Arg Thr Phe Ser Ala Lys Glu
1940 1945 1950
Met Ala Phe Asn Ile Leu Gly Leu Met His Pro Leu Val Phe Asp
1955 1960 1965
Val Ala Gln Ile Glu Pro Val Trp Ala Asp Leu Asn Gly Gly Met
1970 1975 1980
Asp Lys Leu Pro Asp Leu Ala Asn Leu Thr Thr Glu Ile Arg Lys
1985 1990 1995
Lys Leu Asn Leu Thr Ala Ser Thr Arg Arg Ala Ile Ala Lys Asp
2000 2005 2010
Asn Ser Phe Asp Tyr Lys Val Ala His Gly Pro Ala Met Glu Gln
2015 2020 2025
Ile His Gln Arg Ile Asn Val Ala Pro Arg Ala Asn Phe Ser Leu
2030 2035 2040
Pro Phe Pro Glu Leu Lys Pro Ile Asp Ala Lys Ser Glu Leu Ala
2045 2050 2055
Lys Leu Arg Gly Leu Ile Asp Leu Glu Lys Val Val Val Met Thr
2060 2065 2070
Gly Tyr Ala Glu Val Gly Pro Phe Gly Ser Ser Arg Thr Arg Trp
2075 2080 2085
Glu Met Glu Ala Asn Gly Thr Phe Ser Ile Gln Gly Thr Leu Glu
2090 2095 2100
Leu Ala Tyr Val Met Gly Leu Ile Lys His Phe Glu Gly Arg Leu
2105 2110 2115
Lys Asp Gly Thr Leu Tyr Val Gly Trp Val Asp Ala Lys Thr Asn
2120 2125 2130
Glu Pro Leu Asp Asp Lys Asp Val Lys Ala Ala Tyr Glu Lys His
2135 2140 2145
Ile Leu Ala His Thr Gly Ile Arg Leu Ile Glu Pro Glu Ile Phe
2150 2155 2160
Asn Gly Tyr Asp Pro Lys Arg Lys Gly Phe Thr Gln Glu Ile Glu
2165 2170 2175
Ile Gln His Asp Leu Glu Pro Ile Glu Ala Ser Glu Glu Asp Ala
2180 2185 2190
Ala Arg Phe Lys Arg Glu His Gly Ala Leu Val Asp Val Tyr Thr
2195 2200 2205
Glu Asp Gly Ser Lys Phe Phe Val Lys Phe Lys Lys Gly Ala Lys
2210 2215 2220
Leu His Ile Pro Lys Ala Val Ala Phe Asp Arg Leu Val Ala Gly
2225 2230 2235
Gln Ile Pro Thr Gly Trp Ser His Lys Ala Phe Gly Ile Pro Asp
2240 2245 2250
Asp Ile Ala Ser Gln Val Asp Arg Thr Ser Leu Trp Ala Leu Val
2255 2260 2265
Ser Val Ala Glu Ala Leu Met Met Ala Gly Ile Thr Asp Pro Tyr
2270 2275 2280
Glu Leu Tyr Lys Trp Ile His Pro Ser Glu Val Gly Ser Ser Leu
2285 2290 2295
Gly Ser Gly Met Gly Gly Ile Thr Ser Ile Ser Lys Met Phe Arg
2300 2305 2310
Asp Arg Arg Glu Glu Lys Asp Val Gln Lys Asp Ile Leu Gln Glu
2315 2320 2325
Thr Phe Ile Asn Thr Val Ala Gly Trp Val Asn Leu Leu Leu Leu
2330 2335 2340
Ser Ser Ser Gly Pro Ile Lys Ile Pro Val Gly Ala Cys Ala Thr
2345 2350 2355
Ala Leu Gln Ser Val Glu Ile Ala Cys Asp Thr Ile Leu Ser Gly
2360 2365 2370
Lys Ala Lys Ile Met Val Ser Gly Gly Tyr Asp Asp Phe Ser Glu
2375 2380 2385
Glu Gly Ser Tyr Glu Phe Ala Asn Met Lys Ala Thr Ser Asn Ser
2390 2395 2400
Glu Thr Glu Phe Ala Ala Gly Arg Glu Pro Asn Glu Met Ser Arg
2405 2410 2415
Pro Thr Thr Ser Thr Arg Ala Gly Phe Met Glu Ser Met Gly Cys
2420 2425 2430
Gly Ala Gln Val Leu Met Ser Ala Lys Thr Ala Ile Glu Met Gly
2435 2440 2445
Ala Thr Ile Tyr Gly Ile Val Ala Tyr Thr Ala Thr Ala Thr Asp
2450 2455 2460
Lys Ala Gly Arg Ser Ile Pro Ala Pro Gly Arg Gly Val Met Gly
2465 2470 2475
Thr Ala Arg Glu Ile Thr Ser Lys Tyr Pro Ser Pro Ile Leu Asp
2480 2485 2490
Val Thr Tyr Arg Arg Arg Gln Leu Glu Phe Arg Arg Lys Gln Ile
2495 2500 2505
Ser Gln Trp Leu Glu Asn Glu Thr Glu Leu Leu Lys Phe Glu Val
2510 2515 2520
Ser Ser His Gly Gln Ala Thr Lys Leu Pro Asp Asp Tyr Val Ser
2525 2530 2535
Glu Arg Leu Ala Ser Ile Glu Arg Glu Ala Lys Arg Gln Glu Ala
2540 2545 2550
Glu Ala Leu Ala Thr Tyr Gly Met Leu Ala Gly Gln Asp Pro Thr
2555 2560 2565
Ile Ala Pro Leu Arg Arg Ala Leu Ala Val Trp Gly Leu Thr Ile
2570 2575 2580
Asp Asp Val Gly Val Ala Ser Phe His Gly Thr Ser Thr Val Ala
2585 2590 2595
Asn Asp Lys Asn Glu Ser Asn Ala Tyr Asn Glu Gln Phe Arg His
2600 2605 2610
Leu Gly Arg Ala Lys Gly Asn Ala Cys Pro Val Ile Ala Gln Lys
2615 2620 2625
Trp Leu Thr Gly His Pro Lys Gly Gly Ala Ala Ala Trp Met Leu
2630 2635 2640
Asn Gly Leu Ala Gln Val Ile Gln Ser Gly Leu Val Pro Gly Asn
2645 2650 2655
Arg Asn Ala Asp Asn Ile Gly Glu Glu Leu Arg Ala Phe Glu Tyr
2660 2665 2670
Leu Leu Tyr Pro Ser Lys Ser Ile Gln Thr Asp Gly Ile Lys Ala
2675 2680 2685
Gly Leu Leu Thr Ser Phe Gly Phe Gly Gln Val Gly Gly Gln Ala
2690 2695 2700
Leu Ile Val His Pro Ser Leu Leu Ile Gly Ala Leu Glu Pro Ala
2705 2710 2715
Gln Phe Glu Ala Tyr Lys Lys Leu Asn Asp Gln Arg Lys Lys Trp
2720 2725 2730
Ser Tyr Arg Arg Phe Asn Asp Phe Phe Thr Asn Gly Lys Leu Val
2735 2740 2745
Ile Ile Lys Asp Gly Thr Pro Phe Thr Pro Glu Gln Glu Asn Thr
2750 2755 2760
Thr Leu Leu Asn Pro Leu Val Arg Ala Val Pro Asp Lys Thr Gly
2765 2770 2775
Ser Tyr Ser Met Pro Lys Glu Phe Pro Ala Thr Val Pro Arg Ser
2780 2785 2790
Asn Asn Ala Glu Val Ala Asn Lys Leu Val Ser Ala Ala Val Gly
2795 2800 2805
Gly Ala Phe Gly Val Gly Thr Asp Val Glu Leu Ile Ser Ala Val
2810 2815 2820
Pro Thr Ser Glu Ser Phe Leu Glu Arg Asn Phe Thr Gln Asp Glu
2825 2830 2835
Ile Ala Tyr Cys Lys Ala Ala Pro Asp Phe Arg Ala Ser Leu Ala
2840 2845 2850
Ala Arg Trp Ser Ala Lys Glu Ala Thr Phe Lys Ala Leu Lys Thr
2855 2860 2865
Glu Ser Lys Gly Ala Ala Ala Ser Met Gln Asp Ile Glu Val Val
2870 2875 2880
Ser Thr Ser Gln Gly Pro Thr Ile Lys Leu His Gly Glu Val Glu
2885 2890 2895
Lys Ile Ala Gln Ala Ala Gly Ile Thr Ala Phe Glu Val Ser Leu
2900 2905 2910
Ser His Ser Glu Asp Val Ala Cys Ala Val Val Ile Ala Gln Lys
2915 2920 2925
<210> 10
<211> 4141
<212> PRT
<213> Aplanochytrium kerguelense
<400> 10
Met Glu Gln His Lys Thr Glu Lys Leu Ser Ala Ala Asp Glu Lys Leu
1 5 10 15
Ser Glu Ser Thr Ile His Trp Leu Ala Ala Thr Asp Val Pro Ser Asn
20 25 30
Phe Val Leu Phe Ser Gly Gln Gly Tyr Gln Tyr Phe Asp Glu Leu Arg
35 40 45
Glu Leu Tyr Glu Thr Gly Thr Asp Glu Val Lys Gly Phe Leu Leu Leu
50 55 60
Ala Ser Lys Thr Leu Gln Glu Glu Ile Cys Ser Glu Lys Ala Lys Thr
65 70 75 80
Val Ala Phe Lys Phe Val Asn Glu Leu Asp Phe Thr Lys Trp Ile Glu
85 90 95
Glu Asp Ile Asp Val Asp Gln Thr Tyr Ile Phe Ser Ala Pro Val Ser
100 105 110
Phe Pro Leu Ile Phe Val Ala Gln Val Ala Arg Tyr Leu Gln Thr Leu
115 120 125
Asn Leu Leu Arg Thr Asp His Lys Gln Phe Ile Gln Thr Ile Lys Gly
130 135 140
Ser Ser Gly His Ser Gln Gly Ile Val Ala Ala Val Leu Ile Ser Thr
145 150 155 160
Ser Pro Asp Asn Asp Arg Ile Val Glu Asn Ala Val Lys Leu Cys Arg
165 170 175
Tyr Met Leu Trp Gln Gly Leu Arg Cys His Glu Ser Ser Val Asn Arg
180 185 190
Phe Ser Gly Lys Asn His Lys Ser Lys Lys Ser Lys Val Tyr Ser Phe
195 200 205
Asp Glu Ser Thr Arg Met Val Lys Tyr Ser Pro Met Leu Ala Val Asn
210 215 220
Gly Leu Thr Glu Ser Leu Leu Asn Asp Phe Met Lys Lys Thr Glu Ser
225 230 235 240
Ile Ile Glu Ala Arg Arg Thr Ile Gln Gln Lys Tyr Asn Arg Arg Glu
245 250 255
Ser Phe Phe Glu Thr Pro Thr Glu Lys Asp Glu Lys Ser Lys Arg Leu
260 265 270
Asn Met Phe Gln Ile Ala Leu Ala Asn Gly Pro Lys Ser Phe Val Val
275 280 285
Ser Gly Ala Pro Lys Thr Leu Ile Glu Leu Glu Gln Arg Ile Lys Glu
290 295 300
Ser Thr Thr Ser Ser Pro Asn Ser Gln Thr Arg Ile Pro Phe Ser Lys
305 310 315 320
Arg Lys Pro Val Val Lys Met Tyr Phe Leu Lys Val Gly Ala Ala Phe
325 330 335
His Thr Glu Ile Cys Asn Asp Ala Ser Ser Lys Leu Lys Ala Asp Val
340 345 350
Val Gln Met Lys Leu Ser Ile Ser Pro Lys Glu Ile Leu Ile Pro Val
355 360 365
Tyr His Thr Lys Asp Gly Ser Asn Leu Ser Glu Ile Ser Ala Asp Val
370 375 380
Asn Val Val Asp Leu Leu Ile Asp Met Gln Thr Ser Gln Val Asn Asp
385 390 395 400
Phe Arg Lys Thr Leu Lys Ser Ile Ser Ser Gln Asn Gly Val Ser Asn
405 410 415
Val Ile Asp Phe Gly Pro Gly Asp Gly Thr Ala Lys Leu Cys Leu Lys
420 425 430
Arg Lys Ser Gly Thr Gly Ile Leu Val Val Ala Ala Thr Gly Pro Leu
435 440 445
Arg Asn Arg Arg Cys Ala Tyr Gly Arg Asn Leu Val Gly Leu Asn Ser
450 455 460
Val Leu Leu Glu Lys Asn Pro Ile Leu Gly Met Asn Trp Gly Glu Glu
465 470 475 480
Phe Lys Pro Arg Leu Ser Ser Arg Lys Asn Asp Asn Gln Ile Ile Val
485 490 495
Gly Thr Arg Phe Ser Asp Leu Thr Gly Lys Pro Pro Val Ile Leu Pro
500 505 510
Gly Met Thr Pro Thr Thr Ser Phe His Gly Ile Asp Leu Val Ala Ala
515 520 525
Cys Ser Asn Gly Gly Tyr His Gly Glu Leu Ala Gly Gly Gly Leu Pro
530 535 540
Leu Pro Glu Tyr Phe Lys Ala Lys Ile Asp Glu Leu Val Ser Lys Gln
545 550 555 560
Asn Ser Gly Val Gly Ile Asn Ile Asn Met Leu Tyr Leu Asn Ser Tyr
565 570 575
Leu Trp Gly Phe Gln Phe Ser Thr Ala Ile Gln Met Ala Lys Asp Gly
580 585 590
Tyr Pro Ile Glu Ser Ile Thr Cys Ala Ala Gly Val Pro Thr Thr Glu
595 600 605
Lys Ala Lys Glu Ile Met His Gln Leu Lys Glu Ala Gly Ile Lys Tyr
610 615 620
Ile Ala Phe Lys Pro Gly Ser Arg Ala Ala Ile Met Asp Val Leu Ser
625 630 635 640
Ile Ala Arg Glu Asn Pro Asn Ser Ser Ile Val Leu Gln Trp Thr Gly
645 650 655
Gly Arg Gly Gly Gly His His Ser Phe Glu Asp Phe His Ala Pro Leu
660 665 670
Leu Asp Thr Tyr Gln Glu Ile Arg Glu His Pro Asn Val Val Leu Val
675 680 685
Val Gly Ser Gly Phe Gly Asp Ala Glu Lys Ser Tyr Pro Tyr Leu Asp
690 695 700
Gly Ser Trp Ser Thr Leu Pro Pro His Asn Cys Glu Ser Arg Met Pro
705 710 715 720
Cys Asp Ala Val Leu Phe Gly Ser Arg Cys Met Val Ala Lys Glu Ala
725 730 735
Ala Thr Ser Pro Glu Val Lys Gln Leu Ile Val Asn Ala Lys Gly Val
740 745 750
Arg Glu Asn Glu Ser Trp Glu Met Ser Tyr Glu Asn Asp Ala Gly Gly
755 760 765
Ile Leu Thr Val Thr Ser Glu Leu Gly Glu Pro Ile His Lys Ile Asn
770 775 780
Asn Arg Gly Met Lys Leu Trp Arg Glu Phe Asp Arg Lys Tyr Phe Ser
785 790 795 800
Leu Pro Ser Gly Gln Glu Arg Glu Lys Ala Ile Val Lys Asp Lys Lys
805 810 815
Glu Ile Ile Arg Arg Ile Asn Ala Asp Phe Gln Lys Val Tyr Phe Gly
820 825 830
Arg Lys Ala Asp Gly Thr Val Val Asp Ile Ala Tyr Met Thr Tyr Gly
835 840 845
Glu Ile Leu Arg Arg Met Val Asp Leu Met Tyr Val Thr Gly Gly Gly
850 855 860
Asp Gly Lys Gln His Arg Phe Ala Pro Asn Arg Trp Ile Asp Val Thr
865 870 875 880
Tyr Gln Thr Arg Thr Phe Lys Phe Leu Val Arg Thr Glu Arg Arg Phe
885 890 895
Leu Arg Asp Lys Lys Met Ala Phe Ala Thr Asp Met Arg Lys Leu Glu
900 905 910
Ser Phe Pro Ile Lys Cys Ile Asp Glu Phe Val Lys Cys Tyr Pro Gln
915 920 925
Val Asp Lys Val Leu Val Ser Asp Glu Asp Ala Gly Tyr Phe Ile Glu
930 935 940
Leu Cys Arg Asn Leu Arg Asn Gly Lys Pro Val Asn Phe Ile Pro Gln
945 950 955 960
Ile Asp Gly Ser Leu Ala Tyr Trp Phe Lys Lys Asp Ser Leu Trp Cys
965 970 975
Ser Glu Gln Ile Asp Ala Val Pro Asp Asn Asp Pro Gly Arg Val Cys
980 985 990
Ile Leu His Gly Pro Val Ala Ala Gln Tyr Ser Val Val Ser Asp Glu
995 1000 1005
Pro Ile Cys Glu Ile Leu Gly Asn Ile His Lys Gly Tyr Val Asp
1010 1015 1020
Lys Leu Lys Glu Glu Lys Tyr Glu Val Ala Lys Val Glu Thr Ile
1025 1030 1035
Gly Lys Leu Pro Glu Tyr Lys Gly Ser Asn Lys Ser Phe Arg Lys
1040 1045 1050
Phe Leu Arg Tyr Gly Trp Phe Asn Ala Leu Phe Gln Ser Thr Val
1055 1060 1065
Met Val Lys Glu Lys Lys Trp Val Arg Asn Ile Ile Pro Gly Ile
1070 1075 1080
Ile Gln Thr Glu Asn Glu Asn Ile Val Phe Ser Glu Arg Thr Ala
1085 1090 1095
Met Asn Asp Met Cys Leu Ile Glu Val Leu Leu Asn Asp Ser Gln
1100 1105 1110
Arg Thr Leu Ala Phe Ile Glu Phe Glu Gly Lys His Ile Ser Val
1115 1120 1125
Ser Leu Gln Asp Ser Glu Pro Val Asn Ser Thr Leu Thr Leu Tyr
1130 1135 1140
Phe Glu Tyr His Pro Glu Thr Pro His Ser Pro Leu Phe Gln Val
1145 1150 1155
Thr Asn Gly Ser Lys Ser Ile Ala Arg Ser Ile Lys Gln Tyr Tyr
1160 1165 1170
Arg Asn Ile Trp Asp Cys Gln Asp Val Val Ser Ile His Asp Thr
1175 1180 1185
Phe Ser Glu Ser Phe Thr Val Thr Arg Glu Asp Ile Glu Arg Phe
1190 1195 1200
Asn Gly Ala Ile Gln Tyr Ala Val Asn Thr Ser Glu Gly Thr Leu
1205 1210 1215
Asp Phe Ala Ile Val Ala Gly Trp Lys Ser Leu Ile Lys Ala Leu
1220 1225 1230
Phe Ser Lys Glu Ile Asp Gly Ser Leu Leu Asp Leu Val His Ile
1235 1240 1245
Ser His Ser Tyr Lys Leu Leu Val Glu Glu Lys Glu Arg Thr Leu
1250 1255 1260
Val Gln Ala Gly Glu Lys Ile Cys Ser Glu Ser Arg Val Ile Ser
1265 1270 1275
Val Gln Ile Leu Pro Gly Val Gly Lys Ser Ile Thr Val Lys Gly
1280 1285 1290
Val Leu Ser Arg Asn Thr Val Lys Trp Val Glu Val Gln Ser Glu
1295 1300 1305
Phe Leu Ile Arg Gly Asn Phe Phe Asp Phe Gln Lys Thr Phe Lys
1310 1315 1320
Glu Tyr Ser Tyr Lys Ala Glu Val Leu Cys Lys Asp Asp Thr Val
1325 1330 1335
Pro Gln Ile Leu Asn Ser Gln Glu Trp Leu Phe Leu Asp Ser Gly
1340 1345 1350
Val Arg Pro Leu Ser Lys His Asp Lys Val Val Phe Gln Ile Asp
1355 1360 1365
Arg Val Val Glu Lys Arg Lys Ser Val Thr Asn Ile Ser Asp Ile
1370 1375 1380
Leu Val Ala Gly Asn Val Val Arg Ile Asn Thr Pro Glu Glu Ile
1385 1390 1395
Asp Gly Phe Ile Glu Leu Lys Asp Gln Gly Ser Pro Lys Gly Ile
1400 1405 1410
Ile Ile Gly Lys Val Asp Leu Ser Leu Glu Asn Ala Asn Leu Ser
1415 1420 1425
Glu Asn Pro Ile Ile Thr Tyr Leu Asp Ser Ile Lys Glu Ser Lys
1430 1435 1440
Leu His Gly Ser Val Phe Glu Ser Gly Gly Tyr Thr Val Met Pro
1445 1450 1455
Arg Pro Glu Leu Val Thr Ala Pro Ser Ile Val Glu Ser Ile Val
1460 1465 1470
Tyr Ala Ser Ala Ser Arg Asp Met Asn Pro Ile His Arg Asn Arg
1475 1480 1485
Thr Phe Ala Lys Leu Ala Gly Leu Pro Gly Gly Ser Thr Ile Val
1490 1495 1500
His Gly Met Trp Thr Ala Ala Met Ser Arg Arg Ile Leu Glu Leu
1505 1510 1515
His Thr Val Leu Gly Asp His Arg Arg Ile Ser Thr Tyr Ser Val
1520 1525 1530
Lys Phe Thr Gly Met Val Phe Pro Glu Asp Lys Leu Ala Val Met
1535 1540 1545
Val Lys His Val Gly Val Thr Glu Gly Arg Leu Ile Leu Asp Val
1550 1555 1560
Asp Val Ser Lys Val Glu Thr Gln Glu Arg Val Leu His Gly Arg
1565 1570 1575
Ala Glu Val Glu Gly Pro Ser Thr Ser Phe Leu Phe Thr Gly Gln
1580 1585 1590
Gly Ser Ala Lys Val Gly Met Gly Met Asp Ser Tyr Glu Glu Lys
1595 1600 1605
Gln Ser Ala Arg Ala Val Trp Gln Thr Ala Asp Lys Phe Leu Arg
1610 1615 1620
Lys Lys Phe Gly Phe Ser Ile Val Asp Ile Val Lys Asn Asn Pro
1625 1630 1635
Lys Glu Leu Thr Val Arg Phe Gly Gly Ser Gln Gly Lys Val Ile
1640 1645 1650
Arg Glu Asn Tyr Arg Ala Ile Lys Arg Glu Asp Gly Ser Gln Leu
1655 1660 1665
Ile Arg Glu Ile His Ser Asn Thr Thr Ser Phe Thr Phe Arg Ser
1670 1675 1680
Pro Asn Gly Leu Leu Phe Ala Thr Gln Phe Ser Gln Pro Ala Leu
1685 1690 1695
Val Leu Val Gln Lys Ala Ala Phe Glu Glu Met Leu Gln Gly Gly
1700 1705 1710
Phe Val Pro Ser Asn Ser Met Phe Ala Gly His Ser Leu Gly Glu
1715 1720 1725
Tyr Ala Ala Leu Ala Ser Tyr Ala Asn Val Leu Ser Ile Glu Asp
1730 1735 1740
Leu Val Glu Thr Val Phe Leu Arg Gly Met Val Met Gln Asn Ala
1745 1750 1755
Val Ala Arg Asp Asp Glu Gly Thr Ser Asp Tyr Gly Met Val Ala
1760 1765 1770
Ala Asn Pro Ser Arg Val Ser Lys Thr Phe Ser Ala Gln Leu Leu
1775 1780 1785
Met Glu Ile Val Gln Lys Ile Asp Asp Glu Ser Asp Asp Ser His
1790 1795 1800
Leu Leu Gln Val Val Asn Phe Asn Ile Arg Asp Ala Gln Tyr Val
1805 1810 1815
Ile Ala Gly Asn Leu Glu Leu Leu Asp Ser Leu Ser Asn Val Leu
1820 1825 1830
Asn Val Ile Ser Asn Ser His Asp Lys Thr Phe Asp Arg Ser Ile
1835 1840 1845
Ile Gln Ala Ala Met Ile Lys Ala Glu Asn Arg Arg Arg Ser Cys
1850 1855 1860
Lys Glu Asn Asn Gln Pro Phe Arg Leu Lys Arg Gly Ile Ala Thr
1865 1870 1875
Ile Pro Leu His Gly Ile Asp Val Pro Phe His Ser Arg Gln Leu
1880 1885 1890
Leu Asn Gly Val Pro Ala Phe Arg Ser Leu Leu Glu Pro Arg Phe
1895 1900 1905
Thr Glu Glu Met Ile Gln Lys Tyr Leu Pro Arg Leu Ile Gly Lys
1910 1915 1920
Tyr Ile Pro Asn Val His Ala Glu Pro Phe Ser Ile Ser Lys Glu
1925 1930 1935
Tyr Ile Glu Lys Val Ala Ser Val Thr Lys Ser Pro Ala Leu Tyr
1940 1945 1950
Lys Leu Leu Ser Thr Tyr Ser Thr Met Cys Asp Ser Ser Lys Ala
1955 1960 1965
Arg Ile Leu Leu Ile Glu Leu Leu Ala His Gln Phe Ala Met Pro
1970 1975 1980
Val Gln Trp Ile Asp Thr Gln Asp Tyr Ile Leu Ser Arg His Thr
1985 1990 1995
Gln Arg Ile Ile Glu Met Gly Pro Ala Pro Thr Leu Val Gly Met
2000 2005 2010
Ala Lys Lys Thr Leu Gln Ser Arg Ile Tyr Gly Asp Glu Glu Asp
2015 2020 2025
Tyr Lys Pro Ser Ile Leu Trp Trp Asn Gln Asn Gln Glu Glu Ile
2030 2035 2040
Phe Tyr Asn Leu Asp Asn Gln Gly Ile Ser Ile Gln Gln Phe Leu
2045 2050 2055
Gln Glu Leu Lys Gly Thr Glu Asp Asp Glu Ser Gln Ser Asp Asp
2060 2065 2070
Met Ser Ser Tyr Thr Gln Ser Asp Glu Thr Ile Gln Arg Thr Ala
2075 2080 2085
Lys Asn Arg Val Met Pro Glu Ala Asn Ala Asn Asp Lys Glu Lys
2090 2095 2100
Val Gly Asn Glu Leu Gln Lys Lys Glu Val Thr Asn Arg Pro Leu
2105 2110 2115
His Val Leu Arg Val Leu Ile Ser Thr Lys Leu Lys Lys Lys Met
2120 2125 2130
Leu Glu Val Lys Ala Thr Asp Thr Ile Lys Gly Val Ser Gly Gly
2135 2140 2145
Lys Ser Ala Ile Gln Asn Glu Leu Ile Gly Glu Ile Asn Ala Glu
2150 2155 2160
Tyr Ser Ser Phe Arg Asp Ile Glu Asp Ile Ser Glu Val Ser Leu
2165 2170 2175
Ala Glu Leu Ala Arg Arg Ala Pro Ser Arg Gln Glu Thr Val Leu
2180 2185 2190
Gly Lys Val Thr Thr Lys Met Val Asn Lys Met Phe Ser Ser Lys
2195 2200 2205
Leu Pro Gly Lys Phe Gly Pro Asn Asp Thr Lys Glu Tyr Leu Gln
2210 2215 2220
Ser Lys Leu Ser Asp Ala Ser Thr Val Asp Ser Val Ser Leu His
2225 2230 2235
Ala Leu Thr Leu Gln Pro Val Ser Arg Leu Ala Thr Pro Gly Glu
2240 2245 2250
Ala Gln Ala Trp Leu Asp Ser Val Cys Arg Asp Phe Glu Asn Asp
2255 2260 2265
Gln Arg Ile Ser Ile Leu Lys Asn Lys Ser Pro Asn Ser Asn Glu
2270 2275 2280
Ser Val Met Ile Ser Glu Gln Arg Thr Val Gln Asn Asn Phe Val
2285 2290 2295
Gln Ile Ser Glu Glu Pro Val Asn Ser Lys His Val Leu Leu Ser
2300 2305 2310
Leu Ile Ser Met Lys Leu Gln Lys Glu Ile Ser Ser Ile Asn Glu
2315 2320 2325
Thr Ser Thr Ile Lys Ser Leu Val Asn Gly Lys Ser Ala Val Gln
2330 2335 2340
Asn Glu Ile Leu Gly Glu Ile Ala Ser Glu Phe Arg Thr Asp Glu
2345 2350 2355
Ser Glu Gly Ala Ala Asp Ile Ser Leu Ser Glu Leu Gly Glu Lys
2360 2365 2370
Leu Gly Ile Gly Tyr Ala Gly Pro Gly Ser Val Val Ser Arg His
2375 2380 2385
Ile Asn Lys Met Leu Asn Ser Lys Leu Pro Gly Ser Phe Asn Ser
2390 2395 2400
Ser Ser Ala Lys Lys Tyr Leu Ser Met Gln Gly Phe Gly Lys Gly
2405 2410 2415
Arg Ser Asp Ser Val Leu Val His Ala Leu Thr Met Gln Pro Lys
2420 2425 2430
Ser Arg Leu Gln Gly Val Glu Asp Ala Lys Ser Trp Leu Asp Ser
2435 2440 2445
Val Cys Ser Ser Tyr Gly Lys Trp Ala Asn Ile Asp Leu Thr Val
2450 2455 2460
Ser Arg Thr Leu Ser Ser Gly Asp Ser Ser Pro Ala His Ser Thr
2465 2470 2475
Lys Asp Ile Leu Ser Ser Thr Ala Met Lys Lys Leu Lys Lys Leu
2480 2485 2490
Tyr Leu Asp Gln Val Asp Val Phe Asn Thr Phe Leu Gly Glu Asp
2495 2500 2505
Arg Arg Ala Lys Leu Gln Lys Val Leu Thr Glu Glu Thr Ser Ala
2510 2515 2520
Lys Asp Arg Ile Glu Ala Gln Leu Leu Glu Leu Lys Glu Glu Leu
2525 2530 2535
Gly Asp Pro Phe Cys Asp Gly Ile Gln Pro Gln Phe Asn Glu Ala
2540 2545 2550
Arg Ile Arg Ile Tyr Asp Ser Tyr Trp Asn Trp Val Val Gln Asp
2555 2560 2565
Ala Leu Glu Leu His Tyr His Thr Phe Ser Cys Val Leu Asn Ser
2570 2575 2580
Lys Gly Lys Asn Val Asn Ile Pro Asn Ser Ser Asn Ser Tyr Phe
2585 2590 2595
Arg Ala Met Ser Asp Trp Ile Thr Ser Ser Asn Glu Lys Leu Glu
2600 2605 2610
Glu Asn Lys Pro Pro Gln Ala Trp Phe Arg Asn Tyr Leu Cys Asn
2615 2620 2625
Arg Ala Thr Pro Glu Leu Leu Thr Val Val Gln Tyr Phe Ala Ala
2630 2635 2640
Arg Met Asn Glu Gln Gly His Ser Glu Tyr Ala Gln Ala Val Ser
2645 2650 2655
Leu Leu Ala Glu Gln Val Ser Gln Trp Ile Ser Arg Pro Pro Val
2660 2665 2670
His Ile Ala Leu Phe Ala Ser Leu Glu Pro Arg Val Thr Val Asp
2675 2680 2685
Ser Ala Asn Asn Phe Gly Leu Lys Tyr Glu Glu Lys Pro Arg Lys
2690 2695 2700
Phe Val Ala Val Gly Glu Asn Lys Ala Phe Ser Cys Asp Asn Ala
2705 2710 2715
Ser Leu Tyr Val Lys Glu Met Ser Lys Gly Leu Phe Tyr Asp His
2720 2725 2730
Arg Val Ala Ser Lys Val Glu His Pro Ser Gln Ser Val Tyr Leu
2735 2740 2745
Ser Pro Asp Asn Ser Phe Phe Pro Asn Glu Glu Glu Ser Asp Gln
2750 2755 2760
Ile Ser Ser Gly Met Arg Leu Pro Lys Thr Gln Gly Glu Ile Gln
2765 2770 2775
Arg Asp Phe Ser Arg Leu Pro Ala Gly Gln Lys Leu Glu Val Met
2780 2785 2790
Arg Lys Ser Val Asn Arg Ser Ser Asp Thr Asp Glu Glu Ile Ala
2795 2800 2805
Asn Ile Val Arg Gln Asn Tyr Asn Ser Ile His Val Ala Lys Asn
2810 2815 2820
Val Pro Leu Val His Leu Lys Ser Pro Ser Lys Phe Asp Lys Thr
2825 2830 2835
Val Arg Val Leu Asp Glu Pro Leu Thr Ser Met Tyr Leu Ser Cys
2840 2845 2850
Leu His Asp Ile Ala Thr Ser Gly Val Ser Phe Ala Gly Gln Asn
2855 2860 2865
Ala Leu Val Thr Gly Ala Gly Phe Gly Ser Ile Gly Ile Glu Leu
2870 2875 2880
Ile Lys Pro Leu Leu Glu Gly Gly Ala Thr Val Leu Val Thr Val
2885 2890 2895
Arg Leu Asn Arg Thr Asp Glu Gln Met Gln Ile Val Asn Glu Arg
2900 2905 2910
Phe Gln Arg Leu Tyr Glu Glu Phe Gly Ser Arg Gly Ser Lys Leu
2915 2920 2925
Val Leu Val Pro Cys Asn Cys Ala Ser Asn Gln Asp Val His Ser
2930 2935 2940
Leu Ile Arg His Ile Tyr Glu Lys Leu Lys Leu Asp Leu Asp Phe
2945 2950 2955
Ile Phe Pro Phe Ala Ala Ile Gly Glu Gln Gly Lys Asp Val Ser
2960 2965 2970
Glu Ile Gly Ser Lys Ser Glu Val Ala His Arg Leu Met Leu Thr
2975 2980 2985
Asn Thr Ile Arg Leu Leu Gly Ala Val Lys Lys Ala Lys Glu Asp
2990 2995 3000
Arg Cys Ile Glu Thr Arg Pro Ala Leu Val Leu Leu Pro Cys Ser
3005 3010 3015
Pro Asn His Gly Asp Phe Gly Leu Asp Gly Leu Tyr Ala Glu Ser
3020 3025 3030
Lys Leu Gly Leu Glu Ser Leu Val Asn Lys Trp Lys Ser Glu Glu
3035 3040 3045
Trp Gly Asn Tyr Leu Ser Ile Cys Ala Ala Val Ile Gly Trp Thr
3050 3055 3060
Arg Ser Lys Leu Met Trp Gln Asn Asn Val Val Ala Glu Gly Ile
3065 3070 3075
Glu Lys Leu Gly Val Arg Thr Phe Ser Thr Thr Glu Thr Ala Phe
3080 3085 3090
Asn Leu Ile Gly Leu Leu His Pro Asp Ile Val Ser His Ala Ala
3095 3100 3105
Glu Glu Pro Leu Trp Ala Asp Leu Thr Gly Asn Trp Gly Ala Ala
3110 3115 3120
Pro Asp Leu Lys Glu His Ser Lys Arg Ile Arg Thr Ser Leu Leu
3125 3130 3135
Ala Gln Ser Lys Ala Ala Lys Ala Ile Ser Leu Ser Ser Lys Ser
3140 3145 3150
Leu Val Ile Glu Pro Ser Glu Ser Lys Thr Lys Ala Gln Thr Glu
3155 3160 3165
Ile Val Asn Thr Tyr Leu Ala Gly Val Asn Asp Lys Leu Arg Leu
3170 3175 3180
Pro Leu Ala Asn Pro Glu Lys Phe Cys Asn Pro Phe Pro Lys Ile
3185 3190 3195
Pro Ser Gln Glu Arg Met Asp Ser Leu Ala Tyr Leu Lys His Ser
3200 3205 3210
Val Asp Leu Lys Lys Val Val Val Val Val Gly Tyr Gly Glu Ile
3215 3220 3225
Gly Pro Trp Gly Asn Ser Arg Thr Arg Trp Glu Met Glu Ser Phe
3230 3235 3240
Gly Glu Phe Ser Leu Glu Gly Ala Ile Glu Leu Ala Trp Leu Val
3245 3250 3255
Gly Leu Ile Lys Pro Val Thr Gly Pro Leu Lys Asn Asp Pro Arg
3260 3265 3270
Thr Gln Tyr Phe Gly Trp Val Asp Ala Glu Ser Glu Glu Pro Val
3275 3280 3285
Ala Asp His Glu Ile Lys Thr Arg Tyr Glu Lys Val Leu Leu Gln
3290 3295 3300
His Ser Gly Ile Arg Leu Ile Glu Pro Glu Leu Phe Glu Gly Tyr
3305 3310 3315
Asn Pro Lys Lys Lys Ser Ile Leu Arg Gln Val Ala Ile Ala Glu
3320 3325 3330
Asp Met Lys Pro Ile Glu Val Ala Ser Leu Glu Glu Ala Gln Gln
3335 3340 3345
Tyr Val Asn Glu Leu Gly Lys Glu Phe Ile Asp Val Phe Asn Glu
3350 3355 3360
Asn Val Glu Ser Asn Asp Gly Gln Trp Tyr Ile Arg Leu Lys Ala
3365 3370 3375
Gly Ala Val Val Ser Ile Pro Gly Ala Leu Ser Phe Asn Arg Phe
3380 3385 3390
Ile Ala Gly Gln Leu Pro Thr Gly Trp Asp Ala Lys Arg Leu Gly
3395 3400 3405
Ile Pro Asp Asp Ile Ala Asp Ser Val Asp Pro Val Thr Leu Tyr
3410 3415 3420
Ala Leu Val Ser Thr Val Glu Ala Leu Val Cys Ala Gly Leu Thr
3425 3430 3435
Asp Pro Tyr Glu Leu Tyr Gln Tyr Val His Val Ser Gln Val Gly
3440 3445 3450
Asn Thr Ser Gly Gly Gly Met Gly Gly Met Arg Ser Leu Lys Arg
3455 3460 3465
Met Phe Leu Glu Arg Lys Leu Asp Ala Glu Ile Pro Ser Asp Thr
3470 3475 3480
Leu Ala Glu Ser Phe Ile Asn Thr Met Pro Ala Trp Val Asn Met
3485 3490 3495
Leu Leu Leu Ser Ser Ser Gly Pro Ile Lys Thr Pro Val Gly Ala
3500 3505 3510
Cys Ala Thr Ala Ala Glu Ser Val Asp Ile Gly Ile Glu Thr Ile
3515 3520 3525
Leu Ala Gly Lys Ala Arg Val Val Ile Ala Gly Gly Tyr Asp Asp
3530 3535 3540
Phe Cys Glu Thr Gly Ser Asn Glu Phe Ala Met Met Gly Ala Thr
3545 3550 3555
Ser Asn Ser Gln Thr Glu Ala Glu Lys Gly Arg Phe Pro Arg Glu
3560 3565 3570
Ala Ser Arg Pro Met Thr Asp Thr Arg Ala Gly Phe Met Glu Ser
3575 3580 3585
Gln Gly Ala Gly Met Gln Val Leu Met Asp Ala Glu Leu Ala Ile
3590 3595 3600
Gln Met Gly Leu Pro Val Tyr Gly Ile Leu Ala Leu Ser Asn Thr
3605 3610 3615
Ala Thr Asp Arg Gln Gly Arg Ser Val Pro Ala Pro Gly Arg Gly
3620 3625 3630
Ile Leu Thr Thr Ala Arg Glu Val Arg Ser Lys Asn Ser Lys Ser
3635 3640 3645
Ser His Asn Gly Lys Pro Val Glu Asn Pro Leu Leu Ser Val Lys
3650 3655 3660
Phe Arg Lys Lys His Leu Arg Gln Glu Leu Asp Ala Ile Asp Leu
3665 3670 3675
Trp Ala Ser Gln Glu Ile Ser Asn Met Ser Glu Glu Phe Ser Gln
3680 3685 3690
Asn Ser Arg Lys Met Glu Asn Arg Lys Gly Phe Val Glu Thr Met
3695 3700 3705
Arg Lys Lys Lys Arg Ser Ala Ala Phe Glu Thr Trp Gly Gln Gly
3710 3715 3720
Phe Tyr Arg Asn Asp Asp Ser Ile Ala Pro Leu Arg Gly Ala Leu
3725 3730 3735
Ser Val Trp Gly Leu Thr Val Asp Asp Leu Leu Val Gly Ser Phe
3740 3745 3750
His Gly Thr Gly Thr Asn Leu Asn Asp Thr Asn Glu Ser Ser Leu
3755 3760 3765
Val Asn Lys Gln Leu Lys His Leu Asp Arg Lys Glu Gly Asn Ile
3770 3775 3780
Leu Leu Val Val Thr Gln Lys Tyr Leu Thr Gly His Pro Lys Gly
3785 3790 3795
Ala Ala Ala Ala Trp Met Leu Asn Gly Leu Leu Gln Cys Met Asn
3800 3805 3810
Ser Gly Arg Val Pro Gly Asn Arg Asn Leu Asp Asn Val Asp Gly
3815 3820 3825
Lys Leu Arg Thr Asn Gly Tyr Leu Phe Tyr Pro Asn Arg Thr Ile
3830 3835 3840
Glu Val Pro Arg Val Glu Ala Ala Phe Leu Lys Ser Phe Gly Phe
3845 3850 3855
Gly Gln Ala Gly Ala Glu Val Val Ile Ile His Pro Asp Arg Leu
3860 3865 3870
Leu Ala Val Leu Ser Glu Glu Asn Leu Lys Ser Tyr Ile Leu Arg
3875 3880 3885
Arg Asn Glu Arg Glu Lys Arg Ala Tyr Arg Tyr His Gln Gly Val
3890 3895 3900
Met Ser Gly His His Thr Met Val Gln Val Lys Glu Phe Ala Pro
3905 3910 3915
Tyr Glu Asp Asp Ile Leu Glu Glu Ile Tyr Leu Asn Pro Arg Ala
3920 3925 3930
Arg Ala Ser Phe Asp Ser Ser Lys Ser Thr Trp Thr Phe His Lys
3935 3940 3945
Tyr Arg Asn Asp Lys Asp Thr Val Asp Asp Lys Asp Glu Met Glu
3950 3955 3960
Gln Asp Ser Leu Asn Glu Ser Val Asp Leu Pro Lys Asp Ile Thr
3965 3970 3975
Lys Val Gly Ser Pro Ser Glu Ile Arg Ile Pro Val Lys Thr Arg
3980 3985 3990
Leu Glu Val Thr Val Arg Glu Gly Val Glu Gly Leu Thr Lys Lys
3995 4000 4005
Asp Lys Phe Ser Ser Gln Gly Val Gly Val Asp Val Glu Pro Val
4010 4015 4020
Ser Thr Phe Ala Gln His Glu Glu Lys Thr Ile Phe Ile Gln Asn
4025 4030 4035
Asn Phe Thr Glu Asn Glu Gln Leu Tyr Cys Asn His Ala Ala Ser
4040 4045 4050
Pro Ala Ala Ser Tyr Ala Gly Arg Trp Ala Ala Lys Glu Ala Val
4055 4060 4065
Ile Lys Ala Ile Ser Asn Ser Ser Leu Glu Thr Arg Ser Leu Trp
4070 4075 4080
Gln Gly Ala Glu Gly Lys Leu Ile Asp Ile Glu Ile Ile Gln Ser
4085 4090 4095
Asn Ser Gly Ala Pro Glu Val Val Leu His Gly His Ala Lys Glu
4100 4105 4110
Val Phe Gln Thr Leu Gly Leu Thr Asn Val Lys Val Ser Ile Ser
4115 4120 4125
His Thr Pro Glu Val Ala Val Ala Gln Ala Ile Thr Asn
4130 4135 4140
<210> 11
<211> 2944
<212> PRT
<213> 圆红冬孢酵母
<400> 11
Met Val Ala Ala Gln Asp Leu Pro Leu Ala Leu Ser Ile Ser Phe Ala
1 5 10 15
Pro Glu Ser Ser Thr Ile Ser Met Thr Leu Phe Asn Gln Pro Glu Ala
20 25 30
Ser Lys Pro Ala Leu Pro Leu Glu Leu Lys Tyr Lys Tyr Asp Pro Ser
35 40 45
Thr Pro Tyr Ala Pro Ile His Glu Ile Thr Glu Asp Arg Asn Gln Arg
50 55 60
Ile Lys Gln His Tyr Trp Asp Leu Trp Gly Leu Gly Asn Lys Ala Asp
65 70 75 80
Gln Gly Ile Ser Gln Leu Lys Ile Thr Asp Glu Phe Gln Gly Asp Leu
85 90 95
Val Thr Ile Ser Ala Asp Glu Ile Glu Ala Phe Cys Arg Val Val Gly
100 105 110
Ile Glu Gly Glu Ala Tyr Lys Arg Asn His Lys Ala Gly Met Gln Val
115 120 125
Pro Leu Asp Phe Ala Ile Lys Leu Gly Trp Lys Ala Ile Met Lys Pro
130 135 140
Ile Phe Pro Ser Thr Ile Asp Gly Asp Leu Leu Lys Leu Val His Leu
145 150 155 160
Ser Asn Gly Phe Arg Val Leu Pro Asp Thr Pro Thr Leu Gln Val Gly
165 170 175
Asp Val Val Thr Thr Thr Ser Arg Ile Glu Ser Ile Thr Asn Ser Asp
180 185 190
Thr Gly Lys Thr Val Ser Val Arg Gly Val Ile Ser Leu Val Ser Ser
195 200 205
Ala Asp Ser Lys Gly Lys Asp Ala Ser Thr Glu Asp Arg Ile Pro Leu
210 215 220
Ile Glu Val Thr Ser Ser Phe Phe Tyr Arg Gly Lys Phe Ser Asp Tyr
225 230 235 240
Ala Gln Thr Phe Ser Arg Val Ala His Pro Thr Tyr Ser Val Pro Ile
245 250 255
Thr Thr Pro Glu Ala Val Ala Val Leu Gln Ser Lys Glu Trp Phe Gln
260 265 270
Trp Asp Asp Asp Ser Lys Pro Leu Glu Val Gly Thr Lys Leu Gln Phe
275 280 285
Lys Val Glu Ser Asn Tyr Val Tyr Ala Asp Lys Ser Ser Tyr Ala Met
290 295 300
Ala Thr Val Thr Gly Gly Ala Tyr Val Ile Thr Pro Glu Leu Lys Leu
305 310 315 320
Ala Val Lys Val Ala Thr Val Asp Tyr Thr Ser Glu Gly Glu Gly Val
325 330 335
Ile Gln Gly Asp Pro Val Ile Glu Tyr Leu Lys Arg His Gly Ser Ala
340 345 350
Leu Asp Gln Pro Ile Met Leu Glu Asn Gly Gly Tyr Ser Leu Thr Lys
355 360 365
Ala Gly Gln Cys Thr Phe Thr Thr Pro Ala Ser Asn Leu Asp Tyr Ser
370 375 380
Leu Thr Ser Gly Asp Thr Asn Pro Ile His Thr Asn Pro Tyr Phe Ala
385 390 395 400
Ser Leu Ala Tyr Leu Pro Gly Thr Ile Thr His Gly Met His Ser Ser
405 410 415
Ala Arg Thr Arg Lys Phe Val Glu Gln Val Ala Ala Asp Asn Val Gly
420 425 430
Ala Arg Val Arg Lys Tyr Glu Val Gly Phe Thr Ala Met Cys Leu Pro
435 440 445
Ser Arg Lys Met Glu Val Arg Leu Lys His Val Gly Met Thr Ala Asp
450 455 460
Gly Asn Arg Leu Ile Lys Val Glu Thr Val Asp Val Glu Gly Gly Asn
465 470 475 480
Val Val Leu Ser Gly Thr Ala Glu Val Ala Gln Ala Pro Thr Ala Tyr
485 490 495
Val Phe Thr Gly Gln Gly Ser Gln Glu Pro Gly Met Gly Met Glu Leu
500 505 510
Tyr Ala Asn Ser Pro Val Ala Arg Ala Val Trp Asp Glu Ala Asp Arg
515 520 525
His Leu Gly Glu Val Tyr Gly Phe Ser Ile Leu Glu Ile Val Arg Thr
530 535 540
Asn Pro Lys Glu Lys Thr Val His Phe Gly Gly Leu Lys Gly Gln Ala
545 550 555 560
Thr Arg Gln Lys Tyr Met Asp Met Ser Tyr Thr Thr Thr Asp His Glu
565 570 575
Gly Asn Val Lys Thr Leu Pro Leu Phe Gly Asp Ile Asp Leu Arg Thr
580 585 590
Ser Arg Tyr Thr Phe Ser Ser Pro Thr Gly Leu Leu Tyr Ala Thr Gln
595 600 605
Phe Ala Gln Ile Ala Leu Val Val Thr Glu Lys Ala Ala Phe Glu Asp
610 615 620
Met Arg Ala Lys Gly Leu Val Gln Lys Asp Cys Val Phe Ala Gly His
625 630 635 640
Ser Leu Gly Glu Tyr Ser Ala Leu Ala Ser Ile Ala Asp Ile Leu Pro
645 650 655
Ile Ser Ala Leu Val Asp Val Val Phe Tyr Arg Gly Ile Thr Met Gln
660 665 670
Arg Ala Val Glu Arg Asp His Leu Asn Arg Ser Ser Tyr Gly Met Val
675 680 685
Ala Val Asn Pro Ser Arg Ile Gly Lys Ser Phe Gly Asp Ala Ala Leu
690 695 700
Arg Glu Val Val Asp Thr Ile Ala Arg Arg Gly Asn Ile Leu Ile Glu
705 710 715 720
Val Val Asn Tyr Asn Val Glu Gly Gln Gln Tyr Val Val Ala Gly His
725 730 735
Leu Val Ala Leu Gln Ser Leu Thr Asn Val Leu Asn Phe Leu Lys Ile
740 745 750
Gln Lys Ile Asp Leu Ala Lys Leu Thr Glu Thr Met Ser Ile Glu Gln
755 760 765
Val Lys Glu His Leu Cys Glu Ile Val Asp Glu Cys Val Gln Lys Ala
770 775 780
Arg Asp Leu Gln Ala Lys Thr Gly Phe Ile Thr Leu Glu Arg Gly Phe
785 790 795 800
Ala Thr Ile Pro Leu Pro Gly Ile Asp Val Pro Phe His Ser Arg Tyr
805 810 815
Leu Trp Ala Gly Val Met Pro Phe Arg Thr Tyr Leu Ser Lys Lys Val
820 825 830
Asn Pro Ala His Phe Asn Ala Asp Leu Leu Val Gly Arg Tyr Ile Pro
835 840 845
Asn Leu Thr Ala Val His Tyr Glu Val Ser Lys Glu Tyr Ala Glu Arg
850 855 860
Ile His Thr Gln Thr Ser Ser Pro Arg Leu Asn Lys Ile Leu Lys Ala
865 870 875 880
Trp Asp Glu Glu Arg Trp Gly Ala Pro Glu Asn Arg Asn Lys Leu Gly
885 890 895
Tyr Ala Ile Leu Ile Glu Leu Leu Ala Tyr Gln Phe Ala Ser Pro Val
900 905 910
Arg Trp Ile Glu Thr Gln Asp Ile Leu Phe Arg Asp Phe Lys Phe Glu
915 920 925
Arg Leu Val Glu Leu Gly Pro Ser Pro Thr Leu Thr Gly Met Ala Thr
930 935 940
Arg Thr Gln Lys Leu Lys Tyr Asp Ala His Asp Ser Ser Val Gly Ile
945 950 955 960
Lys Arg Ser Ile Tyr Cys Ile Ala Lys His Gln Lys Glu Ile Tyr Tyr
965 970 975
Gln Phe Asp Asp Val Ala Gly Glu Glu Ala Pro Ala Pro Ala Ala Val
980 985 990
Ala Pro Ser Ala Pro Ala Pro Lys Ala Ala Pro Val Ala Ala Ala Pro
995 1000 1005
Pro Pro Pro Ala Pro Val Ala Ala Ala Pro Ala Ala Ala Val Ala
1010 1015 1020
Asp Glu Pro Leu Lys Ala Val Asp Thr Leu Arg Ile Ile Ile Ala
1025 1030 1035
Gln Lys Leu Lys Lys Pro Val Gly Glu Val Pro Leu Thr Lys Ser
1040 1045 1050
Ile Lys Glu Leu Val Gly Gly Lys Ser Thr Leu Gln Asn Glu Ile
1055 1060 1065
Leu Gly Asp Leu Gln Gly Glu Phe Ser Ser Ala Pro Glu Lys Gly
1070 1075 1080
Glu Glu Met Pro Leu Gln Glu Leu Gly Ala Ala Leu Gln Gln Gly
1085 1090 1095
Tyr Ser Gly Lys Leu Gly Lys Tyr Thr Thr Gly Val Ile Ser Arg
1100 1105 1110
Met Ile Gly Ala Lys Met Pro Gly Gly Phe Gly Leu Ser Ala Val
1115 1120 1125
Gln Gly His Leu Gly Lys Thr Tyr Gly Leu Gly Ala Gly Arg Ile
1130 1135 1140
Asp Gly Val Leu Leu Phe Ala Val Thr Gln Glu Pro Ala Lys Arg
1145 1150 1155
Leu Ala Asn Glu Gly Glu Ala Lys Ala Trp Val Asp Ser Val Ala
1160 1165 1170
Gln Gly Tyr Ala Ser Met Ala Gly Ile Ser Leu Ala Ala Gly Gly
1175 1180 1185
Gly Ala Ala Ala Ala Ala Pro Ala Met Ala Phe Ala Ala Pro Ala
1190 1195 1200
Ala Ala Gly Gly Gly Ala Pro Ala Lys Thr Ile Leu Ile Leu Gly
1205 1210 1215
Asp Ser Leu Ser Ala Gly Tyr Gly Ile Asn Pro Glu Gln Gly Trp
1220 1225 1230
Val Ala Leu Leu Gln Lys Arg Leu Asp Gln Gln Phe Pro Lys Gln
1235 1240 1245
His Lys Val Ile Asn Ala Ser Val Ser Gly Glu Thr Thr Ser Gly
1250 1255 1260
Ala Leu Ala Arg Leu Pro Lys Leu Leu Thr Thr Tyr Arg Pro Asn
1265 1270 1275
Val Val Val Ile Glu Leu Gly Gly Asn Asp Ala Leu Arg Gly Gln
1280 1285 1290
Pro Pro Gln Met Ile Gln Ser Asn Leu Glu Lys Leu Ile Gln His
1295 1300 1305
Ser Gln Lys Ala Lys Ser Lys Val Val Val Phe Gly Met Lys Ile
1310 1315 1320
Pro Pro Asn Tyr Gly Thr Ala Tyr Ser Gln Ala Phe Glu Asn Asn
1325 1330 1335
Tyr Lys Val Val Ser Gln Thr Tyr Gln Val Lys Leu Leu Pro Phe
1340 1345 1350
Phe Leu Asp Gly Val Ala Gly His Lys Ser Leu Met Gln Asn Asp
1355 1360 1365
Gln Ile His Pro Asn Ala Lys Ala Gln Ser Ile Leu Leu Asn Asn
1370 1375 1380
Ala Tyr Pro Tyr Ile Lys Gly Ala Leu Ala Gly Gly Gly Gly Gly
1385 1390 1395
Gly Ala Ala Val Gly Gly Ala Gly Phe Met Ile Asn Thr Glu Gln
1400 1405 1410
Leu Asp Lys Met Gln Glu Lys Gln Asp Asn Phe Val Ser Gln Gln
1415 1420 1425
Val Glu Leu Phe Leu Arg Tyr Leu Gly Lys Asp Ser Arg Glu Gly
1430 1435 1440
His Arg Leu Ala Asp Met Gln Lys Ala Glu Val Ala Asn Leu Gln
1445 1450 1455
Glu Lys Leu Asp Ser Ile Ala Arg Glu His Gly Asp Ala Tyr Val
1460 1465 1470
Gln Gly Ile Gln Pro Val Phe Asp Pro Leu Lys Ala Arg His Phe
1475 1480 1485
Asn Ser Ser Trp Asn Trp Val Arg Gln Asp Ala Leu Met Met Trp
1490 1495 1500
Met Asp Ile Leu Phe Gly Arg Leu Thr Thr Val Asp Arg Asp Ile
1505 1510 1515
Thr Ala Arg Cys Leu Val Ile Met Asn Arg Ala Asp Pro Ser Leu
1520 1525 1530
Ile Asp Tyr Met Gln Tyr Thr Ile Asp Asn Thr Pro Val Glu Arg
1535 1540 1545
Gly Glu His Tyr Val Leu Ala Lys Gln Phe Gly Gln Gln Leu Leu
1550 1555 1560
Asp Asn Cys Arg Glu Met Ile Gly Gln Ala Pro Leu Tyr Lys Asp
1565 1570 1575
Val Thr Phe Pro Thr Ala Pro Lys Thr Thr Val Asn Ala Lys Gly
1580 1585 1590
Asp Ile Ile Thr Glu Glu Val Asn Arg Pro Gly Val Ser Arg Leu
1595 1600 1605
Glu Lys Tyr Val Ala Glu Met Ala Ala Gly Ser Lys Val Thr Val
1610 1615 1620
Ala Ser Val Asn Leu Asp Lys Val Gln Glu Gln Val Glu Lys Leu
1625 1630 1635
Tyr Lys Leu Val Lys Ser Gln Pro Gln Ile Ser Lys Gln His Met
1640 1645 1650
Thr Ser Ile Lys Ser Leu Tyr Ala Glu Val Val Arg Gly Leu Gly
1655 1660 1665
Lys Asp Ala Gly Pro Pro Pro Val His Lys Ala Gly Thr Arg Ala
1670 1675 1680
Arg Arg Pro Ser Ser Gln Phe Leu Arg Pro Ala Ala Val Ser Glu
1685 1690 1695
Ala Thr Phe Leu Pro Glu Asp Lys Val Pro Leu Leu His Leu Lys
1700 1705 1710
Arg Lys Ile Gly Asn Asp Trp Gln Tyr Ser Ser Lys Leu Thr Ser
1715 1720 1725
Leu Tyr Leu Asp Ile Leu Lys Glu Ile Ala Thr Ser Gly Val Thr
1730 1735 1740
Phe Glu His Lys Asn Ala Leu Met Thr Gly Val Gly Lys Gly Ser
1745 1750 1755
Ile Gly Ile Glu Ile Val Lys Gly Leu Leu Ala Gly Gly Ala Arg
1760 1765 1770
Val Val Ile Thr Thr Ser Arg Tyr Ser Arg Ser Thr Val Glu Tyr
1775 1780 1785
Tyr Gln Ala Ile Tyr Gln Glu Val Gly Ser Lys Gly Ser Ser Leu
1790 1795 1800
Thr Val Val Pro Phe Asn Gln Gly Ser Lys Gln Asp Val Glu Ala
1805 1810 1815
Leu Val Asp Phe Ile Tyr Ser Lys Asp Lys Gly Leu Gly Met Asp
1820 1825 1830
Leu Asp Tyr Ile Leu Pro Phe Ala Ala Leu Pro Glu Asn Gly Arg
1835 1840 1845
Glu Ile Asp Gly Ile Asp Asp Arg Ser Glu Leu Ala His Arg Ile
1850 1855 1860
Met Leu Thr Asn Leu Leu Arg Leu Leu Gly Ala Val Lys Ser Lys
1865 1870 1875
Lys Ala Ala Leu Lys Leu Thr Thr Arg Pro Thr Glu Val Val Leu
1880 1885 1890
Pro Leu Ser Pro Asn His Gly Leu Phe Gly Asn Asp Gly Leu Tyr
1895 1900 1905
Ser Glu Ser Lys Ile Ser Leu Glu Thr Leu Phe Asn Arg Trp Ser
1910 1915 1920
Ser Glu Ser Trp Gly Glu Tyr Leu Cys Leu Ala Gly Ala Val Ile
1925 1930 1935
Gly Trp Thr Arg Gly Thr Gly Leu Met Ser Ala Thr Asn Ser Val
1940 1945 1950
Ala Glu Gly Ile Glu Ala Gln Gly Cys Arg Thr Phe Ser Ala Lys
1955 1960 1965
Glu Met Ala Phe Asn Ile Leu Gly Leu Met His Pro Leu Val Phe
1970 1975 1980
Asp Val Ala Gln Ile Glu Pro Val Trp Ala Asp Leu Asn Gly Gly
1985 1990 1995
Met Asp Lys Leu Pro Asp Leu Ala Asn Leu Thr Thr Glu Ile Arg
2000 2005 2010
Lys Lys Leu Asn Leu Thr Ala Ser Thr Arg Arg Ala Ile Ala Lys
2015 2020 2025
Asp Asn Ser Phe Asp Tyr Lys Val Ala His Gly Pro Ala Met Glu
2030 2035 2040
Gln Ile His Gln Arg Ile Asn Val Ala Pro Arg Ala Asn Phe Ser
2045 2050 2055
Leu Pro Phe Pro Glu Leu Lys Pro Ile Asp Ala Lys Ser Glu Leu
2060 2065 2070
Ala Lys Leu Arg Gly Leu Ile Asp Leu Glu Lys Val Val Val Met
2075 2080 2085
Thr Gly Tyr Ala Glu Val Gly Pro Phe Gly Ser Ser Arg Thr Arg
2090 2095 2100
Trp Glu Met Glu Ala Asn Gly Thr Phe Ser Ile Gln Gly Thr Leu
2105 2110 2115
Glu Leu Ala Tyr Val Met Gly Leu Ile Lys His Phe Glu Gly Arg
2120 2125 2130
Leu Lys Asp Gly Thr Leu Tyr Val Gly Trp Val Asp Ala Lys Thr
2135 2140 2145
Asn Glu Pro Leu Asp Asp Lys Asp Val Lys Ala Ala Tyr Glu Lys
2150 2155 2160
His Ile Leu Ala His Thr Gly Ile Arg Leu Ile Glu Pro Glu Ile
2165 2170 2175
Phe Asn Gly Tyr Asp Pro Lys Arg Lys Gly Phe Thr Gln Glu Ile
2180 2185 2190
Glu Ile Gln His Asp Leu Glu Pro Ile Glu Ala Ser Glu Glu Asp
2195 2200 2205
Ala Ala Arg Phe Lys Arg Glu His Gly Ala Leu Val Asp Val Tyr
2210 2215 2220
Thr Glu Asp Gly Ser Lys Phe Phe Val Lys Phe Lys Lys Gly Ala
2225 2230 2235
Lys Leu His Ile Pro Lys Ala Val Ala Phe Asp Arg Leu Val Ala
2240 2245 2250
Gly Gln Ile Pro Thr Gly Trp Ser His Lys Ala Phe Gly Ile Pro
2255 2260 2265
Asp Asp Ile Ala Ser Gln Val Asp Arg Thr Ser Leu Trp Ala Leu
2270 2275 2280
Val Ser Val Ala Glu Ala Leu Met Met Ala Gly Ile Thr Asp Pro
2285 2290 2295
Tyr Glu Leu Tyr Lys Trp Ile His Pro Ser Glu Val Gly Ser Ser
2300 2305 2310
Leu Gly Ser Gly Met Gly Gly Ile Thr Ser Ile Ser Lys Met Phe
2315 2320 2325
Arg Asp Arg Arg Glu Glu Lys Asp Val Gln Lys Asp Ile Leu Gln
2330 2335 2340
Glu Thr Phe Ile Asn Thr Val Ala Gly Trp Val Asn Leu Leu Leu
2345 2350 2355
Leu Ser Ser Ser Gly Pro Ile Lys Ile Pro Val Gly Ala Cys Ala
2360 2365 2370
Thr Ala Leu Gln Ser Val Glu Ile Ala Cys Asp Thr Ile Leu Ser
2375 2380 2385
Gly Lys Ala Lys Ile Met Val Ser Gly Gly Tyr Asp Asp Phe Ser
2390 2395 2400
Glu Glu Gly Ser Tyr Glu Phe Ala Asn Met Lys Ala Thr Ser Asn
2405 2410 2415
Ser Glu Thr Glu Phe Ala Ala Gly Arg Glu Pro Asn Glu Met Ser
2420 2425 2430
Arg Pro Thr Thr Ser Thr Arg Ala Gly Phe Met Glu Ser Met Gly
2435 2440 2445
Cys Gly Ala Gln Val Leu Met Ser Ala Lys Thr Ala Ile Glu Met
2450 2455 2460
Gly Ala Thr Ile Tyr Gly Ile Val Ala Tyr Thr Ala Thr Ala Thr
2465 2470 2475
Asp Lys Ala Gly Arg Ser Ile Pro Ala Pro Gly Arg Gly Val Met
2480 2485 2490
Gly Thr Ala Arg Glu Ile Thr Ser Lys Tyr Pro Ser Pro Ile Leu
2495 2500 2505
Asp Val Thr Tyr Arg Arg Arg Gln Leu Glu Phe Arg Arg Lys Gln
2510 2515 2520
Ile Ser Gln Trp Leu Glu Asn Glu Thr Glu Leu Leu Lys Phe Glu
2525 2530 2535
Val Ser Ser His Gly Gln Ala Thr Lys Leu Pro Asp Asp Tyr Val
2540 2545 2550
Ser Glu Arg Leu Ala Ser Ile Glu Arg Glu Ala Lys Arg Gln Glu
2555 2560 2565
Ala Glu Ala Leu Ala Thr Tyr Gly Met Leu Ala Gly Gln Asp Pro
2570 2575 2580
Thr Ile Ala Pro Leu Arg Arg Ala Leu Ala Val Trp Gly Leu Thr
2585 2590 2595
Ile Asp Asp Val Gly Val Ala Ser Phe His Gly Thr Ser Thr Val
2600 2605 2610
Ala Asn Asp Lys Asn Glu Ser Asn Ala Tyr Asn Glu Gln Phe Arg
2615 2620 2625
His Leu Gly Arg Ala Lys Gly Asn Ala Cys Pro Val Ile Ala Gln
2630 2635 2640
Lys Trp Leu Thr Gly His Pro Lys Gly Gly Ala Ala Ala Trp Met
2645 2650 2655
Leu Asn Gly Leu Ala Gln Val Ile Gln Ser Gly Leu Val Pro Gly
2660 2665 2670
Asn Arg Asn Ala Asp Asn Ile Gly Glu Glu Leu Arg Ala Phe Glu
2675 2680 2685
Tyr Leu Leu Tyr Pro Ser Lys Ser Ile Gln Thr Asp Gly Ile Lys
2690 2695 2700
Ala Gly Leu Leu Thr Ser Phe Gly Phe Gly Gln Val Gly Gly Gln
2705 2710 2715
Ala Leu Ile Val His Pro Ser Leu Leu Ile Gly Ala Leu Glu Pro
2720 2725 2730
Ala Gln Phe Glu Ala Tyr Lys Lys Leu Asn Asp Gln Arg Lys Lys
2735 2740 2745
Trp Ser Tyr Arg Arg Phe Asn Asp Phe Phe Thr Asn Gly Lys Leu
2750 2755 2760
Val Ile Ile Lys Asp Gly Thr Pro Phe Thr Pro Glu Gln Glu Asn
2765 2770 2775
Thr Thr Leu Leu Asn Pro Leu Val Arg Ala Val Pro Asp Lys Thr
2780 2785 2790
Gly Ser Tyr Ser Met Pro Lys Glu Phe Pro Ala Thr Val Pro Arg
2795 2800 2805
Ser Asn Asn Ala Glu Val Ala Asn Lys Leu Val Ser Ala Ala Val
2810 2815 2820
Gly Gly Ala Phe Gly Val Gly Thr Asp Val Glu Leu Ile Ser Ala
2825 2830 2835
Val Pro Thr Ser Glu Ser Phe Leu Glu Arg Asn Phe Thr Gln Asp
2840 2845 2850
Glu Ile Ala Tyr Cys Lys Ala Ala Pro Asp Phe Arg Ala Ser Leu
2855 2860 2865
Ala Ala Arg Trp Ser Ala Lys Glu Ala Thr Phe Lys Ala Leu Lys
2870 2875 2880
Thr Glu Ser Lys Gly Ala Ala Ala Ser Met Gln Asp Ile Glu Val
2885 2890 2895
Val Ser Thr Ser Gln Gly Pro Thr Ile Lys Leu His Gly Glu Val
2900 2905 2910
Glu Lys Ile Ala Gln Ala Ala Gly Ile Thr Ala Phe Glu Val Ser
2915 2920 2925
Leu Ser His Ser Glu Asp Val Ala Cys Ala Val Val Ile Ala Gln
2930 2935 2940
Lys
<210> 12
<211> 4158
<212> PRT
<213> Aplanochytrium kerguelense
<400> 12
Met Glu Gln His Lys Thr Glu Lys Leu Ser Ala Ala Asp Glu Lys Leu
1 5 10 15
Ser Glu Ser Thr Ile His Trp Leu Ala Ala Thr Asp Val Pro Ser Asn
20 25 30
Phe Val Leu Phe Ser Gly Gln Gly Tyr Gln Tyr Phe Asp Glu Leu Arg
35 40 45
Glu Leu Tyr Glu Thr Gly Thr Asp Glu Val Lys Gly Phe Leu Leu Leu
50 55 60
Ala Ser Lys Thr Leu Gln Glu Glu Ile Cys Ser Glu Lys Ala Lys Thr
65 70 75 80
Val Ala Phe Lys Phe Val Asn Glu Leu Asp Phe Thr Lys Trp Ile Glu
85 90 95
Glu Asp Ile Asp Val Asp Gln Thr Tyr Ile Phe Ser Ala Pro Val Ser
100 105 110
Phe Pro Leu Ile Phe Val Ala Gln Val Ala Arg Tyr Leu Gln Thr Leu
115 120 125
Asn Leu Leu Arg Thr Asp His Lys Gln Phe Ile Gln Thr Ile Lys Gly
130 135 140
Ser Ser Gly His Ser Gln Gly Ile Val Ala Ala Val Leu Ile Ser Thr
145 150 155 160
Ser Pro Asp Asn Asp Arg Ile Val Glu Asn Ala Val Lys Leu Cys Arg
165 170 175
Tyr Met Leu Trp Gln Gly Leu Arg Cys His Glu Ser Ser Val Asn Arg
180 185 190
Phe Ser Gly Lys Asn His Lys Ser Lys Lys Ser Lys Val Tyr Ser Phe
195 200 205
Asp Glu Ser Thr Arg Met Val Lys Tyr Ser Pro Met Leu Ala Val Asn
210 215 220
Gly Leu Thr Glu Ser Leu Leu Asn Asp Phe Met Lys Lys Thr Glu Ser
225 230 235 240
Ile Ile Glu Ala Arg Arg Thr Ile Gln Gln Lys Tyr Asn Arg Arg Glu
245 250 255
Ser Phe Phe Glu Thr Pro Thr Glu Lys Asp Glu Lys Ser Lys Arg Leu
260 265 270
Asn Met Phe Gln Ile Ala Leu Ala Asn Gly Pro Lys Ser Phe Val Val
275 280 285
Ser Gly Ala Pro Lys Thr Leu Ile Glu Leu Glu Gln Arg Ile Lys Glu
290 295 300
Ser Thr Thr Ser Ser Pro Asn Ser Gln Thr Arg Ile Pro Phe Ser Lys
305 310 315 320
Arg Lys Pro Val Val Lys Met Tyr Phe Leu Lys Val Gly Ala Ala Phe
325 330 335
His Thr Glu Ile Cys Asn Asp Ala Ser Ser Lys Leu Lys Ala Asp Val
340 345 350
Val Gln Met Lys Leu Ser Ile Ser Pro Lys Glu Ile Leu Ile Pro Val
355 360 365
Tyr His Thr Lys Asp Gly Ser Asn Leu Ser Glu Ile Ser Ala Asp Val
370 375 380
Asn Val Val Asp Leu Leu Ile Asp Met Gln Thr Ser Gln Val Asn Asp
385 390 395 400
Phe Arg Lys Thr Leu Lys Ser Ile Ser Ser Gln Asn Gly Val Ser Asn
405 410 415
Val Ile Asp Phe Gly Pro Gly Asp Gly Thr Ala Lys Leu Cys Leu Lys
420 425 430
Arg Lys Ser Gly Thr Gly Ile Leu Val Val Ala Ala Thr Gly Pro Leu
435 440 445
Arg Asn Arg Arg Cys Ala Tyr Gly Arg Asn Leu Val Gly Leu Asn Ser
450 455 460
Val Leu Leu Glu Lys Asn Pro Ile Leu Gly Met Asn Trp Gly Glu Glu
465 470 475 480
Phe Lys Pro Arg Leu Ser Ser Arg Lys Asn Asp Asn Gln Ile Ile Val
485 490 495
Gly Thr Arg Phe Ser Asp Leu Thr Gly Lys Pro Pro Val Ile Leu Pro
500 505 510
Gly Met Thr Pro Thr Thr Ser Phe His Gly Ile Asp Leu Val Ala Ala
515 520 525
Cys Ser Asn Gly Gly Tyr His Gly Glu Leu Ala Gly Gly Gly Leu Pro
530 535 540
Leu Pro Glu Tyr Phe Lys Ala Lys Ile Asp Glu Leu Val Ser Lys Gln
545 550 555 560
Asn Ser Gly Val Gly Ile Asn Ile Asn Met Leu Tyr Leu Asn Ser Tyr
565 570 575
Leu Trp Gly Phe Gln Phe Ser Thr Ala Ile Gln Met Ala Lys Asp Gly
580 585 590
Tyr Pro Ile Glu Ser Ile Thr Cys Ala Ala Gly Val Pro Thr Thr Glu
595 600 605
Lys Ala Lys Glu Ile Met His Gln Leu Lys Glu Ala Gly Ile Lys Tyr
610 615 620
Ile Ala Phe Lys Pro Gly Ser Arg Ala Ala Ile Met Asp Val Leu Ser
625 630 635 640
Ile Ala Arg Glu Asn Pro Asn Ser Ser Ile Val Leu Gln Trp Thr Gly
645 650 655
Gly Arg Gly Gly Gly His His Ser Phe Glu Asp Phe His Ala Pro Leu
660 665 670
Leu Asp Thr Tyr Gln Glu Ile Arg Glu His Pro Asn Val Val Leu Val
675 680 685
Val Gly Ser Gly Phe Gly Asp Ala Glu Lys Ser Tyr Pro Tyr Leu Asp
690 695 700
Gly Ser Trp Ser Thr Leu Pro Pro His Asn Cys Glu Ser Arg Met Pro
705 710 715 720
Cys Asp Ala Val Leu Phe Gly Ser Arg Cys Met Val Ala Lys Glu Ala
725 730 735
Ala Thr Ser Pro Glu Val Lys Gln Leu Ile Val Asn Ala Lys Gly Val
740 745 750
Arg Glu Asn Glu Ser Trp Glu Met Ser Tyr Glu Asn Asp Ala Gly Gly
755 760 765
Ile Leu Thr Val Thr Ser Glu Leu Gly Glu Pro Ile His Lys Ile Asn
770 775 780
Asn Arg Gly Met Lys Leu Trp Arg Glu Phe Asp Arg Lys Tyr Phe Ser
785 790 795 800
Leu Pro Ser Gly Gln Glu Arg Glu Lys Ala Ile Val Lys Asp Lys Lys
805 810 815
Glu Ile Ile Arg Arg Ile Asn Ala Asp Phe Gln Lys Val Tyr Phe Gly
820 825 830
Arg Lys Ala Asp Gly Thr Val Val Asp Ile Ala Tyr Met Thr Tyr Gly
835 840 845
Glu Ile Leu Arg Arg Met Val Asp Leu Met Tyr Val Thr Gly Gly Gly
850 855 860
Asp Gly Lys Gln His Arg Phe Ala Pro Asn Arg Trp Ile Asp Val Thr
865 870 875 880
Tyr Gln Thr Arg Thr Phe Lys Phe Leu Val Arg Thr Glu Arg Arg Phe
885 890 895
Leu Arg Asp Lys Lys Met Ala Phe Ala Thr Asp Met Arg Lys Leu Glu
900 905 910
Ser Phe Pro Ile Lys Cys Ile Asp Glu Phe Val Lys Cys Tyr Pro Gln
915 920 925
Val Asp Lys Val Leu Val Ser Asp Glu Asp Ala Gly Tyr Phe Ile Glu
930 935 940
Leu Cys Arg Asn Leu Arg Asn Gly Lys Pro Val Asn Phe Ile Pro Gln
945 950 955 960
Ile Asp Gly Ser Leu Ala Tyr Trp Phe Lys Lys Asp Ser Leu Trp Cys
965 970 975
Ser Glu Gln Ile Asp Ala Val Pro Asp Asn Asp Pro Gly Arg Val Cys
980 985 990
Ile Leu His Gly Pro Val Ala Ala Gln Tyr Ser Val Val Ser Asp Glu
995 1000 1005
Pro Ile Cys Glu Ile Leu Gly Asn Ile His Lys Gly Tyr Val Asp
1010 1015 1020
Lys Leu Lys Glu Glu Lys Tyr Glu Val Ala Lys Val Glu Thr Ile
1025 1030 1035
Gly Lys Leu Pro Glu Tyr Lys Gly Ser Asn Lys Ser Phe Arg Lys
1040 1045 1050
Phe Leu Arg Tyr Gly Trp Phe Asn Ala Leu Phe Gln Ser Thr Val
1055 1060 1065
Met Val Lys Glu Lys Lys Trp Val Arg Asn Ile Ile Pro Gly Ile
1070 1075 1080
Ile Gln Thr Glu Asn Glu Asn Ile Val Phe Ser Glu Arg Thr Ala
1085 1090 1095
Met Asn Asp Met Cys Leu Ile Glu Val Leu Leu Asn Asp Ser Gln
1100 1105 1110
Arg Thr Leu Ala Phe Ile Glu Phe Glu Gly Lys His Ile Ser Val
1115 1120 1125
Ser Leu Gln Asp Ser Glu Pro Val Asn Ser Thr Leu Thr Leu Tyr
1130 1135 1140
Phe Glu Tyr His Pro Glu Thr Pro His Ser Pro Leu Phe Gln Val
1145 1150 1155
Thr Asn Gly Ser Lys Ser Ile Ala Arg Ser Ile Lys Gln Tyr Tyr
1160 1165 1170
Arg Asn Ile Trp Asp Cys Gln Asp Val Val Ser Ile His Asp Thr
1175 1180 1185
Phe Ser Glu Ser Phe Thr Val Thr Arg Glu Asp Ile Glu Arg Phe
1190 1195 1200
Asn Gly Ala Ile Gln Tyr Ala Val Asn Thr Ser Glu Gly Thr Leu
1205 1210 1215
Asp Phe Ala Ile Val Ala Gly Trp Lys Ser Leu Ile Lys Ala Leu
1220 1225 1230
Phe Ser Lys Glu Ile Asp Gly Ser Leu Leu Asp Leu Val His Ile
1235 1240 1245
Ser His Ser Tyr Lys Leu Leu Val Glu Glu Lys Glu Arg Thr Leu
1250 1255 1260
Val Gln Ala Gly Glu Lys Ile Cys Ser Glu Ser Arg Val Ile Ser
1265 1270 1275
Val Gln Ile Leu Pro Gly Val Gly Lys Ser Ile Thr Val Lys Gly
1280 1285 1290
Val Leu Ser Arg Asn Thr Val Lys Trp Val Glu Val Gln Ser Glu
1295 1300 1305
Phe Leu Ile Arg Gly Asn Phe Phe Asp Phe Gln Lys Thr Phe Lys
1310 1315 1320
Glu Tyr Ser Tyr Lys Ala Glu Val Leu Cys Lys Asp Asp Thr Val
1325 1330 1335
Pro Gln Ile Leu Asn Ser Gln Glu Trp Leu Phe Leu Asp Ser Gly
1340 1345 1350
Val Arg Pro Leu Ser Lys His Asp Lys Val Val Phe Gln Ile Asp
1355 1360 1365
Arg Val Val Glu Lys Arg Lys Ser Val Thr Asn Ile Ser Asp Ile
1370 1375 1380
Leu Val Ala Gly Asn Val Val Arg Ile Asn Thr Pro Glu Glu Ile
1385 1390 1395
Asp Gly Phe Ile Glu Leu Lys Asp Gln Gly Ser Pro Lys Gly Ile
1400 1405 1410
Ile Ile Gly Lys Val Asp Leu Ser Leu Glu Asn Ala Asn Leu Ser
1415 1420 1425
Glu Asn Pro Ile Ile Thr Tyr Leu Asp Ser Ile Lys Glu Ser Lys
1430 1435 1440
Leu His Gly Ser Val Phe Glu Ser Gly Gly Tyr Thr Val Met Pro
1445 1450 1455
Arg Pro Glu Leu Val Thr Ala Pro Ser Ile Val Glu Ser Ile Val
1460 1465 1470
Tyr Ala Ser Ala Ser Arg Asp Met Asn Pro Ile His Arg Asn Arg
1475 1480 1485
Thr Phe Ala Lys Leu Ala Gly Leu Pro Gly Gly Ser Thr Ile Val
1490 1495 1500
His Gly Met Trp Thr Ala Ala Met Ser Arg Arg Ile Leu Glu Leu
1505 1510 1515
His Thr Val Leu Gly Asp His Arg Arg Ile Ser Thr Tyr Ser Val
1520 1525 1530
Lys Phe Thr Gly Met Val Phe Pro Glu Asp Lys Leu Ala Val Met
1535 1540 1545
Val Lys His Val Gly Val Thr Glu Gly Arg Leu Ile Leu Asp Val
1550 1555 1560
Asp Val Ser Lys Val Glu Thr Gln Glu Arg Val Leu His Gly Arg
1565 1570 1575
Ala Glu Val Glu Gly Pro Ser Thr Ser Phe Leu Phe Thr Gly Gln
1580 1585 1590
Gly Ser Ala Lys Val Gly Met Gly Met Asp Ser Tyr Glu Glu Lys
1595 1600 1605
Gln Ser Ala Arg Ala Val Trp Gln Thr Ala Asp Lys Phe Leu Arg
1610 1615 1620
Lys Lys Phe Gly Phe Ser Ile Val Asp Ile Val Lys Asn Asn Pro
1625 1630 1635
Lys Glu Leu Thr Val Arg Phe Gly Gly Ser Gln Gly Lys Val Ile
1640 1645 1650
Arg Glu Asn Tyr Arg Ala Ile Lys Arg Glu Asp Gly Ser Gln Leu
1655 1660 1665
Ile Arg Glu Ile His Ser Asn Thr Thr Ser Phe Thr Phe Arg Ser
1670 1675 1680
Pro Asn Gly Leu Leu Phe Ala Thr Gln Phe Ser Gln Pro Ala Leu
1685 1690 1695
Val Leu Val Gln Lys Ala Ala Phe Glu Glu Met Leu Gln Gly Gly
1700 1705 1710
Phe Val Pro Ser Asn Ser Met Phe Ala Gly His Ser Leu Gly Glu
1715 1720 1725
Tyr Ala Ala Leu Ala Ser Tyr Ala Asn Val Leu Ser Ile Glu Asp
1730 1735 1740
Leu Val Glu Thr Val Phe Leu Arg Gly Met Val Met Gln Asn Ala
1745 1750 1755
Val Ala Arg Asp Asp Glu Gly Thr Ser Asp Tyr Gly Met Val Ala
1760 1765 1770
Ala Asn Pro Ser Arg Val Ser Lys Thr Phe Ser Ala Gln Leu Leu
1775 1780 1785
Met Glu Ile Val Gln Lys Ile Asp Asp Glu Ser Asp Asp Ser His
1790 1795 1800
Leu Leu Gln Val Val Asn Phe Asn Ile Arg Asp Ala Gln Tyr Val
1805 1810 1815
Ile Ala Gly Asn Leu Glu Leu Leu Asp Ser Leu Ser Asn Val Leu
1820 1825 1830
Asn Val Ile Ser Asn Ser His Asp Lys Thr Phe Asp Arg Ser Ile
1835 1840 1845
Ile Gln Ala Ala Met Ile Lys Ala Glu Asn Arg Arg Arg Ser Cys
1850 1855 1860
Lys Glu Asn Asn Gln Pro Phe Arg Leu Lys Arg Gly Ile Ala Thr
1865 1870 1875
Ile Pro Leu His Gly Ile Asp Val Pro Phe His Ser Arg Gln Leu
1880 1885 1890
Leu Asn Gly Val Pro Ala Phe Arg Ser Leu Leu Glu Pro Arg Phe
1895 1900 1905
Thr Glu Glu Met Ile Gln Lys Tyr Leu Pro Arg Leu Ile Gly Lys
1910 1915 1920
Tyr Ile Pro Asn Val His Ala Glu Pro Phe Ser Ile Ser Lys Glu
1925 1930 1935
Tyr Ile Glu Lys Val Ala Ser Val Thr Lys Ser Pro Ala Leu Tyr
1940 1945 1950
Lys Leu Leu Ser Thr Tyr Ser Thr Met Cys Asp Ser Ser Lys Ala
1955 1960 1965
Arg Ile Leu Leu Ile Glu Leu Leu Ala His Gln Phe Ala Met Pro
1970 1975 1980
Val Gln Trp Ile Asp Thr Gln Asp Tyr Ile Leu Ser Arg His Thr
1985 1990 1995
Gln Arg Ile Ile Glu Met Gly Pro Ala Pro Thr Leu Val Gly Met
2000 2005 2010
Ala Lys Lys Thr Leu Gln Ser Arg Ile Tyr Gly Asp Glu Glu Asp
2015 2020 2025
Tyr Lys Pro Ser Ile Leu Trp Trp Asn Gln Asn Gln Glu Glu Ile
2030 2035 2040
Phe Tyr Asn Leu Asp Asn Gln Gly Ile Ser Ile Gln Gln Phe Leu
2045 2050 2055
Gln Glu Leu Lys Gly Thr Glu Asp Asp Glu Ser Gln Ser Asp Asp
2060 2065 2070
Met Ser Ser Tyr Thr Gln Ser Asp Glu Thr Ile Gln Arg Thr Ala
2075 2080 2085
Lys Asn Arg Val Met Pro Glu Ala Asn Ala Asn Asp Lys Glu Lys
2090 2095 2100
Val Gly Asn Glu Leu Gln Lys Lys Glu Val Thr Asn Arg Pro Leu
2105 2110 2115
His Val Leu Arg Val Leu Ile Ser Thr Lys Leu Lys Lys Lys Met
2120 2125 2130
Leu Glu Val Lys Ala Thr Asp Thr Ile Lys Gly Val Ser Gly Gly
2135 2140 2145
Lys Ser Ala Ile Gln Asn Glu Leu Ile Gly Glu Ile Asn Ala Glu
2150 2155 2160
Tyr Ser Ser Phe Arg Asp Ile Glu Asp Ile Ser Glu Val Ser Leu
2165 2170 2175
Ala Glu Leu Ala Arg Arg Ala Pro Ser Arg Gln Glu Thr Val Leu
2180 2185 2190
Gly Lys Val Thr Thr Lys Met Val Asn Lys Met Phe Ser Ser Lys
2195 2200 2205
Leu Pro Gly Lys Phe Gly Pro Asn Asp Thr Lys Glu Tyr Leu Gln
2210 2215 2220
Ser Lys Leu Ser Asp Ala Ser Thr Val Asp Ser Val Ser Leu His
2225 2230 2235
Ala Leu Thr Leu Gln Pro Val Ser Arg Leu Ala Thr Pro Gly Glu
2240 2245 2250
Ala Gln Ala Trp Leu Asp Ser Val Cys Arg Asp Phe Glu Asn Asp
2255 2260 2265
Gln Arg Ile Ser Ile Leu Lys Asn Lys Ser Pro Asn Ser Asn Glu
2270 2275 2280
Ser Val Met Ile Ser Glu Gln Arg Thr Val Gln Asn Asn Phe Val
2285 2290 2295
Gln Ile Lys Thr Ile Leu Ile Leu Gly Asp Ser Leu Ser Ala Gly
2300 2305 2310
Tyr Gly Ile Asn Pro Glu Gln Gly Trp Val Ala Leu Leu Gln Lys
2315 2320 2325
Arg Leu Asp Gln Gln Phe Pro Lys Gln His Lys Val Ile Asn Ala
2330 2335 2340
Ser Val Ser Gly Glu Thr Thr Ser Gly Ala Leu Ala Arg Leu Pro
2345 2350 2355
Lys Leu Leu Thr Thr Tyr Arg Pro Asn Val Val Val Ile Glu Leu
2360 2365 2370
Gly Gly Asn Asp Ala Leu Arg Gly Gln Pro Pro Gln Met Ile Gln
2375 2380 2385
Ser Asn Leu Glu Lys Leu Ile Gln His Ser Gln Lys Ala Lys Ser
2390 2395 2400
Lys Val Val Val Phe Gly Met Lys Ile Pro Pro Asn Tyr Gly Thr
2405 2410 2415
Ala Tyr Ser Gln Ala Phe Glu Asn Asn Tyr Lys Val Val Ser Gln
2420 2425 2430
Thr Tyr Gln Val Lys Leu Leu Pro Phe Phe Leu Asp Gly Val Ala
2435 2440 2445
Gly His Lys Ser Leu Met Gln Asn Asp Gln Ile His Pro Asn Ala
2450 2455 2460
Lys Ala Gln Ser Ile Leu Leu Asn Asn Ala Tyr Pro Tyr Ile Lys
2465 2470 2475
Gly Ala Leu Arg Thr Leu Ser Ser Gly Asp Ser Ser Pro Ala His
2480 2485 2490
Ser Thr Lys Asp Ile Leu Ser Ser Thr Ala Met Lys Lys Leu Lys
2495 2500 2505
Lys Leu Tyr Leu Asp Gln Val Asp Val Phe Asn Thr Phe Leu Gly
2510 2515 2520
Glu Asp Arg Arg Ala Lys Leu Gln Lys Val Leu Thr Glu Glu Thr
2525 2530 2535
Ser Ala Lys Asp Arg Ile Glu Ala Gln Leu Leu Glu Leu Lys Glu
2540 2545 2550
Glu Leu Gly Asp Pro Phe Cys Asp Gly Ile Gln Pro Gln Phe Asn
2555 2560 2565
Glu Ala Arg Ile Arg Ile Tyr Asp Ser Tyr Trp Asn Trp Val Val
2570 2575 2580
Gln Asp Ala Leu Glu Leu His Tyr His Thr Phe Ser Cys Val Leu
2585 2590 2595
Asn Ser Lys Gly Lys Asn Val Asn Ile Pro Asn Ser Ser Asn Ser
2600 2605 2610
Tyr Phe Arg Ala Met Ser Asp Trp Ile Thr Ser Ser Asn Glu Lys
2615 2620 2625
Leu Glu Glu Asn Lys Pro Pro Gln Ala Trp Phe Arg Asn Tyr Leu
2630 2635 2640
Cys Asn Arg Ala Thr Pro Glu Leu Leu Thr Val Val Gln Tyr Phe
2645 2650 2655
Ala Ala Arg Met Asn Glu Gln Gly His Ser Glu Tyr Ala Gln Ala
2660 2665 2670
Val Ser Leu Leu Ala Glu Gln Val Ser Gln Trp Ile Ser Arg Pro
2675 2680 2685
Pro Val His Ile Ala Leu Phe Ala Ser Leu Glu Pro Arg Val Thr
2690 2695 2700
Val Asp Ser Ala Asn Asn Phe Gly Leu Lys Tyr Glu Glu Lys Pro
2705 2710 2715
Arg Lys Phe Val Ala Val Gly Glu Asn Lys Ala Phe Ser Cys Asp
2720 2725 2730
Asn Ala Ser Leu Tyr Val Lys Glu Met Ser Lys Gly Leu Phe Tyr
2735 2740 2745
Asp His Arg Val Ala Ser Lys Val Glu His Pro Ser Gln Ser Val
2750 2755 2760
Tyr Leu Ser Pro Asp Asn Ser Phe Phe Pro Asn Glu Glu Glu Ser
2765 2770 2775
Asp Gln Ile Ser Ser Gly Met Arg Leu Pro Lys Thr Gln Gly Glu
2780 2785 2790
Ile Gln Arg Asp Phe Ser Arg Leu Pro Ala Gly Gln Lys Leu Glu
2795 2800 2805
Val Met Arg Lys Ser Val Asn Arg Ser Ser Asp Thr Asp Glu Glu
2810 2815 2820
Ile Ala Asn Ile Val Arg Gln Asn Tyr Asn Ser Ile His Val Ala
2825 2830 2835
Lys Asn Val Pro Leu Val His Leu Lys Ser Pro Ser Lys Phe Asp
2840 2845 2850
Lys Thr Val Arg Val Leu Asp Glu Pro Leu Thr Ser Met Tyr Leu
2855 2860 2865
Ser Cys Leu His Asp Ile Ala Thr Ser Gly Val Ser Phe Ala Gly
2870 2875 2880
Gln Asn Ala Leu Val Thr Gly Ala Gly Phe Gly Ser Ile Gly Ile
2885 2890 2895
Glu Leu Ile Lys Pro Leu Leu Glu Gly Gly Ala Thr Val Leu Val
2900 2905 2910
Thr Val Arg Leu Asn Arg Thr Asp Glu Gln Met Gln Ile Val Asn
2915 2920 2925
Glu Arg Phe Gln Arg Leu Tyr Glu Glu Phe Gly Ser Arg Gly Ser
2930 2935 2940
Lys Leu Val Leu Val Pro Cys Asn Cys Ala Ser Asn Gln Asp Val
2945 2950 2955
His Ser Leu Ile Arg His Ile Tyr Glu Lys Leu Lys Leu Asp Leu
2960 2965 2970
Asp Phe Ile Phe Pro Phe Ala Ala Ile Gly Glu Gln Gly Lys Asp
2975 2980 2985
Val Ser Glu Ile Gly Ser Lys Ser Glu Val Ala His Arg Leu Met
2990 2995 3000
Leu Thr Asn Thr Ile Arg Leu Leu Gly Ala Val Lys Lys Ala Lys
3005 3010 3015
Glu Asp Arg Cys Ile Glu Thr Arg Pro Ala Leu Val Leu Leu Pro
3020 3025 3030
Cys Ser Pro Asn His Gly Asp Phe Gly Leu Asp Gly Leu Tyr Ala
3035 3040 3045
Glu Ser Lys Leu Gly Leu Glu Ser Leu Val Asn Lys Trp Lys Ser
3050 3055 3060
Glu Glu Trp Gly Asn Tyr Leu Ser Ile Cys Ala Ala Val Ile Gly
3065 3070 3075
Trp Thr Arg Ser Lys Leu Met Trp Gln Asn Asn Val Val Ala Glu
3080 3085 3090
Gly Ile Glu Lys Leu Gly Val Arg Thr Phe Ser Thr Thr Glu Thr
3095 3100 3105
Ala Phe Asn Leu Ile Gly Leu Leu His Pro Asp Ile Val Ser His
3110 3115 3120
Ala Ala Glu Glu Pro Leu Trp Ala Asp Leu Thr Gly Asn Trp Gly
3125 3130 3135
Ala Ala Pro Asp Leu Lys Glu His Ser Lys Arg Ile Arg Thr Ser
3140 3145 3150
Leu Leu Ala Gln Ser Lys Ala Ala Lys Ala Ile Ser Leu Ser Ser
3155 3160 3165
Lys Ser Leu Val Ile Glu Pro Ser Glu Ser Lys Thr Lys Ala Gln
3170 3175 3180
Thr Glu Ile Val Asn Thr Tyr Leu Ala Gly Val Asn Asp Lys Leu
3185 3190 3195
Arg Leu Pro Leu Ala Asn Pro Glu Lys Phe Cys Asn Pro Phe Pro
3200 3205 3210
Lys Ile Pro Ser Gln Glu Arg Met Asp Ser Leu Ala Tyr Leu Lys
3215 3220 3225
His Ser Val Asp Leu Lys Lys Val Val Val Val Val Gly Tyr Gly
3230 3235 3240
Glu Ile Gly Pro Trp Gly Asn Ser Arg Thr Arg Trp Glu Met Glu
3245 3250 3255
Ser Phe Gly Glu Phe Ser Leu Glu Gly Ala Ile Glu Leu Ala Trp
3260 3265 3270
Leu Val Gly Leu Ile Lys Pro Val Thr Gly Pro Leu Lys Asn Asp
3275 3280 3285
Pro Arg Thr Gln Tyr Phe Gly Trp Val Asp Ala Glu Ser Glu Glu
3290 3295 3300
Pro Val Ala Asp His Glu Ile Lys Thr Arg Tyr Glu Lys Val Leu
3305 3310 3315
Leu Gln His Ser Gly Ile Arg Leu Ile Glu Pro Glu Leu Phe Glu
3320 3325 3330
Gly Tyr Asn Pro Lys Lys Lys Ser Ile Leu Arg Gln Val Ala Ile
3335 3340 3345
Ala Glu Asp Met Lys Pro Ile Glu Val Ala Ser Leu Glu Glu Ala
3350 3355 3360
Gln Gln Tyr Val Asn Glu Leu Gly Lys Glu Phe Ile Asp Val Phe
3365 3370 3375
Asn Glu Asn Val Glu Ser Asn Asp Gly Gln Trp Tyr Ile Arg Leu
3380 3385 3390
Lys Ala Gly Ala Val Val Ser Ile Pro Gly Ala Leu Ser Phe Asn
3395 3400 3405
Arg Phe Ile Ala Gly Gln Leu Pro Thr Gly Trp Asp Ala Lys Arg
3410 3415 3420
Leu Gly Ile Pro Asp Asp Ile Ala Asp Ser Val Asp Pro Val Thr
3425 3430 3435
Leu Tyr Ala Leu Val Ser Thr Val Glu Ala Leu Val Cys Ala Gly
3440 3445 3450
Leu Thr Asp Pro Tyr Glu Leu Tyr Gln Tyr Val His Val Ser Gln
3455 3460 3465
Val Gly Asn Thr Ser Gly Gly Gly Met Gly Gly Met Arg Ser Leu
3470 3475 3480
Lys Arg Met Phe Leu Glu Arg Lys Leu Asp Ala Glu Ile Pro Ser
3485 3490 3495
Asp Thr Leu Ala Glu Ser Phe Ile Asn Thr Met Pro Ala Trp Val
3500 3505 3510
Asn Met Leu Leu Leu Ser Ser Ser Gly Pro Ile Lys Thr Pro Val
3515 3520 3525
Gly Ala Cys Ala Thr Ala Ala Glu Ser Val Asp Ile Gly Ile Glu
3530 3535 3540
Thr Ile Leu Ala Gly Lys Ala Arg Val Val Ile Ala Gly Gly Tyr
3545 3550 3555
Asp Asp Phe Cys Glu Thr Gly Ser Asn Glu Phe Ala Met Met Gly
3560 3565 3570
Ala Thr Ser Asn Ser Gln Thr Glu Ala Glu Lys Gly Arg Phe Pro
3575 3580 3585
Arg Glu Ala Ser Arg Pro Met Thr Asp Thr Arg Ala Gly Phe Met
3590 3595 3600
Glu Ser Gln Gly Ala Gly Met Gln Val Leu Met Asp Ala Glu Leu
3605 3610 3615
Ala Ile Gln Met Gly Leu Pro Val Tyr Gly Ile Leu Ala Leu Ser
3620 3625 3630
Asn Thr Ala Thr Asp Arg Gln Gly Arg Ser Val Pro Ala Pro Gly
3635 3640 3645
Arg Gly Ile Leu Thr Thr Ala Arg Glu Val Arg Ser Lys Asn Ser
3650 3655 3660
Lys Ser Ser His Asn Gly Lys Pro Val Glu Asn Pro Leu Leu Ser
3665 3670 3675
Val Lys Phe Arg Lys Lys His Leu Arg Gln Glu Leu Asp Ala Ile
3680 3685 3690
Asp Leu Trp Ala Ser Gln Glu Ile Ser Asn Met Ser Glu Glu Phe
3695 3700 3705
Ser Gln Asn Ser Arg Lys Met Glu Asn Arg Lys Gly Phe Val Glu
3710 3715 3720
Thr Met Arg Lys Lys Lys Arg Ser Ala Ala Phe Glu Thr Trp Gly
3725 3730 3735
Gln Gly Phe Tyr Arg Asn Asp Asp Ser Ile Ala Pro Leu Arg Gly
3740 3745 3750
Ala Leu Ser Val Trp Gly Leu Thr Val Asp Asp Leu Leu Val Gly
3755 3760 3765
Ser Phe His Gly Thr Gly Thr Asn Leu Asn Asp Thr Asn Glu Ser
3770 3775 3780
Ser Leu Val Asn Lys Gln Leu Lys His Leu Asp Arg Lys Glu Gly
3785 3790 3795
Asn Ile Leu Leu Val Val Thr Gln Lys Tyr Leu Thr Gly His Pro
3800 3805 3810
Lys Gly Ala Ala Ala Ala Trp Met Leu Asn Gly Leu Leu Gln Cys
3815 3820 3825
Met Asn Ser Gly Arg Val Pro Gly Asn Arg Asn Leu Asp Asn Val
3830 3835 3840
Asp Gly Lys Leu Arg Thr Asn Gly Tyr Leu Phe Tyr Pro Asn Arg
3845 3850 3855
Thr Ile Glu Val Pro Arg Val Glu Ala Ala Phe Leu Lys Ser Phe
3860 3865 3870
Gly Phe Gly Gln Ala Gly Ala Glu Val Val Ile Ile His Pro Asp
3875 3880 3885
Arg Leu Leu Ala Val Leu Ser Glu Glu Asn Leu Lys Ser Tyr Ile
3890 3895 3900
Leu Arg Arg Asn Glu Arg Glu Lys Arg Ala Tyr Arg Tyr His Gln
3905 3910 3915
Gly Val Met Ser Gly His His Thr Met Val Gln Val Lys Glu Phe
3920 3925 3930
Ala Pro Tyr Glu Asp Asp Ile Leu Glu Glu Ile Tyr Leu Asn Pro
3935 3940 3945
Arg Ala Arg Ala Ser Phe Asp Ser Ser Lys Ser Thr Trp Thr Phe
3950 3955 3960
His Lys Tyr Arg Asn Asp Lys Asp Thr Val Asp Asp Lys Asp Glu
3965 3970 3975
Met Glu Gln Asp Ser Leu Asn Glu Ser Val Asp Leu Pro Lys Asp
3980 3985 3990
Ile Thr Lys Val Gly Ser Pro Ser Glu Ile Arg Ile Pro Val Lys
3995 4000 4005
Thr Arg Leu Glu Val Thr Val Arg Glu Gly Val Glu Gly Leu Thr
4010 4015 4020
Lys Lys Asp Lys Phe Ser Ser Gln Gly Val Gly Val Asp Val Glu
4025 4030 4035
Pro Val Ser Thr Phe Ala Gln His Glu Glu Lys Thr Ile Phe Ile
4040 4045 4050
Gln Asn Asn Phe Thr Glu Asn Glu Gln Leu Tyr Cys Asn His Ala
4055 4060 4065
Ala Ser Pro Ala Ala Ser Tyr Ala Gly Arg Trp Ala Ala Lys Glu
4070 4075 4080
Ala Val Ile Lys Ala Ile Ser Asn Ser Ser Leu Glu Thr Arg Ser
4085 4090 4095
Leu Trp Gln Gly Ala Glu Gly Lys Leu Ile Asp Ile Glu Ile Ile
4100 4105 4110
Gln Ser Asn Ser Gly Ala Pro Glu Val Val Leu His Gly His Ala
4115 4120 4125
Lys Glu Val Phe Gln Thr Leu Gly Leu Thr Asn Val Lys Val Ser
4130 4135 4140
Ile Ser His Thr Pro Glu Val Ala Val Ala Gln Ala Ile Thr Asn
4145 4150 4155
<210> 13
<211> 181
<212> PRT
<213> 白锈菌
<400> 13
Lys Thr Ile Leu Ile Leu Gly Asp Ser Leu Ser Ala Gly Tyr Gly Ile
1 5 10 15
Asn Pro Glu Gln Gly Trp Val Ala Leu Leu Gln Lys Arg Leu Asp Gln
20 25 30
Gln Phe Pro Lys Gln His Lys Val Ile Asn Ala Ser Val Ser Gly Glu
35 40 45
Thr Thr Ser Gly Ala Leu Ala Arg Leu Pro Lys Leu Leu Thr Thr Tyr
50 55 60
Arg Pro Asn Val Val Val Ile Glu Leu Gly Gly Asn Asp Ala Leu Arg
65 70 75 80
Gly Gln Pro Pro Gln Met Ile Gln Ser Asn Leu Glu Lys Leu Ile Gln
85 90 95
His Ser Gln Lys Ala Lys Ser Lys Val Val Val Phe Gly Met Lys Ile
100 105 110
Pro Pro Asn Tyr Gly Thr Ala Tyr Ser Gln Ala Phe Glu Asn Asn Tyr
115 120 125
Lys Val Val Ser Gln Thr Tyr Gln Val Lys Leu Leu Pro Phe Phe Leu
130 135 140
Asp Gly Val Ala Gly His Lys Ser Leu Met Gln Asn Asp Gln Ile His
145 150 155 160
Pro Asn Ala Lys Ala Gln Ser Ile Leu Leu Asn Asn Ala Tyr Pro Tyr
165 170 175
Ile Lys Gly Ala Leu
180
<210> 14
<211> 1887
<212> PRT
<213> 酿酒酵母
<400> 14
Met Lys Pro Glu Val Glu Gln Glu Leu Ala His Ile Leu Leu Thr Glu
1 5 10 15
Leu Leu Ala Tyr Gln Phe Ala Ser Pro Val Arg Trp Ile Glu Thr Gln
20 25 30
Asp Val Phe Leu Lys Asp Phe Asn Thr Glu Arg Val Val Glu Ile Gly
35 40 45
Pro Ser Pro Thr Leu Ala Gly Met Ala Gln Arg Thr Leu Lys Asn Lys
50 55 60
Tyr Glu Ser Tyr Asp Ala Ala Leu Ser Leu His Arg Glu Ile Leu Cys
65 70 75 80
Tyr Ser Lys Asp Ala Lys Glu Ile Tyr Tyr Thr Pro Asp Pro Ser Glu
85 90 95
Leu Ala Ala Lys Glu Glu Pro Ala Lys Glu Glu Ala Pro Ala Pro Thr
100 105 110
Pro Ala Ala Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Val
115 120 125
Ala Ala Ala Ala Pro Ala Ala Ala Ala Ala Glu Ile Ala Asp Glu Pro
130 135 140
Val Lys Ala Ser Leu Leu Leu His Val Leu Val Ala His Lys Leu Lys
145 150 155 160
Lys Ser Leu Asp Ser Ile Pro Met Ser Lys Thr Ile Lys Asp Leu Val
165 170 175
Gly Gly Lys Ser Thr Val Gln Asn Glu Ile Leu Gly Asp Leu Gly Lys
180 185 190
Glu Phe Gly Thr Thr Pro Glu Lys Pro Glu Glu Thr Pro Leu Glu Glu
195 200 205
Leu Ala Glu Thr Phe Gln Asp Thr Phe Ser Gly Ala Leu Gly Lys Gln
210 215 220
Ser Ser Ser Leu Leu Ser Arg Leu Ile Ser Ser Lys Met Pro Gly Gly
225 230 235 240
Phe Thr Ile Thr Val Ala Arg Lys Tyr Leu Gln Thr Arg Trp Gly Leu
245 250 255
Pro Ser Gly Arg Gln Asp Gly Val Leu Leu Val Ala Leu Ser Asn Glu
260 265 270
Pro Ala Ala Arg Leu Gly Ser Glu Ala Asp Ala Lys Ala Phe Leu Asp
275 280 285
Ser Met Ala Gln Lys Tyr Ala Ser Ile Val Gly Val Asp Leu Ser Ser
290 295 300
Ala Ala Ser Ala Ser Gly Ala Ala Gly Ala Gly Ala Ala Ala Gly Ala
305 310 315 320
Ala Met Ile Asp Ala Gly Ala Leu Glu Glu Ile Thr Lys Asp His Lys
325 330 335
Val Leu Ala Arg Gln Gln Leu Gln Val Leu Ala Arg Tyr Leu Lys Met
340 345 350
Asp Leu Asp Asn Gly Glu Arg Lys Phe Leu Lys Glu Lys Asp Thr Val
355 360 365
Ala Glu Leu Gln Ala Gln Leu Asp Tyr Leu Asn Ala Glu Leu Gly Glu
370 375 380
Phe Phe Val Asn Gly Val Ala Thr Ser Phe Ser Arg Lys Lys Ala Arg
385 390 395 400
Thr Phe Asp Ser Ser Trp Asn Trp Ala Lys Gln Ser Leu Leu Ser Leu
405 410 415
Tyr Phe Glu Ile Ile His Gly Val Leu Lys Asn Val Asp Arg Glu Val
420 425 430
Val Ser Glu Ala Ile Asn Ile Met Asn Arg Ser Asn Asp Ala Leu Ile
435 440 445
Lys Phe Met Glu Tyr His Ile Ser Asn Thr Asp Glu Thr Lys Gly Glu
450 455 460
Asn Tyr Gln Leu Val Lys Thr Leu Gly Glu Gln Leu Ile Glu Asn Cys
465 470 475 480
Lys Gln Val Leu Asp Val Asp Pro Val Tyr Lys Asp Val Ala Lys Pro
485 490 495
Thr Gly Pro Lys Thr Ala Ile Asp Lys Asn Gly Asn Ile Thr Tyr Ser
500 505 510
Glu Glu Pro Arg Glu Lys Val Arg Lys Leu Ser Gln Tyr Val Gln Glu
515 520 525
Met Ala Leu Gly Gly Pro Ile Thr Lys Glu Ser Gln Pro Thr Ile Glu
530 535 540
Glu Asp Leu Thr Arg Val Tyr Lys Ala Ile Ser Ala Gln Ala Asp Lys
545 550 555 560
Gln Asp Ile Ser Ser Ser Thr Arg Val Glu Phe Glu Lys Leu Tyr Ser
565 570 575
Asp Leu Met Lys Phe Leu Glu Ser Ser Lys Glu Ile Asp Pro Ser Gln
580 585 590
Thr Thr Gln Leu Ala Gly Met Asp Val Glu Asp Ala Leu Asp Lys Asp
595 600 605
Ser Thr Lys Glu Val Ala Ser Leu Pro Asn Lys Ser Thr Ile Ser Lys
610 615 620
Thr Val Ser Ser Thr Ile Pro Arg Glu Thr Ile Pro Phe Leu His Leu
625 630 635 640
Arg Lys Lys Thr Pro Ala Gly Asp Trp Lys Tyr Asp Arg Gln Leu Ser
645 650 655
Ser Leu Phe Leu Asp Gly Leu Glu Lys Ala Ala Phe Asn Gly Val Thr
660 665 670
Phe Lys Asp Lys Tyr Val Leu Ile Thr Gly Ala Gly Lys Gly Ser Ile
675 680 685
Gly Ala Glu Val Leu Gln Gly Leu Leu Gln Gly Gly Ala Lys Val Val
690 695 700
Val Thr Thr Ser Arg Phe Ser Lys Gln Val Thr Asp Tyr Tyr Gln Ser
705 710 715 720
Ile Tyr Ala Lys Tyr Gly Ala Lys Gly Ser Thr Leu Ile Val Val Pro
725 730 735
Phe Asn Gln Gly Ser Lys Gln Asp Val Glu Ala Leu Ile Glu Phe Ile
740 745 750
Tyr Asp Thr Glu Lys Asn Gly Gly Leu Gly Trp Asp Leu Asp Ala Ile
755 760 765
Ile Pro Phe Ala Ala Ile Pro Glu Gln Gly Ile Glu Leu Glu His Ile
770 775 780
Asp Ser Lys Ser Glu Phe Ala His Arg Ile Met Leu Thr Asn Ile Leu
785 790 795 800
Arg Met Met Gly Cys Val Lys Lys Gln Lys Ser Ala Arg Gly Ile Glu
805 810 815
Thr Arg Pro Ala Gln Val Ile Leu Pro Met Ser Pro Asn His Gly Thr
820 825 830
Phe Gly Gly Asp Gly Met Tyr Ser Glu Ser Lys Leu Ser Leu Glu Thr
835 840 845
Leu Phe Asn Arg Trp His Ser Glu Ser Trp Ala Asn Gln Leu Thr Val
850 855 860
Cys Gly Ala Ile Ile Gly Trp Thr Arg Gly Thr Gly Leu Met Ser Ala
865 870 875 880
Asn Asn Ile Ile Ala Glu Gly Ile Glu Lys Met Gly Val Arg Thr Phe
885 890 895
Ser Gln Lys Glu Met Ala Phe Asn Leu Leu Gly Leu Leu Thr Pro Glu
900 905 910
Val Val Glu Leu Cys Gln Lys Ser Pro Val Met Ala Asp Leu Asn Gly
915 920 925
Gly Leu Gln Phe Val Pro Glu Leu Lys Glu Phe Thr Ala Lys Leu Arg
930 935 940
Lys Glu Leu Val Glu Thr Ser Glu Val Arg Lys Ala Val Ser Ile Glu
945 950 955 960
Thr Ala Leu Glu His Lys Val Val Asn Gly Asn Ser Ala Asp Ala Ala
965 970 975
Tyr Ala Gln Val Glu Ile Gln Pro Arg Ala Asn Ile Gln Leu Asp Phe
980 985 990
Pro Glu Leu Lys Pro Tyr Lys Gln Val Lys Gln Ile Ala Pro Ala Glu
995 1000 1005
Leu Glu Gly Leu Leu Asp Leu Glu Arg Val Ile Val Val Thr Gly
1010 1015 1020
Phe Ala Glu Val Gly Pro Trp Gly Ser Ala Arg Thr Arg Trp Glu
1025 1030 1035
Met Glu Ala Phe Gly Glu Phe Ser Leu Glu Gly Cys Val Glu Met
1040 1045 1050
Ala Trp Ile Met Gly Phe Ile Ser Tyr His Asn Gly Asn Leu Lys
1055 1060 1065
Gly Arg Pro Tyr Thr Gly Trp Val Asp Ser Lys Thr Lys Glu Pro
1070 1075 1080
Val Asp Asp Lys Asp Val Lys Ala Lys Tyr Glu Thr Ser Ile Leu
1085 1090 1095
Glu His Ser Gly Ile Arg Leu Ile Glu Pro Glu Leu Phe Asn Gly
1100 1105 1110
Tyr Asn Pro Glu Lys Lys Glu Met Ile Gln Glu Val Ile Val Glu
1115 1120 1125
Glu Asp Leu Glu Pro Phe Glu Ala Ser Lys Glu Thr Ala Glu Gln
1130 1135 1140
Phe Lys His Gln His Gly Asp Lys Val Asp Ile Phe Glu Ile Pro
1145 1150 1155
Glu Thr Gly Glu Tyr Ser Val Lys Leu Leu Lys Gly Ala Thr Leu
1160 1165 1170
Tyr Ile Pro Lys Ala Leu Arg Phe Asp Arg Leu Val Ala Gly Gln
1175 1180 1185
Ile Pro Thr Gly Trp Asn Ala Lys Thr Tyr Gly Ile Ser Asp Asp
1190 1195 1200
Ile Ile Ser Gln Val Asp Pro Ile Thr Leu Phe Val Leu Val Ser
1205 1210 1215
Val Val Glu Ala Phe Ile Ala Ser Gly Ile Thr Asp Pro Tyr Glu
1220 1225 1230
Met Tyr Lys Tyr Val His Val Ser Glu Val Gly Asn Cys Ser Gly
1235 1240 1245
Ser Gly Met Gly Gly Val Ser Ala Leu Arg Gly Met Phe Lys Asp
1250 1255 1260
Arg Phe Lys Asp Glu Pro Val Gln Asn Asp Ile Leu Gln Glu Ser
1265 1270 1275
Phe Ile Asn Thr Met Ser Ala Trp Val Asn Met Leu Leu Ile Ser
1280 1285 1290
Ser Ser Gly Pro Ile Lys Thr Pro Val Gly Ala Cys Ala Thr Ser
1295 1300 1305
Val Glu Ser Val Asp Ile Gly Val Glu Thr Ile Leu Ser Gly Lys
1310 1315 1320
Ala Arg Ile Cys Ile Val Gly Gly Tyr Asp Asp Phe Gln Glu Glu
1325 1330 1335
Gly Ser Phe Glu Phe Gly Asn Met Lys Ala Thr Ser Asn Thr Leu
1340 1345 1350
Glu Glu Phe Glu His Gly Arg Thr Pro Ala Glu Met Ser Arg Pro
1355 1360 1365
Ala Thr Thr Thr Arg Asn Gly Phe Met Glu Ala Gln Gly Ala Gly
1370 1375 1380
Ile Gln Ile Ile Met Gln Ala Asp Leu Ala Leu Lys Met Gly Val
1385 1390 1395
Pro Ile Tyr Gly Ile Val Ala Met Ala Ala Thr Ala Thr Asp Lys
1400 1405 1410
Ile Gly Arg Ser Val Pro Ala Pro Gly Lys Gly Ile Leu Thr Thr
1415 1420 1425
Ala Arg Glu His His Ser Ser Val Lys Tyr Ala Ser Pro Asn Leu
1430 1435 1440
Asn Met Lys Tyr Arg Lys Arg Gln Leu Val Thr Arg Glu Ala Gln
1445 1450 1455
Ile Lys Asp Trp Val Glu Asn Glu Leu Glu Ala Leu Lys Leu Glu
1460 1465 1470
Ala Glu Glu Ile Pro Ser Glu Asp Gln Asn Glu Phe Leu Leu Glu
1475 1480 1485
Arg Thr Arg Glu Ile His Asn Glu Ala Glu Ser Gln Leu Arg Ala
1490 1495 1500
Ala Gln Gln Gln Trp Gly Asn Asp Phe Tyr Lys Arg Asp Pro Arg
1505 1510 1515
Ile Ala Pro Leu Arg Gly Ala Leu Ala Thr Tyr Gly Leu Thr Ile
1520 1525 1530
Asp Asp Leu Gly Val Ala Ser Phe His Gly Thr Ser Thr Lys Ala
1535 1540 1545
Asn Asp Lys Asn Glu Ser Ala Thr Ile Asn Glu Met Met Lys His
1550 1555 1560
Leu Gly Arg Ser Glu Gly Asn Pro Val Ile Gly Val Phe Gln Lys
1565 1570 1575
Phe Leu Thr Gly His Pro Lys Gly Ala Ala Gly Ala Trp Met Met
1580 1585 1590
Asn Gly Ala Leu Gln Ile Leu Asn Ser Gly Ile Ile Pro Gly Asn
1595 1600 1605
Arg Asn Ala Asp Asn Val Asp Lys Ile Leu Glu Gln Phe Glu Tyr
1610 1615 1620
Val Leu Tyr Pro Ser Lys Thr Leu Lys Thr Asp Gly Val Arg Ala
1625 1630 1635
Val Ser Ile Thr Ser Phe Gly Phe Gly Gln Lys Gly Gly Gln Ala
1640 1645 1650
Ile Val Val His Pro Asp Tyr Leu Tyr Gly Ala Ile Thr Glu Asp
1655 1660 1665
Arg Tyr Asn Glu Tyr Val Ala Lys Val Ser Ala Arg Glu Lys Ser
1670 1675 1680
Ala Tyr Lys Phe Phe His Asn Gly Met Ile Tyr Asn Lys Leu Phe
1685 1690 1695
Val Ser Lys Glu His Ala Pro Tyr Thr Asp Glu Leu Glu Glu Asp
1700 1705 1710
Val Tyr Leu Asp Pro Leu Ala Arg Val Ser Lys Asp Lys Lys Ser
1715 1720 1725
Gly Ser Leu Thr Phe Asn Ser Lys Asn Ile Gln Ser Lys Asp Ser
1730 1735 1740
Tyr Ile Asn Ala Asn Thr Ile Glu Thr Ala Lys Met Ile Glu Asn
1745 1750 1755
Met Thr Lys Glu Lys Val Ser Asn Gly Gly Val Gly Val Asp Val
1760 1765 1770
Glu Leu Ile Thr Ser Ile Asn Val Glu Asn Asp Thr Phe Ile Glu
1775 1780 1785
Arg Asn Phe Thr Pro Gln Glu Ile Glu Tyr Cys Ser Ala Gln Pro
1790 1795 1800
Ser Val Gln Ser Ser Phe Ala Gly Thr Trp Ser Ala Lys Glu Ala
1805 1810 1815
Val Phe Lys Ser Leu Gly Val Lys Ser Leu Gly Gly Gly Ala Ala
1820 1825 1830
Leu Lys Asp Ile Glu Ile Val Arg Val Asn Lys Asn Ala Pro Ala
1835 1840 1845
Val Glu Leu His Gly Asn Ala Lys Lys Ala Ala Glu Glu Ala Gly
1850 1855 1860
Val Thr Asp Val Lys Val Ser Ile Ser His Asp Asp Leu Gln Ala
1865 1870 1875
Val Ala Val Ala Val Ser Thr Lys Lys
1880 1885
<210> 15
<211> 6285
<212> DNA
<213> 酿酒酵母
<400> 15
atgaagccgg aagttgagca agaattagct catattttgc taactgaatt gttagcttat 60
caatttgcct ctcctgtgag atggattgaa actcaagatg tttttttgaa ggattttaac 120
actgaaaggg ttgttgaaat cggtccttct ccaactttgg ctgggatggc tcaaagaacc 180
ttgaagaata aatacgaatc ttacgatgct gctctgtctt tacatagaga aatcttatgc 240
tattcgaagg atgccaaaga gatttattat accccagatc catccgaact agctgcaaag 300
gaagagcccg ctaaggaaga agctcctgct ccaactccag ctgctagtgc tcctgctcct 360
gcagcagcag ccccagctcc cgtcgcggca gcagccccag ctgcagcagc tgctgagatt 420
aagactatat tgatattggg tgactcattg tccgctggtt atggtattaa tcctgaacaa 480
ggttgggtcg ccttattgca aaagagattg gatcaacaat tcccaaagca acataaagta 540
atcaatgcat ctgtttcagg tgaaactaca tctggtgctt tggcaagatt accaaagttg 600
ttaaccactt acagacctaa cgttgtcgta attgaattgg gtggtaacga cgccttaaga 660
ggtcaaccac ctcaaatgat ccaatcaaat ttggaaaagt taatacaaca ctcccaaaaa 720
gctaagagta aggttgtcgt attcggtatg aagatcccac ctaactatgg tacagcatac 780
tctcaagcct tcgaaaataa ctataaggtt gtctcacaaa cctaccaagt caaattgtta 840
ccatttttct tggatggtgt tgctggtcat aagtccttaa tgcaaaatga ccaaatccac 900
ccaaacgcca aagctcaaag tatattgttg aacaacgctt acccttacat caagggtgca 960
ttagccggcg gtggagctgc tgctgctgcc cccgcgatgg cgttcgccgc tccggccgca 1020
gctggcggtg gagcgcccgc tgccgatgaa cctgtcaagg cttccctatt gttgcacgtt 1080
ttggttgctc acaagttgaa gaagtcgtta gattccattc caatgtccaa gacaatcaaa 1140
gacttggtcg gtggtaaatc tacagtccaa aatgaaattt tgggtgattt aggtaaagaa 1200
tttggtacta ctcctgaaaa accagaagaa actccattag aagaattggc agaaactttc 1260
caagatacct tctctggagc attgggtaag caatcttcct cgttattatc aagattaatc 1320
tcatctaaga tgcctggtgg gtttactatt actgtcgcta gaaaatactt acaaactcgc 1380
tggggactac catctggtag acaagatggt gtccttttgg tagctttatc taacgagcct 1440
gctgctcgtc taggttctga agctgatgcc aaggctttct tggactccat ggctcaaaaa 1500
tacgcttcca ttgttggtgt tgacttatca tcagctgcta gcgctagtgg tgctgccggt 1560
gcaggtgctg ctgccggtgc agctatgatc gatgctggcg ctctggaaga aataaccaaa 1620
gaccacaagg ttttggcgcg tcaacaactg caagtattgg ctcgttatct aaaaatggac 1680
ttggataacg gtgaaagaaa gttcttgaaa gaaaaggaca ctgttgctga acttcaagct 1740
cagttggatt acttgaatgc cgaattaggt gaattctttg ttaacggtgt tgctacttct 1800
ttctctagaa aaaaggccag aaccttcgat tcttcctgga actgggctaa acaatcttta 1860
ttatcattat actttgagat aattcatggt gtcttgaaaa acgttgatag agaggttgtt 1920
agtgaagcta tcaatatcat gaacagatct aacgatgctt tgattaaatt catggaatac 1980
catatctcta acactgatga aacaaaaggt gaaaactatc aattggttaa aactcttggt 2040
gagcagttga ttgaaaactg taaacaagtt ttggatgttg atccagttta caaagatgtt 2100
gctaagccta ccggtccaaa aactgctatt gacaagaacg gtaacattac atactcagaa 2160
gagccaagag aaaaggttag gaaattatct caatacgtac aagaaatggc ccttggtggt 2220
ccaatcacca aagaatctca acctactatt gaagaggatt tgactcgtgt ttacaaggca 2280
atcagtgctc aagctgataa acaagatatt tccagctcca ccagggttga atttgaaaaa 2340
ctatatagtg atttgatgaa gttcttggaa agctccaaag aaatcgatcc ttctcaaaca 2400
acccaattgg ccggtatgga tgttgaggat gctttggaca aagattccac caaagaagtt 2460
gcttctttgc caaacaaatc taccatttct aagacggtat cttcaactat tccaagagaa 2520
actattccgt tcttacattt gagaaagaag actcctgccg gagattggaa atatgaccgc 2580
caattgtctt ctcttttctt agatggttta gaaaaggctg ccttcaacgg tgtcaccttc 2640
aaggacaaat acgtcttgat cactggtgct ggtaagggtt ctattggtgc tgaagtcttg 2700
caaggtttgt tacaaggtgg tgctaaggtt gttgttacca cctctcgttt ctctaagcaa 2760
gttacagact actaccaatc catttacgcc aaatatggtg ctaagggttc tactttgatt 2820
gttgttccat tcaaccaagg ttctaagcaa gacgttgaag ctttgattga atttatctac 2880
gacactgaaa agaatggtgg tttaggttgg gatctagatg ctattattcc attcgcggcc 2940
attccagaac aaggtattga attagaacat attgattcta agtctgaatt tgctcataga 3000
atcatgttga ccaatatctt aagaatgatg ggttgtgtca agaagcaaaa atctgcaaga 3060
ggtattgaaa caagaccagc tcaagtcatt ctaccaatgt ctccaaacca tggtactttc 3120
ggtggtgatg gtatgtattc agaatccaag ttgtctttgg aaactttgtt caacagatgg 3180
cactctgaat cctgggccaa tcaattaacc gtttgcggtg ctattattgg ttggactaga 3240
ggtactggtt taatgagcgc taataacatc attgctgaag gcattgaaaa gatgggtgtt 3300
cgtactttct ctcaaaagga aatggctttc aacttattgg gtctattgac tccagaagtc 3360
gtagaattgt gccaaaaatc acctgttatg gctgacttga atggtggttt gcaatttgtt 3420
cctgaattga aggaattcac tgctaaattg cgtaaagagt tggttgaaac ttctgaagtt 3480
agaaaggcag tttccatcga aactgctttg gagcataagg ttgtcaatgg caatagcgct 3540
gatgctgcat atgctcaagt cgaaattcaa ccaagagcta acattcaact ggacttccca 3600
gaattgaaac catacaaaca ggttaaacaa attgctcccg ctgagcttga aggtttgttg 3660
gatttggaaa gagttattgt agttaccggt tttgctgaag tcggcccatg gggttcggcc 3720
agaacaagat gggaaatgga agcttttggt gaattttcgt tggaaggttg cgttgaaatg 3780
gcctggatta tgggcttcat ttcataccat aacggtaatt tgaagggtcg tccatacact 3840
ggttgggttg attccaaaac aaaagaacca gttgatgaca aggacgttaa ggccaagtat 3900
gaaacatcaa tcctagaaca cagtggtatc agattgatcg aaccagagtt attcaatggt 3960
tacaacccag aaaagaagga aatgattcaa gaagtcattg tcgaagaaga cttggaacca 4020
tttgaggctt cgaaggaaac tgccgaacaa tttaaacacc aacatggtga caaagtggat 4080
atcttcgaaa tcccagaaac aggagagtac tctgttaagt tactaaaggg tgccacttta 4140
tacattccaa aggctttgag atttgaccgt ttggttgcag gtcaaattcc aactggttgg 4200
aatgctaaga cttatggtat ctctgatgat atcatttctc aggttgaccc aatcacatta 4260
ttcgttttgg tctctgttgt ggaagcattt attgcatctg gtatcaccga cccatacgaa 4320
atgtacaaat acgtacatgt ttctgaggtt ggtaactgtt ctggttctgg tatgggtggt 4380
gtttctgcct tacgtggtat gtttaaggac cgtttcaagg atgagcctgt ccaaaatgat 4440
attttacaag aatcatttat caacaccatg tccgcttggg ttaatatgtt gttgatttcc 4500
tcatctggtc caatcaagac acctgttggt gcctgtgcca catccgtgga atctgttgac 4560
attggtgtag aaaccatctt gtctggtaag gctagaatct gtattgtcgg tggttacgat 4620
gatttccaag aagaaggctc ctttgagttc ggtaacatga aggccacttc caacactttg 4680
gaagaatttg aacatggtcg taccccagcg gaaatgtcca gacctgccac cactacccgt 4740
aacggtttta tggaagctca aggtgctggt attcaaatca tcatgcaagc tgatttagct 4800
ttgaagatgg gtgtgccaat ttacggtatt gttgccatgg ctgctaccgc caccgataag 4860
attggtagat ctgtgccagc tccaggtaag ggtattttaa ccactgctcg tgaacaccac 4920
tccagtgtta agtatgcttc accaaacttg aacatgaagt acagaaagcg ccaattggtt 4980
actcgtgaag ctcagattaa agattgggta gaaaacgaat tggaagcttt gaagttggag 5040
gccgaagaaa ttccaagcga agaccaaaac gagttcttac ttgaacgtac cagagaaatc 5100
cacaacgaag ctgaaagtca attgagagct gcacaacaac aatggggtaa cgacttctac 5160
aagagggacc cacgtattgc tccattgaga ggagcactgg ctacttacgg tttaactatt 5220
gatgacttgg gtgtcgcttc attccacggt acatccacaa aggctaatga caagaacgaa 5280
tctgccacaa ttaatgaaat gatgaagcat ttgggtagat ctgaaggtaa tcccgtcatt 5340
ggtgttttcc aaaagttctt gactggtcat ccaaagggtg ctgctggtgc atggatgatg 5400
aatggtgctt tgcaaattct aaacagtggt attattccag gtaaccgtaa cgctgataac 5460
gtggataaga tcttggagca atttgaatac gtcttgtacc catccaagac tttaaagacc 5520
gacggtgtca gagccgtgtc catcacttct ttcggttttg gtcaaaaggg tggtcaagct 5580
attgtggttc atccagacta cttatacggt gctatcactg aagacagata caacgagtat 5640
gtcgccaagg ttagtgccag agagaaaagt gcctacaaat tcttccataa tggtatgatc 5700
tacaacaagt tgttcgtaag taaagagcat gctccataca ctgatgaatt ggaagaggat 5760
gtttacttgg acccattagc ccgtgtatct aaggataaga aatcaggctc cttgactttc 5820
aactctaaaa acatccaaag caaggacagt tacatcaatg ctaacaccat tgaaactgcc 5880
aagatgattg aaaacatgac caaggagaaa gtctctaacg gtggcgtcgg tgtagatgtt 5940
gaattaatca ctagcatcaa cgttgaaaat gatactttta tcgagcgcaa tttcaccccg 6000
caagaaatag agtactgcag cgcgcagcct agtgtgcaaa gctctttcgc tgggacatgg 6060
tccgccaaag aggctgtttt caagtcctta ggcgtcaagt ccttaggcgg tggtgctgca 6120
ttgaaagaca tcgaaatcgt acgcgttaac aaaaacgctc cagccgttga actgcacggt 6180
aacgccaaaa aggctgccga agaagctggt gttaccgatg tgaaggtatc tatttctcac 6240
gatgacctcc aagctgtcgc ggtcgccgtt tctactaaga aatag 6285
<210> 16
<211> 6291
<212> DNA
<213> 酿酒酵母
<400> 16
atgaagccgg aagttgagca agaattagct catattttgc taactgaatt gttagcttat 60
caatttgcct ctcctgtgag atggattgaa actcaagatg tttttttgaa ggattttaac 120
actgaaaggg ttgttgaaat cggtccttct ccaactttgg ctgggatggc tcaaagaacc 180
ttgaagaata aatacgaatc ttacgatgct gctctgtctt tacatagaga aatcttatgc 240
tattcgaagg atgccaaaga gatttattat accccagatc catccgaact agctgcaaag 300
gaagagcccg ctaaggaaga agctcctgct ccaactccag ctgctagtgc tcctgctcct 360
gcagcagcag ccccagctcc cgtcgcggca gcagccccag ctgcagcagc tgctgagatt 420
gccgatgaac ctgtcaaggc ttccctattg ttgcacgttt tggttgctca caagttgaag 480
aagtcgttag attccattcc aatgtccaag acaatcaaag acttggtcgg tggtaaatct 540
acagtccaaa atgaaatttt gggtgattta ggtaaagaat ttggtactac tcctgaaaaa 600
ccagaagaaa ctccattaga agaattggca gaaactttcc aagatacctt ctctggagca 660
ttgggtaagc aatcttcctc gttattatca agattaatct catctaagat gcctggtggg 720
tttactatta ctgtcgctag aaaatactta caaactcgct ggggactacc atctggtaga 780
caagatggtg tccttttggt agctttatct aacgagcctg ctgctcgtct aggttctgaa 840
gctgatgcca aggctttctt ggactccatg gctcaaaaat acgcttccat tgttggtgtt 900
gacttatcat cagctgccgg cggtggagct gctgctgctg cccccgcgat ggcgttcgcc 960
gctccggccg cagctggcgg tggagcgccc gctaagacta tattgatatt gggtgactca 1020
ttgtccgctg gttatggtat taatcctgaa caaggttggg tcgccttatt gcaaaagaga 1080
ttggatcaac aattcccaaa gcaacataaa gtaatcaatg catctgtttc aggtgaaact 1140
acatctggtg ctttggcaag attaccaaag ttgttaacca cttacagacc taacgttgtc 1200
gtaattgaat tgggtggtaa cgacgcctta agaggtcaac cacctcaaat gatccaatca 1260
aatttggaaa agttaataca acactcccaa aaagctaaga gtaaggttgt cgtattcggt 1320
atgaagatcc cacctaacta tggtacagca tactctcaag ccttcgaaaa taactataag 1380
gttgtctcac aaacctacca agtcaaattg ttaccatttt tcttggatgg tgttgctggt 1440
cataagtcct taatgcaaaa tgaccaaatc cacccaaacg ccaaagctca aagtatattg 1500
ttgaacaacg cttaccctta catcaagggt gcattagctg gtgctagcgc tagtggtgct 1560
gccggtgcag gtgctgctgc cggtgcagct atgatcgatg ctggcgctct ggaagaaata 1620
accaaagacc acaaggtttt ggcgcgtcaa caactgcaag tattggctcg ttatctaaaa 1680
atggacttgg ataacggtga aagaaagttc ttgaaagaaa aggacactgt tgctgaactt 1740
caagctcagt tggattactt gaatgccgaa ttaggtgaat tctttgttaa cggtgttgct 1800
acttctttct ctagaaaaaa ggccagaacc ttcgattctt cctggaactg ggctaaacaa 1860
tctttattat cattatactt tgagataatt catggtgtct tgaaaaacgt tgatagagag 1920
gttgttagtg aagctatcaa tatcatgaac agatctaacg atgctttgat taaattcatg 1980
gaataccata tctctaacac tgatgaaaca aaaggtgaaa actatcaatt ggttaaaact 2040
cttggtgagc agttgattga aaactgtaaa caagttttgg atgttgatcc agtttacaaa 2100
gatgttgcta agcctaccgg tccaaaaact gctattgaca agaacggtaa cattacatac 2160
tcagaagagc caagagaaaa ggttaggaaa ttatctcaat acgtacaaga aatggccctt 2220
ggtggtccaa tcaccaaaga atctcaacct actattgaag aggatttgac tcgtgtttac 2280
aaggcaatca gtgctcaagc tgataaacaa gatatttcca gctccaccag ggttgaattt 2340
gaaaaactat atagtgattt gatgaagttc ttggaaagct ccaaagaaat cgatccttct 2400
caaacaaccc aattggccgg tatggatgtt gaggatgctt tggacaaaga ttccaccaaa 2460
gaagttgctt ctttgccaaa caaatctacc atttctaaga cggtatcttc aactattcca 2520
agagaaacta ttccgttctt acatttgaga aagaagactc ctgccggaga ttggaaatat 2580
gaccgccaat tgtcttctct tttcttagat ggtttagaaa aggctgcctt caacggtgtc 2640
accttcaagg acaaatacgt cttgatcact ggtgctggta agggttctat tggtgctgaa 2700
gtcttgcaag gtttgttaca aggtggtgct aaggttgttg ttaccacctc tcgtttctct 2760
aagcaagtta cagactacta ccaatccatt tacgccaaat atggtgctaa gggttctact 2820
ttgattgttg ttccattcaa ccaaggttct aagcaagacg ttgaagcttt gattgaattt 2880
atctacgaca ctgaaaagaa tggtggttta ggttgggatc tagatgctat tattccattc 2940
gcggccattc cagaacaagg tattgaatta gaacatattg attctaagtc tgaatttgct 3000
catagaatca tgttgaccaa tatcttaaga atgatgggtt gtgtcaagaa gcaaaaatct 3060
gcaagaggta ttgaaacaag accagctcaa gtcattctac caatgtctcc aaaccatggt 3120
actttcggtg gtgatggtat gtattcagaa tccaagttgt ctttggaaac tttgttcaac 3180
agatggcact ctgaatcctg ggccaatcaa ttaaccgttt gcggtgctat tattggttgg 3240
actagaggta ctggtttaat gagcgctaat aacatcattg ctgaaggcat tgaaaagatg 3300
ggtgttcgta ctttctctca aaaggaaatg gctttcaact tattgggtct attgactcca 3360
gaagtcgtag aattgtgcca aaaatcacct gttatggctg acttgaatgg tggtttgcaa 3420
tttgttcctg aattgaagga attcactgct aaattgcgta aagagttggt tgaaacttct 3480
gaagttagaa aggcagtttc catcgaaact gctttggagc ataaggttgt caatggcaat 3540
agcgctgatg ctgcatatgc tcaagtcgaa attcaaccaa gagctaacat tcaactggac 3600
ttcccagaat tgaaaccata caaacaggtt aaacaaattg ctcccgctga gcttgaaggt 3660
ttgttggatt tggaaagagt tattgtagtt accggttttg ctgaagtcgg cccatggggt 3720
tcggccagaa caagatggga aatggaagct tttggtgaat tttcgttgga aggttgcgtt 3780
gaaatggcct ggattatggg cttcatttca taccataacg gtaatttgaa gggtcgtcca 3840
tacactggtt gggttgattc caaaacaaaa gaaccagttg atgacaagga cgttaaggcc 3900
aagtatgaaa catcaatcct agaacacagt ggtatcagat tgatcgaacc agagttattc 3960
aatggttaca acccagaaaa gaaggaaatg attcaagaag tcattgtcga agaagacttg 4020
gaaccatttg aggcttcgaa ggaaactgcc gaacaattta aacaccaaca tggtgacaaa 4080
gtggatatct tcgaaatccc agaaacagga gagtactctg ttaagttact aaagggtgcc 4140
actttataca ttccaaaggc tttgagattt gaccgtttgg ttgcaggtca aattccaact 4200
ggttggaatg ctaagactta tggtatctct gatgatatca tttctcaggt tgacccaatc 4260
acattattcg ttttggtctc tgttgtggaa gcatttattg catctggtat caccgaccca 4320
tacgaaatgt acaaatacgt acatgtttct gaggttggta actgttctgg ttctggtatg 4380
ggtggtgttt ctgccttacg tggtatgttt aaggaccgtt tcaaggatga gcctgtccaa 4440
aatgatattt tacaagaatc atttatcaac accatgtccg cttgggttaa tatgttgttg 4500
atttcctcat ctggtccaat caagacacct gttggtgcct gtgccacatc cgtggaatct 4560
gttgacattg gtgtagaaac catcttgtct ggtaaggcta gaatctgtat tgtcggtggt 4620
tacgatgatt tccaagaaga aggctccttt gagttcggta acatgaaggc cacttccaac 4680
actttggaag aatttgaaca tggtcgtacc ccagcggaaa tgtccagacc tgccaccact 4740
acccgtaacg gttttatgga agctcaaggt gctggtattc aaatcatcat gcaagctgat 4800
ttagctttga agatgggtgt gccaatttac ggtattgttg ccatggctgc taccgccacc 4860
gataagattg gtagatctgt gccagctcca ggtaagggta ttttaaccac tgctcgtgaa 4920
caccactcca gtgttaagta tgcttcacca aacttgaaca tgaagtacag aaagcgccaa 4980
ttggttactc gtgaagctca gattaaagat tgggtagaaa acgaattgga agctttgaag 5040
ttggaggccg aagaaattcc aagcgaagac caaaacgagt tcttacttga acgtaccaga 5100
gaaatccaca acgaagctga aagtcaattg agagctgcac aacaacaatg gggtaacgac 5160
ttctacaaga gggacccacg tattgctcca ttgagaggag cactggctac ttacggttta 5220
actattgatg acttgggtgt cgcttcattc cacggtacat ccacaaaggc taatgacaag 5280
aacgaatctg ccacaattaa tgaaatgatg aagcatttgg gtagatctga aggtaatccc 5340
gtcattggtg ttttccaaaa gttcttgact ggtcatccaa agggtgctgc tggtgcatgg 5400
atgatgaatg gtgctttgca aattctaaac agtggtatta ttccaggtaa ccgtaacgct 5460
gataacgtgg ataagatctt ggagcaattt gaatacgtct tgtacccatc caagacttta 5520
aagaccgacg gtgtcagagc cgtgtccatc acttctttcg gttttggtca aaagggtggt 5580
caagctattg tggttcatcc agactactta tacggtgcta tcactgaaga cagatacaac 5640
gagtatgtcg ccaaggttag tgccagagag aaaagtgcct acaaattctt ccataatggt 5700
atgatctaca acaagttgtt cgtaagtaaa gagcatgctc catacactga tgaattggaa 5760
gaggatgttt acttggaccc attagcccgt gtatctaagg ataagaaatc aggctccttg 5820
actttcaact ctaaaaacat ccaaagcaag gacagttaca tcaatgctaa caccattgaa 5880
actgccaaga tgattgaaaa catgaccaag gagaaagtct ctaacggtgg cgtcggtgta 5940
gatgttgaat taatcactag catcaacgtt gaaaatgata cttttatcga gcgcaatttc 6000
accccgcaag aaatagagta ctgcagcgcg cagcctagtg tgcaaagctc tttcgctggg 6060
acatggtccg ccaaagaggc tgttttcaag tccttaggcg tcaagtcctt aggcggtggt 6120
gctgcattga aagacatcga aatcgtacgc gttaacaaaa acgctccagc cgttgaactg 6180
cacggtaacg ccaaaaaggc tgccgaagaa gctggtgtta ccgatgtgaa ggtatctatt 6240
tctcacgatg acctccaagc tgtcgcggtc gccgtttcta ctaagaaata g 6291
<210> 17
<211> 2039
<212> PRT
<213> 酿酒酵母
<400> 17
Met Ala Gln Arg Thr Leu Lys Asn Lys Tyr Glu Ser Tyr Asp Ala Ala
1 5 10 15
Leu Ser Leu His Arg Glu Ile Leu Cys Tyr Ser Lys Asp Ala Lys Glu
20 25 30
Ile Tyr Tyr Thr Pro Asp Pro Ser Glu Leu Ala Ala Lys Glu Glu Pro
35 40 45
Ala Lys Glu Glu Ala Pro Ala Pro Thr Pro Ala Ala Ser Ala Pro Ala
50 55 60
Pro Ala Ala Ala Ala Pro Ala Pro Val Ala Ala Ala Ala Pro Ala Ala
65 70 75 80
Ala Ala Ala Glu Ile Lys Thr Ile Leu Ile Leu Gly Asp Ser Leu Ser
85 90 95
Ala Gly Tyr Gly Ile Asn Pro Glu Gln Gly Trp Val Ala Leu Leu Gln
100 105 110
Lys Arg Leu Asp Gln Gln Phe Pro Lys Gln His Lys Val Ile Asn Ala
115 120 125
Ser Val Ser Gly Glu Thr Thr Ser Gly Ala Leu Ala Arg Leu Pro Lys
130 135 140
Leu Leu Thr Thr Tyr Arg Pro Asn Val Val Val Ile Glu Leu Gly Gly
145 150 155 160
Asn Asp Ala Leu Arg Gly Gln Pro Pro Gln Met Ile Gln Ser Asn Leu
165 170 175
Glu Lys Leu Ile Gln His Ser Gln Lys Ala Lys Ser Lys Val Val Val
180 185 190
Phe Gly Met Lys Ile Pro Pro Asn Tyr Gly Thr Ala Tyr Ser Gln Ala
195 200 205
Phe Glu Asn Asn Tyr Lys Val Val Ser Gln Thr Tyr Gln Val Lys Leu
210 215 220
Leu Pro Phe Phe Leu Asp Gly Val Ala Gly His Lys Ser Leu Met Gln
225 230 235 240
Asn Asp Gln Ile His Pro Asn Ala Lys Ala Gln Ser Ile Leu Leu Asn
245 250 255
Asn Ala Tyr Pro Tyr Ile Lys Gly Ala Leu Ala Gly Gly Gly Ala Ala
260 265 270
Ala Ala Ala Pro Ala Met Ala Phe Ala Ala Pro Ala Ala Ala Gly Gly
275 280 285
Gly Ala Pro Ala Ala Asp Glu Pro Val Lys Ala Ser Leu Leu Leu His
290 295 300
Val Leu Val Ala His Lys Leu Lys Lys Ser Leu Asp Ser Ile Pro Met
305 310 315 320
Ser Lys Thr Ile Lys Asp Leu Val Gly Gly Lys Ser Thr Val Gln Asn
325 330 335
Glu Ile Leu Gly Asp Leu Gly Lys Glu Phe Gly Thr Thr Pro Glu Lys
340 345 350
Pro Glu Glu Thr Pro Leu Glu Glu Leu Ala Glu Thr Phe Gln Asp Thr
355 360 365
Phe Ser Gly Ala Leu Gly Lys Gln Ser Ser Ser Leu Leu Ser Arg Leu
370 375 380
Ile Ser Ser Lys Met Pro Gly Gly Phe Thr Ile Thr Val Ala Arg Lys
385 390 395 400
Tyr Leu Gln Thr Arg Trp Gly Leu Pro Ser Gly Arg Gln Asp Gly Val
405 410 415
Leu Leu Val Ala Leu Ser Asn Glu Pro Ala Ala Arg Leu Gly Ser Glu
420 425 430
Ala Asp Ala Lys Ala Phe Leu Asp Ser Met Ala Gln Lys Tyr Ala Ser
435 440 445
Ile Val Gly Val Asp Leu Ser Ser Ala Ala Ser Ala Ser Gly Ala Ala
450 455 460
Gly Ala Gly Ala Ala Ala Gly Ala Ala Met Ile Asp Ala Gly Ala Leu
465 470 475 480
Glu Glu Ile Thr Lys Asp His Lys Val Leu Ala Arg Gln Gln Leu Gln
485 490 495
Val Leu Ala Arg Tyr Leu Lys Met Asp Leu Asp Asn Gly Glu Arg Lys
500 505 510
Phe Leu Lys Glu Lys Asp Thr Val Ala Glu Leu Gln Ala Gln Leu Asp
515 520 525
Tyr Leu Asn Ala Glu Leu Gly Glu Phe Phe Val Asn Gly Val Ala Thr
530 535 540
Ser Phe Ser Arg Lys Lys Ala Arg Thr Phe Asp Ser Ser Trp Asn Trp
545 550 555 560
Ala Lys Gln Ser Leu Leu Ser Leu Tyr Phe Glu Ile Ile His Gly Val
565 570 575
Leu Lys Asn Val Asp Arg Glu Val Val Ser Glu Ala Ile Asn Ile Met
580 585 590
Asn Arg Ser Asn Asp Ala Leu Ile Lys Phe Met Glu Tyr His Ile Ser
595 600 605
Asn Thr Asp Glu Thr Lys Gly Glu Asn Tyr Gln Leu Val Lys Thr Leu
610 615 620
Gly Glu Gln Leu Ile Glu Asn Cys Lys Gln Val Leu Asp Val Asp Pro
625 630 635 640
Val Tyr Lys Asp Val Ala Lys Pro Thr Gly Pro Lys Thr Ala Ile Asp
645 650 655
Lys Asn Gly Asn Ile Thr Tyr Ser Glu Glu Pro Arg Glu Lys Val Arg
660 665 670
Lys Leu Ser Gln Tyr Val Gln Glu Met Ala Leu Gly Gly Pro Ile Thr
675 680 685
Lys Glu Ser Gln Pro Thr Ile Glu Glu Asp Leu Thr Arg Val Tyr Lys
690 695 700
Ala Ile Ser Ala Gln Ala Asp Lys Gln Asp Ile Ser Ser Ser Thr Arg
705 710 715 720
Val Glu Phe Glu Lys Leu Tyr Ser Asp Leu Met Lys Phe Leu Glu Ser
725 730 735
Ser Lys Glu Ile Asp Pro Ser Gln Thr Thr Gln Leu Ala Gly Met Asp
740 745 750
Val Glu Asp Ala Leu Asp Lys Asp Ser Thr Lys Glu Val Ala Ser Leu
755 760 765
Pro Asn Lys Ser Thr Ile Ser Lys Thr Val Ser Ser Thr Ile Pro Arg
770 775 780
Glu Thr Ile Pro Phe Leu His Leu Arg Lys Lys Thr Pro Ala Gly Asp
785 790 795 800
Trp Lys Tyr Asp Arg Gln Leu Ser Ser Leu Phe Leu Asp Gly Leu Glu
805 810 815
Lys Ala Ala Phe Asn Gly Val Thr Phe Lys Asp Lys Tyr Val Leu Ile
820 825 830
Thr Gly Ala Gly Lys Gly Ser Ile Gly Ala Glu Val Leu Gln Gly Leu
835 840 845
Leu Gln Gly Gly Ala Lys Val Val Val Thr Thr Ser Arg Phe Ser Lys
850 855 860
Gln Val Thr Asp Tyr Tyr Gln Ser Ile Tyr Ala Lys Tyr Gly Ala Lys
865 870 875 880
Gly Ser Thr Leu Ile Val Val Pro Phe Asn Gln Gly Ser Lys Gln Asp
885 890 895
Val Glu Ala Leu Ile Glu Phe Ile Tyr Asp Thr Glu Lys Asn Gly Gly
900 905 910
Leu Gly Trp Asp Leu Asp Ala Ile Ile Pro Phe Ala Ala Ile Pro Glu
915 920 925
Gln Gly Ile Glu Leu Glu His Ile Asp Ser Lys Ser Glu Phe Ala His
930 935 940
Arg Ile Met Leu Thr Asn Ile Leu Arg Met Met Gly Cys Val Lys Lys
945 950 955 960
Gln Lys Ser Ala Arg Gly Ile Glu Thr Arg Pro Ala Gln Val Ile Leu
965 970 975
Pro Met Ser Pro Asn His Gly Thr Phe Gly Gly Asp Gly Met Tyr Ser
980 985 990
Glu Ser Lys Leu Ser Leu Glu Thr Leu Phe Asn Arg Trp His Ser Glu
995 1000 1005
Ser Trp Ala Asn Gln Leu Thr Val Cys Gly Ala Ile Ile Gly Trp
1010 1015 1020
Thr Arg Gly Thr Gly Leu Met Ser Ala Asn Asn Ile Ile Ala Glu
1025 1030 1035
Gly Ile Glu Lys Met Gly Val Arg Thr Phe Ser Gln Lys Glu Met
1040 1045 1050
Ala Phe Asn Leu Leu Gly Leu Leu Thr Pro Glu Val Val Glu Leu
1055 1060 1065
Cys Gln Lys Ser Pro Val Met Ala Asp Leu Asn Gly Gly Leu Gln
1070 1075 1080
Phe Val Pro Glu Leu Lys Glu Phe Thr Ala Lys Leu Arg Lys Glu
1085 1090 1095
Leu Val Glu Thr Ser Glu Val Arg Lys Ala Val Ser Ile Glu Thr
1100 1105 1110
Ala Leu Glu His Lys Val Val Asn Gly Asn Ser Ala Asp Ala Ala
1115 1120 1125
Tyr Ala Gln Val Glu Ile Gln Pro Arg Ala Asn Ile Gln Leu Asp
1130 1135 1140
Phe Pro Glu Leu Lys Pro Tyr Lys Gln Val Lys Gln Ile Ala Pro
1145 1150 1155
Ala Glu Leu Glu Gly Leu Leu Asp Leu Glu Arg Val Ile Val Val
1160 1165 1170
Thr Gly Phe Ala Glu Val Gly Pro Trp Gly Ser Ala Arg Thr Arg
1175 1180 1185
Trp Glu Met Glu Ala Phe Gly Glu Phe Ser Leu Glu Gly Cys Val
1190 1195 1200
Glu Met Ala Trp Ile Met Gly Phe Ile Ser Tyr His Asn Gly Asn
1205 1210 1215
Leu Lys Gly Arg Pro Tyr Thr Gly Trp Val Asp Ser Lys Thr Lys
1220 1225 1230
Glu Pro Val Asp Asp Lys Asp Val Lys Ala Lys Tyr Glu Thr Ser
1235 1240 1245
Ile Leu Glu His Ser Gly Ile Arg Leu Ile Glu Pro Glu Leu Phe
1250 1255 1260
Asn Gly Tyr Asn Pro Glu Lys Lys Glu Met Ile Gln Glu Val Ile
1265 1270 1275
Val Glu Glu Asp Leu Glu Pro Phe Glu Ala Ser Lys Glu Thr Ala
1280 1285 1290
Glu Gln Phe Lys His Gln His Gly Asp Lys Val Asp Ile Phe Glu
1295 1300 1305
Ile Pro Glu Thr Gly Glu Tyr Ser Val Lys Leu Leu Lys Gly Ala
1310 1315 1320
Thr Leu Tyr Ile Pro Lys Ala Leu Arg Phe Asp Arg Leu Val Ala
1325 1330 1335
Gly Gln Ile Pro Thr Gly Trp Asn Ala Lys Thr Tyr Gly Ile Ser
1340 1345 1350
Asp Asp Ile Ile Ser Gln Val Asp Pro Ile Thr Leu Phe Val Leu
1355 1360 1365
Val Ser Val Val Glu Ala Phe Ile Ala Ser Gly Ile Thr Asp Pro
1370 1375 1380
Tyr Glu Met Tyr Lys Tyr Val His Val Ser Glu Val Gly Asn Cys
1385 1390 1395
Ser Gly Ser Gly Met Gly Gly Val Ser Ala Leu Arg Gly Met Phe
1400 1405 1410
Lys Asp Arg Phe Lys Asp Glu Pro Val Gln Asn Asp Ile Leu Gln
1415 1420 1425
Glu Ser Phe Ile Asn Thr Met Ser Ala Trp Val Asn Met Leu Leu
1430 1435 1440
Ile Ser Ser Ser Gly Pro Ile Lys Thr Pro Val Gly Ala Cys Ala
1445 1450 1455
Thr Ser Val Glu Ser Val Asp Ile Gly Val Glu Thr Ile Leu Ser
1460 1465 1470
Gly Lys Ala Arg Ile Cys Ile Val Gly Gly Tyr Asp Asp Phe Gln
1475 1480 1485
Glu Glu Gly Ser Phe Glu Phe Gly Asn Met Lys Ala Thr Ser Asn
1490 1495 1500
Thr Leu Glu Glu Phe Glu His Gly Arg Thr Pro Ala Glu Met Ser
1505 1510 1515
Arg Pro Ala Thr Thr Thr Arg Asn Gly Phe Met Glu Ala Gln Gly
1520 1525 1530
Ala Gly Ile Gln Ile Ile Met Gln Ala Asp Leu Ala Leu Lys Met
1535 1540 1545
Gly Val Pro Ile Tyr Gly Ile Val Ala Met Ala Ala Thr Ala Thr
1550 1555 1560
Asp Lys Ile Gly Arg Ser Val Pro Ala Pro Gly Lys Gly Ile Leu
1565 1570 1575
Thr Thr Ala Arg Glu His His Ser Ser Val Lys Tyr Ala Ser Pro
1580 1585 1590
Asn Leu Asn Met Lys Tyr Arg Lys Arg Gln Leu Val Thr Arg Glu
1595 1600 1605
Ala Gln Ile Lys Asp Trp Val Glu Asn Glu Leu Glu Ala Leu Lys
1610 1615 1620
Leu Glu Ala Glu Glu Ile Pro Ser Glu Asp Gln Asn Glu Phe Leu
1625 1630 1635
Leu Glu Arg Thr Arg Glu Ile His Asn Glu Ala Glu Ser Gln Leu
1640 1645 1650
Arg Ala Ala Gln Gln Gln Trp Gly Asn Asp Phe Tyr Lys Arg Asp
1655 1660 1665
Pro Arg Ile Ala Pro Leu Arg Gly Ala Leu Ala Thr Tyr Gly Leu
1670 1675 1680
Thr Ile Asp Asp Leu Gly Val Ala Ser Phe His Gly Thr Ser Thr
1685 1690 1695
Lys Ala Asn Asp Lys Asn Glu Ser Ala Thr Ile Asn Glu Met Met
1700 1705 1710
Lys His Leu Gly Arg Ser Glu Gly Asn Pro Val Ile Gly Val Phe
1715 1720 1725
Gln Lys Phe Leu Thr Gly His Pro Lys Gly Ala Ala Gly Ala Trp
1730 1735 1740
Met Met Asn Gly Ala Leu Gln Ile Leu Asn Ser Gly Ile Ile Pro
1745 1750 1755
Gly Asn Arg Asn Ala Asp Asn Val Asp Lys Ile Leu Glu Gln Phe
1760 1765 1770
Glu Tyr Val Leu Tyr Pro Ser Lys Thr Leu Lys Thr Asp Gly Val
1775 1780 1785
Arg Ala Val Ser Ile Thr Ser Phe Gly Phe Gly Gln Lys Gly Gly
1790 1795 1800
Gln Ala Ile Val Val His Pro Asp Tyr Leu Tyr Gly Ala Ile Thr
1805 1810 1815
Glu Asp Arg Tyr Asn Glu Tyr Val Ala Lys Val Ser Ala Arg Glu
1820 1825 1830
Lys Ser Ala Tyr Lys Phe Phe His Asn Gly Met Ile Tyr Asn Lys
1835 1840 1845
Leu Phe Val Ser Lys Glu His Ala Pro Tyr Thr Asp Glu Leu Glu
1850 1855 1860
Glu Asp Val Tyr Leu Asp Pro Leu Ala Arg Val Ser Lys Asp Lys
1865 1870 1875
Lys Ser Gly Ser Leu Thr Phe Asn Ser Lys Asn Ile Gln Ser Lys
1880 1885 1890
Asp Ser Tyr Ile Asn Ala Asn Thr Ile Glu Thr Ala Lys Met Ile
1895 1900 1905
Glu Asn Met Thr Lys Glu Lys Val Ser Asn Gly Gly Val Gly Val
1910 1915 1920
Asp Val Glu Leu Ile Thr Ser Ile Asn Val Glu Asn Asp Thr Phe
1925 1930 1935
Ile Glu Arg Asn Phe Thr Pro Gln Glu Ile Glu Tyr Cys Ser Ala
1940 1945 1950
Gln Pro Ser Val Gln Ser Ser Phe Ala Gly Thr Trp Ser Ala Lys
1955 1960 1965
Glu Ala Val Phe Lys Ser Leu Gly Val Lys Ser Leu Gly Gly Gly
1970 1975 1980
Ala Ala Leu Lys Asp Ile Glu Ile Val Arg Val Asn Lys Asn Ala
1985 1990 1995
Pro Ala Val Glu Leu His Gly Asn Ala Lys Lys Ala Ala Glu Glu
2000 2005 2010
Ala Gly Val Thr Asp Val Lys Val Ser Ile Ser His Asp Asp Leu
2015 2020 2025
Gln Ala Val Ala Val Ala Val Ser Thr Lys Lys
2030 2035
<210> 18
<211> 2096
<212> PRT
<213> 酿酒酵母
<400> 18
Met Lys Pro Glu Val Glu Gln Glu Leu Ala His Ile Leu Leu Thr Glu
1 5 10 15
Leu Leu Ala Tyr Gln Phe Ala Ser Pro Val Arg Trp Ile Glu Thr Gln
20 25 30
Asp Val Phe Leu Lys Asp Phe Asn Thr Glu Arg Val Val Glu Ile Gly
35 40 45
Pro Ser Pro Thr Leu Ala Gly Met Ala Gln Arg Thr Leu Lys Asn Lys
50 55 60
Tyr Glu Ser Tyr Asp Ala Ala Leu Ser Leu His Arg Glu Ile Leu Cys
65 70 75 80
Tyr Ser Lys Asp Ala Lys Glu Ile Tyr Tyr Thr Pro Asp Pro Ser Glu
85 90 95
Leu Ala Ala Lys Glu Glu Pro Ala Lys Glu Glu Ala Pro Ala Pro Thr
100 105 110
Pro Ala Ala Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Val
115 120 125
Ala Ala Ala Ala Pro Ala Ala Ala Ala Ala Glu Ile Ala Asp Glu Pro
130 135 140
Val Lys Ala Ser Leu Leu Leu His Val Leu Val Ala His Lys Leu Lys
145 150 155 160
Lys Ser Leu Asp Ser Ile Pro Met Ser Lys Thr Ile Lys Asp Leu Val
165 170 175
Gly Gly Lys Ser Thr Val Gln Asn Glu Ile Leu Gly Asp Leu Gly Lys
180 185 190
Glu Phe Gly Thr Thr Pro Glu Lys Pro Glu Glu Thr Pro Leu Glu Glu
195 200 205
Leu Ala Glu Thr Phe Gln Asp Thr Phe Ser Gly Ala Leu Gly Lys Gln
210 215 220
Ser Ser Ser Leu Leu Ser Arg Leu Ile Ser Ser Lys Met Pro Gly Gly
225 230 235 240
Phe Thr Ile Thr Val Ala Arg Lys Tyr Leu Gln Thr Arg Trp Gly Leu
245 250 255
Pro Ser Gly Arg Gln Asp Gly Val Leu Leu Val Ala Leu Ser Asn Glu
260 265 270
Pro Ala Ala Arg Leu Gly Ser Glu Ala Asp Ala Lys Ala Phe Leu Asp
275 280 285
Ser Met Ala Gln Lys Tyr Ala Ser Ile Val Gly Val Asp Leu Ser Ser
290 295 300
Ala Ala Gly Gly Gly Ala Ala Ala Ala Ala Pro Ala Met Ala Phe Ala
305 310 315 320
Ala Pro Ala Ala Ala Gly Gly Gly Ala Pro Ala Lys Thr Ile Leu Ile
325 330 335
Leu Gly Asp Ser Leu Ser Ala Gly Tyr Gly Ile Asn Pro Glu Gln Gly
340 345 350
Trp Val Ala Leu Leu Gln Lys Arg Leu Asp Gln Gln Phe Pro Lys Gln
355 360 365
His Lys Val Ile Asn Ala Ser Val Ser Gly Glu Thr Thr Ser Gly Ala
370 375 380
Leu Ala Arg Leu Pro Lys Leu Leu Thr Thr Tyr Arg Pro Asn Val Val
385 390 395 400
Val Ile Glu Leu Gly Gly Asn Asp Ala Leu Arg Gly Gln Pro Pro Gln
405 410 415
Met Ile Gln Ser Asn Leu Glu Lys Leu Ile Gln His Ser Gln Lys Ala
420 425 430
Lys Ser Lys Val Val Val Phe Gly Met Lys Ile Pro Pro Asn Tyr Gly
435 440 445
Thr Ala Tyr Ser Gln Ala Phe Glu Asn Asn Tyr Lys Val Val Ser Gln
450 455 460
Thr Tyr Gln Val Lys Leu Leu Pro Phe Phe Leu Asp Gly Val Ala Gly
465 470 475 480
His Lys Ser Leu Met Gln Asn Asp Gln Ile His Pro Asn Ala Lys Ala
485 490 495
Gln Ser Ile Leu Leu Asn Asn Ala Tyr Pro Tyr Ile Lys Gly Ala Leu
500 505 510
Ala Gly Ala Ser Ala Ser Gly Ala Ala Gly Ala Gly Ala Ala Ala Gly
515 520 525
Ala Ala Met Ile Asp Ala Gly Ala Leu Glu Glu Ile Thr Lys Asp His
530 535 540
Lys Val Leu Ala Arg Gln Gln Leu Gln Val Leu Ala Arg Tyr Leu Lys
545 550 555 560
Met Asp Leu Asp Asn Gly Glu Arg Lys Phe Leu Lys Glu Lys Asp Thr
565 570 575
Val Ala Glu Leu Gln Ala Gln Leu Asp Tyr Leu Asn Ala Glu Leu Gly
580 585 590
Glu Phe Phe Val Asn Gly Val Ala Thr Ser Phe Ser Arg Lys Lys Ala
595 600 605
Arg Thr Phe Asp Ser Ser Trp Asn Trp Ala Lys Gln Ser Leu Leu Ser
610 615 620
Leu Tyr Phe Glu Ile Ile His Gly Val Leu Lys Asn Val Asp Arg Glu
625 630 635 640
Val Val Ser Glu Ala Ile Asn Ile Met Asn Arg Ser Asn Asp Ala Leu
645 650 655
Ile Lys Phe Met Glu Tyr His Ile Ser Asn Thr Asp Glu Thr Lys Gly
660 665 670
Glu Asn Tyr Gln Leu Val Lys Thr Leu Gly Glu Gln Leu Ile Glu Asn
675 680 685
Cys Lys Gln Val Leu Asp Val Asp Pro Val Tyr Lys Asp Val Ala Lys
690 695 700
Pro Thr Gly Pro Lys Thr Ala Ile Asp Lys Asn Gly Asn Ile Thr Tyr
705 710 715 720
Ser Glu Glu Pro Arg Glu Lys Val Arg Lys Leu Ser Gln Tyr Val Gln
725 730 735
Glu Met Ala Leu Gly Gly Pro Ile Thr Lys Glu Ser Gln Pro Thr Ile
740 745 750
Glu Glu Asp Leu Thr Arg Val Tyr Lys Ala Ile Ser Ala Gln Ala Asp
755 760 765
Lys Gln Asp Ile Ser Ser Ser Thr Arg Val Glu Phe Glu Lys Leu Tyr
770 775 780
Ser Asp Leu Met Lys Phe Leu Glu Ser Ser Lys Glu Ile Asp Pro Ser
785 790 795 800
Gln Thr Thr Gln Leu Ala Gly Met Asp Val Glu Asp Ala Leu Asp Lys
805 810 815
Asp Ser Thr Lys Glu Val Ala Ser Leu Pro Asn Lys Ser Thr Ile Ser
820 825 830
Lys Thr Val Ser Ser Thr Ile Pro Arg Glu Thr Ile Pro Phe Leu His
835 840 845
Leu Arg Lys Lys Thr Pro Ala Gly Asp Trp Lys Tyr Asp Arg Gln Leu
850 855 860
Ser Ser Leu Phe Leu Asp Gly Leu Glu Lys Ala Ala Phe Asn Gly Val
865 870 875 880
Thr Phe Lys Asp Lys Tyr Val Leu Ile Thr Gly Ala Gly Lys Gly Ser
885 890 895
Ile Gly Ala Glu Val Leu Gln Gly Leu Leu Gln Gly Gly Ala Lys Val
900 905 910
Val Val Thr Thr Ser Arg Phe Ser Lys Gln Val Thr Asp Tyr Tyr Gln
915 920 925
Ser Ile Tyr Ala Lys Tyr Gly Ala Lys Gly Ser Thr Leu Ile Val Val
930 935 940
Pro Phe Asn Gln Gly Ser Lys Gln Asp Val Glu Ala Leu Ile Glu Phe
945 950 955 960
Ile Tyr Asp Thr Glu Lys Asn Gly Gly Leu Gly Trp Asp Leu Asp Ala
965 970 975
Ile Ile Pro Phe Ala Ala Ile Pro Glu Gln Gly Ile Glu Leu Glu His
980 985 990
Ile Asp Ser Lys Ser Glu Phe Ala His Arg Ile Met Leu Thr Asn Ile
995 1000 1005
Leu Arg Met Met Gly Cys Val Lys Lys Gln Lys Ser Ala Arg Gly
1010 1015 1020
Ile Glu Thr Arg Pro Ala Gln Val Ile Leu Pro Met Ser Pro Asn
1025 1030 1035
His Gly Thr Phe Gly Gly Asp Gly Met Tyr Ser Glu Ser Lys Leu
1040 1045 1050
Ser Leu Glu Thr Leu Phe Asn Arg Trp His Ser Glu Ser Trp Ala
1055 1060 1065
Asn Gln Leu Thr Val Cys Gly Ala Ile Ile Gly Trp Thr Arg Gly
1070 1075 1080
Thr Gly Leu Met Ser Ala Asn Asn Ile Ile Ala Glu Gly Ile Glu
1085 1090 1095
Lys Met Gly Val Arg Thr Phe Ser Gln Lys Glu Met Ala Phe Asn
1100 1105 1110
Leu Leu Gly Leu Leu Thr Pro Glu Val Val Glu Leu Cys Gln Lys
1115 1120 1125
Ser Pro Val Met Ala Asp Leu Asn Gly Gly Leu Gln Phe Val Pro
1130 1135 1140
Glu Leu Lys Glu Phe Thr Ala Lys Leu Arg Lys Glu Leu Val Glu
1145 1150 1155
Thr Ser Glu Val Arg Lys Ala Val Ser Ile Glu Thr Ala Leu Glu
1160 1165 1170
His Lys Val Val Asn Gly Asn Ser Ala Asp Ala Ala Tyr Ala Gln
1175 1180 1185
Val Glu Ile Gln Pro Arg Ala Asn Ile Gln Leu Asp Phe Pro Glu
1190 1195 1200
Leu Lys Pro Tyr Lys Gln Val Lys Gln Ile Ala Pro Ala Glu Leu
1205 1210 1215
Glu Gly Leu Leu Asp Leu Glu Arg Val Ile Val Val Thr Gly Phe
1220 1225 1230
Ala Glu Val Gly Pro Trp Gly Ser Ala Arg Thr Arg Trp Glu Met
1235 1240 1245
Glu Ala Phe Gly Glu Phe Ser Leu Glu Gly Cys Val Glu Met Ala
1250 1255 1260
Trp Ile Met Gly Phe Ile Ser Tyr His Asn Gly Asn Leu Lys Gly
1265 1270 1275
Arg Pro Tyr Thr Gly Trp Val Asp Ser Lys Thr Lys Glu Pro Val
1280 1285 1290
Asp Asp Lys Asp Val Lys Ala Lys Tyr Glu Thr Ser Ile Leu Glu
1295 1300 1305
His Ser Gly Ile Arg Leu Ile Glu Pro Glu Leu Phe Asn Gly Tyr
1310 1315 1320
Asn Pro Glu Lys Lys Glu Met Ile Gln Glu Val Ile Val Glu Glu
1325 1330 1335
Asp Leu Glu Pro Phe Glu Ala Ser Lys Glu Thr Ala Glu Gln Phe
1340 1345 1350
Lys His Gln His Gly Asp Lys Val Asp Ile Phe Glu Ile Pro Glu
1355 1360 1365
Thr Gly Glu Tyr Ser Val Lys Leu Leu Lys Gly Ala Thr Leu Tyr
1370 1375 1380
Ile Pro Lys Ala Leu Arg Phe Asp Arg Leu Val Ala Gly Gln Ile
1385 1390 1395
Pro Thr Gly Trp Asn Ala Lys Thr Tyr Gly Ile Ser Asp Asp Ile
1400 1405 1410
Ile Ser Gln Val Asp Pro Ile Thr Leu Phe Val Leu Val Ser Val
1415 1420 1425
Val Glu Ala Phe Ile Ala Ser Gly Ile Thr Asp Pro Tyr Glu Met
1430 1435 1440
Tyr Lys Tyr Val His Val Ser Glu Val Gly Asn Cys Ser Gly Ser
1445 1450 1455
Gly Met Gly Gly Val Ser Ala Leu Arg Gly Met Phe Lys Asp Arg
1460 1465 1470
Phe Lys Asp Glu Pro Val Gln Asn Asp Ile Leu Gln Glu Ser Phe
1475 1480 1485
Ile Asn Thr Met Ser Ala Trp Val Asn Met Leu Leu Ile Ser Ser
1490 1495 1500
Ser Gly Pro Ile Lys Thr Pro Val Gly Ala Cys Ala Thr Ser Val
1505 1510 1515
Glu Ser Val Asp Ile Gly Val Glu Thr Ile Leu Ser Gly Lys Ala
1520 1525 1530
Arg Ile Cys Ile Val Gly Gly Tyr Asp Asp Phe Gln Glu Glu Gly
1535 1540 1545
Ser Phe Glu Phe Gly Asn Met Lys Ala Thr Ser Asn Thr Leu Glu
1550 1555 1560
Glu Phe Glu His Gly Arg Thr Pro Ala Glu Met Ser Arg Pro Ala
1565 1570 1575
Thr Thr Thr Arg Asn Gly Phe Met Glu Ala Gln Gly Ala Gly Ile
1580 1585 1590
Gln Ile Ile Met Gln Ala Asp Leu Ala Leu Lys Met Gly Val Pro
1595 1600 1605
Ile Tyr Gly Ile Val Ala Met Ala Ala Thr Ala Thr Asp Lys Ile
1610 1615 1620
Gly Arg Ser Val Pro Ala Pro Gly Lys Gly Ile Leu Thr Thr Ala
1625 1630 1635
Arg Glu His His Ser Ser Val Lys Tyr Ala Ser Pro Asn Leu Asn
1640 1645 1650
Met Lys Tyr Arg Lys Arg Gln Leu Val Thr Arg Glu Ala Gln Ile
1655 1660 1665
Lys Asp Trp Val Glu Asn Glu Leu Glu Ala Leu Lys Leu Glu Ala
1670 1675 1680
Glu Glu Ile Pro Ser Glu Asp Gln Asn Glu Phe Leu Leu Glu Arg
1685 1690 1695
Thr Arg Glu Ile His Asn Glu Ala Glu Ser Gln Leu Arg Ala Ala
1700 1705 1710
Gln Gln Gln Trp Gly Asn Asp Phe Tyr Lys Arg Asp Pro Arg Ile
1715 1720 1725
Ala Pro Leu Arg Gly Ala Leu Ala Thr Tyr Gly Leu Thr Ile Asp
1730 1735 1740
Asp Leu Gly Val Ala Ser Phe His Gly Thr Ser Thr Lys Ala Asn
1745 1750 1755
Asp Lys Asn Glu Ser Ala Thr Ile Asn Glu Met Met Lys His Leu
1760 1765 1770
Gly Arg Ser Glu Gly Asn Pro Val Ile Gly Val Phe Gln Lys Phe
1775 1780 1785
Leu Thr Gly His Pro Lys Gly Ala Ala Gly Ala Trp Met Met Asn
1790 1795 1800
Gly Ala Leu Gln Ile Leu Asn Ser Gly Ile Ile Pro Gly Asn Arg
1805 1810 1815
Asn Ala Asp Asn Val Asp Lys Ile Leu Glu Gln Phe Glu Tyr Val
1820 1825 1830
Leu Tyr Pro Ser Lys Thr Leu Lys Thr Asp Gly Val Arg Ala Val
1835 1840 1845
Ser Ile Thr Ser Phe Gly Phe Gly Gln Lys Gly Gly Gln Ala Ile
1850 1855 1860
Val Val His Pro Asp Tyr Leu Tyr Gly Ala Ile Thr Glu Asp Arg
1865 1870 1875
Tyr Asn Glu Tyr Val Ala Lys Val Ser Ala Arg Glu Lys Ser Ala
1880 1885 1890
Tyr Lys Phe Phe His Asn Gly Met Ile Tyr Asn Lys Leu Phe Val
1895 1900 1905
Ser Lys Glu His Ala Pro Tyr Thr Asp Glu Leu Glu Glu Asp Val
1910 1915 1920
Tyr Leu Asp Pro Leu Ala Arg Val Ser Lys Asp Lys Lys Ser Gly
1925 1930 1935
Ser Leu Thr Phe Asn Ser Lys Asn Ile Gln Ser Lys Asp Ser Tyr
1940 1945 1950
Ile Asn Ala Asn Thr Ile Glu Thr Ala Lys Met Ile Glu Asn Met
1955 1960 1965
Thr Lys Glu Lys Val Ser Asn Gly Gly Val Gly Val Asp Val Glu
1970 1975 1980
Leu Ile Thr Ser Ile Asn Val Glu Asn Asp Thr Phe Ile Glu Arg
1985 1990 1995
Asn Phe Thr Pro Gln Glu Ile Glu Tyr Cys Ser Ala Gln Pro Ser
2000 2005 2010
Val Gln Ser Ser Phe Ala Gly Thr Trp Ser Ala Lys Glu Ala Val
2015 2020 2025
Phe Lys Ser Leu Gly Val Lys Ser Leu Gly Gly Gly Ala Ala Leu
2030 2035 2040
Lys Asp Ile Glu Ile Val Arg Val Asn Lys Asn Ala Pro Ala Val
2045 2050 2055
Glu Leu His Gly Asn Ala Lys Lys Ala Ala Glu Glu Ala Gly Val
2060 2065 2070
Thr Asp Val Lys Val Ser Ile Ser His Asp Asp Leu Gln Ala Val
2075 2080 2085
Ala Val Ala Val Ser Thr Lys Lys
2090 2095
<210> 19
<211> 6156
<212> DNA
<213> 酿酒酵母
<400> 19
atggacgctt actccacaag accattaacc ctatctcacg gttctttaga gcacgtgctt 60
ctggtaccaa ccgcttcatt tttcattgct tcgcaattac aagaacaatt taataaaatt 120
ttgcccgaac ccactgaagg gtttgctgca gatgacgagc ctaccacacc tgctgaacta 180
gtggggaaat tccttggcta cgtatcttct ctagtcgaac cttccaaggt cggtcaattc 240
gatcaggtct tgaacctttg cttaacagaa tttgaaaact gttatttaga aggcaatgac 300
attcacgcct tggctgctaa actattacag gaaaacgaca caactttagt gaagactaaa 360
gaactaatta aaaattatat taccgccaga ataatggcta agagaccatt tgacaaaaaa 420
tccaactctg ctctttttag ggccgtcggc gagggtaacg cacaattggt agccattttc 480
ggtggtcaag gtaacaccga cgactacttt gaagaattgc gtgatctata tcaaacttat 540
catgtcttag tgggagattt aatcaagttc tccgctgaaa ctttaagtga actgattaga 600
actactttag atgctgaaaa agtctttact caaggtttaa acatattgga atggttggag 660
aacccttcaa ataccccaga caaggactat ttactttcca ttccaatttc atgcccctta 720
attggtgtca ttcaattggc tcactacgta gttactgcca agcttttggg tttcactcca 780
ggtgagttaa gatcttactt aaaaggtgct acaggtcact ctcaaggttt ggttactgct 840
gtcgccatag ctgagacgga ttcctgggaa tccttcttcg tctccgtaag aaaagcaatt 900
actgtattat tcttcatcgg tgttcgttgt tacgaagcat acccaaacac ttccctacca 960
ccatccatct tggaagattc cttggaaaac aatgaaggtg ttccatctcc aatgttgtcc 1020
atttccaatc taactcaaga acaagttcaa gactatgtaa ataagactaa ctctcatttg 1080
ccagctggta aacaagttga aatttctcta gtcaatggtg cgaagaatct agtcgtatcg 1140
ggcccaccac aatcattata tggtttaaac ttgactttaa gaaaggccaa ggccccatct 1200
ggactggatc aatcaagaat cccattcagc gaaagaaaat tgaagttctc caataggttc 1260
ttacctgttg catcaccatt ccattcccat ctattggttc cagcttcaga tttgattaac 1320
aaagacttag tcaaaaacaa tgtcagcttt aacgctaaag atattcaaat ccccgtttac 1380
gacacttttg atggttcaga tctaagagtc ctttcaggtt ccatttccga gagaatcgtc 1440
gactgcatca ttagattacc tgtcaaatgg gaaactacta cacaattcaa agccacccac 1500
atattagact ttggtccagg tggagcttcc ggtttaggtg ttttaaccca tcgtaataaa 1560
gatggtactg gtgttcgtgt tatcgttgcc ggtactctcg acattaaccc agatgatgat 1620
tacggattca agcaagaaat ctttgatgtt actagtaatg gtttgaagaa aaatccaaac 1680
tggttggaag aataccatcc aaaattaatt aagaacaaat caggcaaaat ttttgtcgaa 1740
acaaaatttt ctaaattaat cggtagacca cctttattgg ttcctggtat gacaccatgt 1800
actgtttctc cagatttcgt agctgctacc acaaatgctg gttataccat tgagttggcc 1860
ggtggtggtt acttttccgc agcaggtatg accgccgcta ttgattctgt ggtttctcag 1920
atagaaaagg gtagtacctt cggtatcaac ttgatctacg tcaatccatt tatgttacaa 1980
tggggtattc cattaatcaa ggaactaaga agcaaaggtt atccaattca attcttgacc 2040
attggtgctg gtgtcccatc attggaagtt gctagtgaat acatagagac attaggtttg 2100
aagtacttgg gtttgaaacc aggttccatt gatgctattt cgcaagttat aaacattgct 2160
aaagcacatc caaacttccc aatagcttta caatggaccg gtggtagagg tggtggtcat 2220
cattctttcg aagatgccca cactccaatg ttacaaatgt actccaagat tagaagacat 2280
ccaaacatta tgttgatatt cggttctggt ttcggttctg ctgatgacac ttacccatac 2340
ttaaccggtg aatggtccac aaaattcgat tatccaccaa tgccattcga tggtttccta 2400
tttggttcga gggtcatgat tgctaaggaa gttaaaactt ctcctgatgc taagaagtgt 2460
attgctgctt gtactggtgt tcctgatgat aaatgggaac aaacctacaa gaagccaact 2520
ggtggtattg tcactgttcg ctctgaaatg ggtgaaccaa ttcacaaaat tgccactcgt 2580
ggtgttatgc tatggaagga attcgacgaa accatcttca acttaccaaa gaataagttg 2640
gtaccaactt tggaagcaaa gagagattac attatctcaa gattgaacgc cgatttccaa 2700
aaaccatggt ttgctaccgt caacggtcaa gcccgtgacc tagccacaat gacatacgaa 2760
gaagttgcaa agagattggt ggaattaatg ttcatcagat ctaccaactc ttggtttgat 2820
gtcacatgga gaacctttac tggtgatttc ctacgtcgtg tcgaagaacg tttcactaaa 2880
agtaagacat tgtctttaat ccaatcctat tctctactag acaagcctga tgaagctatt 2940
gaaaaagtat ttaatgctta tcctgccgct agggaacagt tcttgaatgc gcaagatatt 3000
gatcactttt tgagcatgtg tcaaaatcca atgcaaaaac cagtgccttt tgttccagtt 3060
ttggatcgta gattcgagat ttttttcaaa aaagattcgt tatggcaatc tgagcacttg 3120
gaagccgtcg tcgaccaaga cgttcaaaga acatgtatcc tacatggacc tgttgcagca 3180
caattcacta aagtcatcga tgaaccaatt aagagcatta tggatggtat tcacgatggt 3240
cacatcaaaa agttactaca tcaatattac ggtgacgatg agtcaaagat tccagcagtt 3300
gagtactttg gtggtgaaag ccctgtagac gtacaaagtc aagttgattc ttcctctgta 3360
tctgaagact cagctgtttt taaggcaaca tcctctactg atgaagaaag ctggtttaag 3420
gctttggcgg gatccgaaat taactggaga catgcaagtt tcttatgttc ctttatcact 3480
caagataaaa tgtttgtttc taacccaatt agaaaagttt tcaagccaag ccaaggaatg 3540
gttgttgaga tttccaacgg caatacttct tcaaagactg ttgtcactct ttcagaacct 3600
gttcaaggtg aattgaaacc aactgttatt ttgaagttgt tgaaggagaa cataatccaa 3660
atggaaatga ttgagaacag aactatggat ggtaagcccg tcagcttgcc attgttgtac 3720
aacttcaacc cagataatgg ttttgctcca atctctgaag ttatggagga cagaaaccaa 3780
agaattaagg aaatgtactg gaaattatgg attgatgagc ctttcaattt ggactttgac 3840
ccaagagatg tcattaaggg caaagatttc gagatcaccg ctaaagaagt ttatgacttt 3900
acacacgctg ttggaaacaa ttgtgaagac ttcgtttcta gacctgatag aacgatgttg 3960
gccccaatgg actttgctat tgttgtcgga tggagagcca tcatcaaggc cattttccct 4020
aatacggtcg atggtgactt attgaagttg gttcatttgt ctaacggcta caagatgatt 4080
cctggcgcta agccactgca agttggtgat gttgtttcaa ctactgctgt tattgaatct 4140
gtcgtcaacc aacctacagg aaagattgtc gatgtggtag gtacattatc gagaaatggc 4200
aagcctgtca tggaagtcac ctcctcattc ttctacagag gcaactatac tgactttgaa 4260
aacactttcc aaaagactgt tgaacctgtt tatcaaatgc acatcaaaac ttctaaagat 4320
atagctgtct tgcgctctaa ggagtggttc caattggacg atgaagactt cgatctgtta 4380
aacaaaactt tgactttcga aactgaaact gaagttactt tcaagaatgc taacatcttc 4440
tcttcagtga aatgttttgg cccaattaaa gttgaattgc caaccaaaga aaccgtggag 4500
atcggtattg tcgattacga agccggtgcc tctcacggta accctgttgt tgatttcttg 4560
aagagaaacg gttccacatt ggaacaaaag gtcaatctag aaaatcctat tccaattgca 4620
gtacttgatt cgtacactcc aagtaccaac gaaccatacg ctagagtttc tggtgatttg 4680
aatccaattc acgtttcacg tcattttgcc tcttacgcaa acttgccagg tactatcacg 4740
cacggtatgt tttcttctgc ttccgtccgt gctttgattg aaaactgggc tgctgacagt 4800
gtttcatcca gggtacgtgg ctacacttgt caatttgttg acatggtttt gcctaacact 4860
gctttgaaaa catcgattca acatgttggt atgatcaatg gtagaaaatt gataaagttt 4920
gaaactagaa atgaagatga cgttgtagtt ttgactggtg aagccgaaat tgaacaacct 4980
gttactacct tcgttttcac tggtcaaggt tcacaagaac aaggtatggg tatggactta 5040
tacaaaactt ctaaagctgc tcaagatgtt tggaatagag ctgacaacca tttcaaggac 5100
acttatggtt tctctatctt agacattgtc attaacaacc cagttaactt aacaattcac 5160
ttcggtggtg aaaagggtaa gaggatcaga gaaaactatt ctgctatgat ctttgagact 5220
atcgtggatg gaaaattgaa gactgaaaaa attttcaagg aaattaatga gcacagtact 5280
tcttacacat ttagatctga aaaaggttta ttgtctgcta ctcaatttac acaaccagct 5340
ttaactttga tggaaaaagc tgctttcgaa gacttgaaat ctaaaggttt gatcccagcc 5400
gatgctactt ttgctggtca ctctttaggt gagtatgctg ctttggcctc tttggctgat 5460
gttatgtcta tcgaatcttt agttgaagtt gtgttctaca gaggtatgac tatgcaagtt 5520
gctgttccaa gagatgagtt gggcagatcc aactatggta tgattgccat taacccaggt 5580
agagtcgctg catcattctc tcaagaagct ttgcaatatg ttgttgagag agttggtaag 5640
agaaccggct ggttggttga aatcgtcaac tacaacgttg aaaaccaaca atatgttgca 5700
gctggtgatc taagagcttt agacaccgtt accaatgttc taaacttcat caaattacaa 5760
aaaattgata ttattgaact acaaaagtcc ttatctttgg aagaagttga aggtcatttg 5820
tttgagatca ttgacgaagc ttccaagaaa tctgctgtca agcctcgccc acttaaattg 5880
gagagaggtt ttgcttgtat cccattagtt ggtatttctg ttcctttcca ttccacctac 5940
ttgatgaatg gtgttaaacc attcaagagt ttcttgaaga agaatatcat aaaagaaaat 6000
gtgaaggttg ctagattggc cggaaagtac attccaaact tgactgcaaa accattccag 6060
gttactaagg aatatttcca ggacgtttat gatttgactg gctccgaacc tatcaaggaa 6120
atcatcgaca actgggaaaa gtatgaacaa tcctaa 6156
<210> 20
<211> 2051
<212> PRT
<213> 酿酒酵母
<400> 20
Met Asp Ala Tyr Ser Thr Arg Pro Leu Thr Leu Ser His Gly Ser Leu
1 5 10 15
Glu His Val Leu Leu Val Pro Thr Ala Ser Phe Phe Ile Ala Ser Gln
20 25 30
Leu Gln Glu Gln Phe Asn Lys Ile Leu Pro Glu Pro Thr Glu Gly Phe
35 40 45
Ala Ala Asp Asp Glu Pro Thr Thr Pro Ala Glu Leu Val Gly Lys Phe
50 55 60
Leu Gly Tyr Val Ser Ser Leu Val Glu Pro Ser Lys Val Gly Gln Phe
65 70 75 80
Asp Gln Val Leu Asn Leu Cys Leu Thr Glu Phe Glu Asn Cys Tyr Leu
85 90 95
Glu Gly Asn Asp Ile His Ala Leu Ala Ala Lys Leu Leu Gln Glu Asn
100 105 110
Asp Thr Thr Leu Val Lys Thr Lys Glu Leu Ile Lys Asn Tyr Ile Thr
115 120 125
Ala Arg Ile Met Ala Lys Arg Pro Phe Asp Lys Lys Ser Asn Ser Ala
130 135 140
Leu Phe Arg Ala Val Gly Glu Gly Asn Ala Gln Leu Val Ala Ile Phe
145 150 155 160
Gly Gly Gln Gly Asn Thr Asp Asp Tyr Phe Glu Glu Leu Arg Asp Leu
165 170 175
Tyr Gln Thr Tyr His Val Leu Val Gly Asp Leu Ile Lys Phe Ser Ala
180 185 190
Glu Thr Leu Ser Glu Leu Ile Arg Thr Thr Leu Asp Ala Glu Lys Val
195 200 205
Phe Thr Gln Gly Leu Asn Ile Leu Glu Trp Leu Glu Asn Pro Ser Asn
210 215 220
Thr Pro Asp Lys Asp Tyr Leu Leu Ser Ile Pro Ile Ser Cys Pro Leu
225 230 235 240
Ile Gly Val Ile Gln Leu Ala His Tyr Val Val Thr Ala Lys Leu Leu
245 250 255
Gly Phe Thr Pro Gly Glu Leu Arg Ser Tyr Leu Lys Gly Ala Thr Gly
260 265 270
His Ser Gln Gly Leu Val Thr Ala Val Ala Ile Ala Glu Thr Asp Ser
275 280 285
Trp Glu Ser Phe Phe Val Ser Val Arg Lys Ala Ile Thr Val Leu Phe
290 295 300
Phe Ile Gly Val Arg Cys Tyr Glu Ala Tyr Pro Asn Thr Ser Leu Pro
305 310 315 320
Pro Ser Ile Leu Glu Asp Ser Leu Glu Asn Asn Glu Gly Val Pro Ser
325 330 335
Pro Met Leu Ser Ile Ser Asn Leu Thr Gln Glu Gln Val Gln Asp Tyr
340 345 350
Val Asn Lys Thr Asn Ser His Leu Pro Ala Gly Lys Gln Val Glu Ile
355 360 365
Ser Leu Val Asn Gly Ala Lys Asn Leu Val Val Ser Gly Pro Pro Gln
370 375 380
Ser Leu Tyr Gly Leu Asn Leu Thr Leu Arg Lys Ala Lys Ala Pro Ser
385 390 395 400
Gly Leu Asp Gln Ser Arg Ile Pro Phe Ser Glu Arg Lys Leu Lys Phe
405 410 415
Ser Asn Arg Phe Leu Pro Val Ala Ser Pro Phe His Ser His Leu Leu
420 425 430
Val Pro Ala Ser Asp Leu Ile Asn Lys Asp Leu Val Lys Asn Asn Val
435 440 445
Ser Phe Asn Ala Lys Asp Ile Gln Ile Pro Val Tyr Asp Thr Phe Asp
450 455 460
Gly Ser Asp Leu Arg Val Leu Ser Gly Ser Ile Ser Glu Arg Ile Val
465 470 475 480
Asp Cys Ile Ile Arg Leu Pro Val Lys Trp Glu Thr Thr Thr Gln Phe
485 490 495
Lys Ala Thr His Ile Leu Asp Phe Gly Pro Gly Gly Ala Ser Gly Leu
500 505 510
Gly Val Leu Thr His Arg Asn Lys Asp Gly Thr Gly Val Arg Val Ile
515 520 525
Val Ala Gly Thr Leu Asp Ile Asn Pro Asp Asp Asp Tyr Gly Phe Lys
530 535 540
Gln Glu Ile Phe Asp Val Thr Ser Asn Gly Leu Lys Lys Asn Pro Asn
545 550 555 560
Trp Leu Glu Glu Tyr His Pro Lys Leu Ile Lys Asn Lys Ser Gly Lys
565 570 575
Ile Phe Val Glu Thr Lys Phe Ser Lys Leu Ile Gly Arg Pro Pro Leu
580 585 590
Leu Val Pro Gly Met Thr Pro Cys Thr Val Ser Pro Asp Phe Val Ala
595 600 605
Ala Thr Thr Asn Ala Gly Tyr Thr Ile Glu Leu Ala Gly Gly Gly Tyr
610 615 620
Phe Ser Ala Ala Gly Met Thr Ala Ala Ile Asp Ser Val Val Ser Gln
625 630 635 640
Ile Glu Lys Gly Ser Thr Phe Gly Ile Asn Leu Ile Tyr Val Asn Pro
645 650 655
Phe Met Leu Gln Trp Gly Ile Pro Leu Ile Lys Glu Leu Arg Ser Lys
660 665 670
Gly Tyr Pro Ile Gln Phe Leu Thr Ile Gly Ala Gly Val Pro Ser Leu
675 680 685
Glu Val Ala Ser Glu Tyr Ile Glu Thr Leu Gly Leu Lys Tyr Leu Gly
690 695 700
Leu Lys Pro Gly Ser Ile Asp Ala Ile Ser Gln Val Ile Asn Ile Ala
705 710 715 720
Lys Ala His Pro Asn Phe Pro Ile Ala Leu Gln Trp Thr Gly Gly Arg
725 730 735
Gly Gly Gly His His Ser Phe Glu Asp Ala His Thr Pro Met Leu Gln
740 745 750
Met Tyr Ser Lys Ile Arg Arg His Pro Asn Ile Met Leu Ile Phe Gly
755 760 765
Ser Gly Phe Gly Ser Ala Asp Asp Thr Tyr Pro Tyr Leu Thr Gly Glu
770 775 780
Trp Ser Thr Lys Phe Asp Tyr Pro Pro Met Pro Phe Asp Gly Phe Leu
785 790 795 800
Phe Gly Ser Arg Val Met Ile Ala Lys Glu Val Lys Thr Ser Pro Asp
805 810 815
Ala Lys Lys Cys Ile Ala Ala Cys Thr Gly Val Pro Asp Asp Lys Trp
820 825 830
Glu Gln Thr Tyr Lys Lys Pro Thr Gly Gly Ile Val Thr Val Arg Ser
835 840 845
Glu Met Gly Glu Pro Ile His Lys Ile Ala Thr Arg Gly Val Met Leu
850 855 860
Trp Lys Glu Phe Asp Glu Thr Ile Phe Asn Leu Pro Lys Asn Lys Leu
865 870 875 880
Val Pro Thr Leu Glu Ala Lys Arg Asp Tyr Ile Ile Ser Arg Leu Asn
885 890 895
Ala Asp Phe Gln Lys Pro Trp Phe Ala Thr Val Asn Gly Gln Ala Arg
900 905 910
Asp Leu Ala Thr Met Thr Tyr Glu Glu Val Ala Lys Arg Leu Val Glu
915 920 925
Leu Met Phe Ile Arg Ser Thr Asn Ser Trp Phe Asp Val Thr Trp Arg
930 935 940
Thr Phe Thr Gly Asp Phe Leu Arg Arg Val Glu Glu Arg Phe Thr Lys
945 950 955 960
Ser Lys Thr Leu Ser Leu Ile Gln Ser Tyr Ser Leu Leu Asp Lys Pro
965 970 975
Asp Glu Ala Ile Glu Lys Val Phe Asn Ala Tyr Pro Ala Ala Arg Glu
980 985 990
Gln Phe Leu Asn Ala Gln Asp Ile Asp His Phe Leu Ser Met Cys Gln
995 1000 1005
Asn Pro Met Gln Lys Pro Val Pro Phe Val Pro Val Leu Asp Arg
1010 1015 1020
Arg Phe Glu Ile Phe Phe Lys Lys Asp Ser Leu Trp Gln Ser Glu
1025 1030 1035
His Leu Glu Ala Val Val Asp Gln Asp Val Gln Arg Thr Cys Ile
1040 1045 1050
Leu His Gly Pro Val Ala Ala Gln Phe Thr Lys Val Ile Asp Glu
1055 1060 1065
Pro Ile Lys Ser Ile Met Asp Gly Ile His Asp Gly His Ile Lys
1070 1075 1080
Lys Leu Leu His Gln Tyr Tyr Gly Asp Asp Glu Ser Lys Ile Pro
1085 1090 1095
Ala Val Glu Tyr Phe Gly Gly Glu Ser Pro Val Asp Val Gln Ser
1100 1105 1110
Gln Val Asp Ser Ser Ser Val Ser Glu Asp Ser Ala Val Phe Lys
1115 1120 1125
Ala Thr Ser Ser Thr Asp Glu Glu Ser Trp Phe Lys Ala Leu Ala
1130 1135 1140
Gly Ser Glu Ile Asn Trp Arg His Ala Ser Phe Leu Cys Ser Phe
1145 1150 1155
Ile Thr Gln Asp Lys Met Phe Val Ser Asn Pro Ile Arg Lys Val
1160 1165 1170
Phe Lys Pro Ser Gln Gly Met Val Val Glu Ile Ser Asn Gly Asn
1175 1180 1185
Thr Ser Ser Lys Thr Val Val Thr Leu Ser Glu Pro Val Gln Gly
1190 1195 1200
Glu Leu Lys Pro Thr Val Ile Leu Lys Leu Leu Lys Glu Asn Ile
1205 1210 1215
Ile Gln Met Glu Met Ile Glu Asn Arg Thr Met Asp Gly Lys Pro
1220 1225 1230
Val Ser Leu Pro Leu Leu Tyr Asn Phe Asn Pro Asp Asn Gly Phe
1235 1240 1245
Ala Pro Ile Ser Glu Val Met Glu Asp Arg Asn Gln Arg Ile Lys
1250 1255 1260
Glu Met Tyr Trp Lys Leu Trp Ile Asp Glu Pro Phe Asn Leu Asp
1265 1270 1275
Phe Asp Pro Arg Asp Val Ile Lys Gly Lys Asp Phe Glu Ile Thr
1280 1285 1290
Ala Lys Glu Val Tyr Asp Phe Thr His Ala Val Gly Asn Asn Cys
1295 1300 1305
Glu Asp Phe Val Ser Arg Pro Asp Arg Thr Met Leu Ala Pro Met
1310 1315 1320
Asp Phe Ala Ile Val Val Gly Trp Arg Ala Ile Ile Lys Ala Ile
1325 1330 1335
Phe Pro Asn Thr Val Asp Gly Asp Leu Leu Lys Leu Val His Leu
1340 1345 1350
Ser Asn Gly Tyr Lys Met Ile Pro Gly Ala Lys Pro Leu Gln Val
1355 1360 1365
Gly Asp Val Val Ser Thr Thr Ala Val Ile Glu Ser Val Val Asn
1370 1375 1380
Gln Pro Thr Gly Lys Ile Val Asp Val Val Gly Thr Leu Ser Arg
1385 1390 1395
Asn Gly Lys Pro Val Met Glu Val Thr Ser Ser Phe Phe Tyr Arg
1400 1405 1410
Gly Asn Tyr Thr Asp Phe Glu Asn Thr Phe Gln Lys Thr Val Glu
1415 1420 1425
Pro Val Tyr Gln Met His Ile Lys Thr Ser Lys Asp Ile Ala Val
1430 1435 1440
Leu Arg Ser Lys Glu Trp Phe Gln Leu Asp Asp Glu Asp Phe Asp
1445 1450 1455
Leu Leu Asn Lys Thr Leu Thr Phe Glu Thr Glu Thr Glu Val Thr
1460 1465 1470
Phe Lys Asn Ala Asn Ile Phe Ser Ser Val Lys Cys Phe Gly Pro
1475 1480 1485
Ile Lys Val Glu Leu Pro Thr Lys Glu Thr Val Glu Ile Gly Ile
1490 1495 1500
Val Asp Tyr Glu Ala Gly Ala Ser His Gly Asn Pro Val Val Asp
1505 1510 1515
Phe Leu Lys Arg Asn Gly Ser Thr Leu Glu Gln Lys Val Asn Leu
1520 1525 1530
Glu Asn Pro Ile Pro Ile Ala Val Leu Asp Ser Tyr Thr Pro Ser
1535 1540 1545
Thr Asn Glu Pro Tyr Ala Arg Val Ser Gly Asp Leu Asn Pro Ile
1550 1555 1560
His Val Ser Arg His Phe Ala Ser Tyr Ala Asn Leu Pro Gly Thr
1565 1570 1575
Ile Thr His Gly Met Phe Ser Ser Ala Ser Val Arg Ala Leu Ile
1580 1585 1590
Glu Asn Trp Ala Ala Asp Ser Val Ser Ser Arg Val Arg Gly Tyr
1595 1600 1605
Thr Cys Gln Phe Val Asp Met Val Leu Pro Asn Thr Ala Leu Lys
1610 1615 1620
Thr Ser Ile Gln His Val Gly Met Ile Asn Gly Arg Lys Leu Ile
1625 1630 1635
Lys Phe Glu Thr Arg Asn Glu Asp Asp Val Val Val Leu Thr Gly
1640 1645 1650
Glu Ala Glu Ile Glu Gln Pro Val Thr Thr Phe Val Phe Thr Gly
1655 1660 1665
Gln Gly Ser Gln Glu Gln Gly Met Gly Met Asp Leu Tyr Lys Thr
1670 1675 1680
Ser Lys Ala Ala Gln Asp Val Trp Asn Arg Ala Asp Asn His Phe
1685 1690 1695
Lys Asp Thr Tyr Gly Phe Ser Ile Leu Asp Ile Val Ile Asn Asn
1700 1705 1710
Pro Val Asn Leu Thr Ile His Phe Gly Gly Glu Lys Gly Lys Arg
1715 1720 1725
Ile Arg Glu Asn Tyr Ser Ala Met Ile Phe Glu Thr Ile Val Asp
1730 1735 1740
Gly Lys Leu Lys Thr Glu Lys Ile Phe Lys Glu Ile Asn Glu His
1745 1750 1755
Ser Thr Ser Tyr Thr Phe Arg Ser Glu Lys Gly Leu Leu Ser Ala
1760 1765 1770
Thr Gln Phe Thr Gln Pro Ala Leu Thr Leu Met Glu Lys Ala Ala
1775 1780 1785
Phe Glu Asp Leu Lys Ser Lys Gly Leu Ile Pro Ala Asp Ala Thr
1790 1795 1800
Phe Ala Gly His Ser Leu Gly Glu Tyr Ala Ala Leu Ala Ser Leu
1805 1810 1815
Ala Asp Val Met Ser Ile Glu Ser Leu Val Glu Val Val Phe Tyr
1820 1825 1830
Arg Gly Met Thr Met Gln Val Ala Val Pro Arg Asp Glu Leu Gly
1835 1840 1845
Arg Ser Asn Tyr Gly Met Ile Ala Ile Asn Pro Gly Arg Val Ala
1850 1855 1860
Ala Ser Phe Ser Gln Glu Ala Leu Gln Tyr Val Val Glu Arg Val
1865 1870 1875
Gly Lys Arg Thr Gly Trp Leu Val Glu Ile Val Asn Tyr Asn Val
1880 1885 1890
Glu Asn Gln Gln Tyr Val Ala Ala Gly Asp Leu Arg Ala Leu Asp
1895 1900 1905
Thr Val Thr Asn Val Leu Asn Phe Ile Lys Leu Gln Lys Ile Asp
1910 1915 1920
Ile Ile Glu Leu Gln Lys Ser Leu Ser Leu Glu Glu Val Glu Gly
1925 1930 1935
His Leu Phe Glu Ile Ile Asp Glu Ala Ser Lys Lys Ser Ala Val
1940 1945 1950
Lys Pro Arg Pro Leu Lys Leu Glu Arg Gly Phe Ala Cys Ile Pro
1955 1960 1965
Leu Val Gly Ile Ser Val Pro Phe His Ser Thr Tyr Leu Met Asn
1970 1975 1980
Gly Val Lys Pro Phe Lys Ser Phe Leu Lys Lys Asn Ile Ile Lys
1985 1990 1995
Glu Asn Val Lys Val Ala Arg Leu Ala Gly Lys Tyr Ile Pro Asn
2000 2005 2010
Leu Thr Ala Lys Pro Phe Gln Val Thr Lys Glu Tyr Phe Gln Asp
2015 2020 2025
Val Tyr Asp Leu Thr Gly Ser Glu Pro Ile Lys Glu Ile Ile Asp
2030 2035 2040
Asn Trp Glu Lys Tyr Glu Gln Ser
2045 2050
<210> 21
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> pFAS-TPI1p-F
<400> 21
gaattgggga tctacgtatg gtc 23
<210> 22
<211> 60
<212> DNA
<213> 人工序列
<220>
<223> RtFAS1-TPI1p-R
<400> 22
cgacgtccca gtcacgctcc gcgtcgctcg gccgttcatt tttagtttat gtatgtgttt 60
<210> 23
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> RtFAS1-F
<400> 23
atgaacggcc gagcgacgcg gag 23
<210> 24
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> RtFAS1-R1
<400> 24
agacggacgc aaagtggttg aagg 24
<210> 25
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> RtFAS1-R
<400> 25
tcagagcccg ccgaagacgt cgagc 25
<210> 26
<211> 27
<212> DNA
<213> 人工序列
<220>
<223> RtFAS1-F1
<400> 26
atgtctactc gatcaacggt gtcctcc 27
<210> 27
<211> 48
<212> DNA
<213> 人工序列
<220>
<223> RtFAS1-FBA1t-F
<400> 27
ctcaagctcg acgtcttcgg cgggctctga gttaattcaa attaattg 48
<210> 28
<211> 54
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ADH1t-R
<400> 28
gtcgcttgcg ccgtcgtcat cgcccagaag taggcgaatt tcttatgatt tatg 54
<210> 29
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-R
<400> 29
ctacttctgg gcgatgacga cgg 23
<210> 30
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-F1
<400> 30
caacgccgaa gtcgccaaca agc 23
<210> 31
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-F
<400> 31
atggtcgcgg cgcaggactt g 21
<210> 32
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPI2AcTesA-F1
<400> 32
gcggtggagc tgctgctgct gcc 23
<210> 33
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPII2AcTesA-R1
<400> 33
cgggcgctcc accgccagct gcg 23
<210> 34
<211> 26
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-4594-R
<400> 34
cgacaagatg caggagaagc aggaca 26
<210> 35
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-4264-F
<400> 35
gacaaggcag cgagcggtga tg 22
<210> 36
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-6628-R
<400> 36
ggtcgtcaac tacaacgtcg aggga 25
<210> 37
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-6583-F
<400> 37
gggattggag ggcgacgagg tga 23
<210> 38
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPI2AcTesA-R1
<400> 38
ggcggcaggc gcggcagcga cagga 25
<210> 39
<211> 54
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPI2AcTesA-F
<400> 39
cccgctcctg tcgctgccgc gcctgccgcc aagactatat tgatattggg tgac 54
<210> 40
<211> 61
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPI2AcTesA-R
<400> 40
tcgcgggggc agcagcagca gctccaccgc cggctaatgc acccttgatg taagggtaag 60
c 61
<210> 41
<211> 56
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPII2AcTesA-F
<400> 41
ctccggccgc agctggcggt ggagcgcccg ctaagactat attgatattg ggtgac 56
<210> 42
<211> 57
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPII2AcTesA-R
<400> 42
gacagccgcg ccgcctccgc ctccaccagc taatgcaccc ttgatgtaag ggtaagc 57
<210> 43
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-ACPII2AcTesA-F1
<400> 43
gctggtggag gcggaggcgg c 21
<210> 44
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-R1
<400> 44
atgccctggt ctgccttgtt gcc 23
<210> 45
<211> 54
<212> DNA
<213> 人工序列
<220>
<223> RtFAS2-TEF1p-R
<400> 45
gcgcgagcgg caagtcctgc gccgcgacca ttttgtaatt aaaacttaga ttag 54
<210> 46
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> TEF1p-F
<400> 46
atagcttcaa aatgtttcta ctc 23
<210> 47
<211> 61
<212> DNA
<213> 人工序列
<220>
<223> TEF1p-HA-F
<400> 47
gagtaaaaaa ggagtagaaa cattttgaag ctatggatcc ctagctccaa ttcgccctat 60
a 61
<210> 48
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> pFAS-pYX212t-R
<400> 48
gccgtaaacc actaaatcgg aacc 24
<210> 49
<211> 61
<212> DNA
<213> 人工序列
<220>
<223> TPI1p-ScFAS2-R
<400> 49
gcaaaatatg agctaattct tgctcaactt ccggcttcat ttttagttta tgtatgtgtt 60
t 61
<210> 50
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-F
<400> 50
aagccggaag ttgagcaaga attag 25
<210> 51
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-R1
<400> 51
acaggttcat cggcaatctc a 21
<210> 52
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-F1
<400> 52
tcagagccgt gtccatcact t 21
<210> 53
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-R
<400> 53
ctatttctta gtagaaacgg cgacc 25
<210> 54
<211> 48
<212> DNA
<213> 人工序列
<220>
<223> FBA1t-ScFAS2-F
<400> 54
gtcgcggtcg ccgtttctac taagaaatag gttaattcaa attaattg 48
<210> 55
<211> 52
<212> DNA
<213> 人工序列
<220>
<223> ADH1t-ScFAS1-R
<400> 55
cgacaactgg gaaaagtatg aacaatccta agcgaatttc ttatgattta tg 52
<210> 56
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> ScFAS1-R
<400> 56
ggattgttca tacttttccc ag 22
<210> 57
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> ScFAS1-F1
<400> 57
ctgctgtcaa gcctcgccca ct 22
<210> 58
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> ScFAS1-R1
<400> 58
tgtggtaggc tcgtcatctg c 21
<210> 59
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> ScFAS1-F
<400> 59
gacgcttact ccacaagacc a 21
<210> 60
<211> 54
<212> DNA
<213> 人工序列
<220>
<223> TEF1p-ScFAS1-R
<400> 60
gggttaatgg tcttgtggag taagcgtcca ttttgtaatt aaaacttaga ttag 54
<210> 61
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-ACPI2TE-R1
<400> 61
gagtcaccca atatcaatat agtcttaatc tcagcagctg ctgcagctgg 50
<210> 62
<211> 19
<212> DNA
<213> 人工序列
<220>
<223> AcTesA-F
<400> 62
aagactatat tgatattgg 19
<210> 63
<211> 46
<212> DNA
<213> 人工序列
<220>
<223> AcTesA_ScACP_F
<400> 63
ccgcagctgg cggtggagcg cccgctgccg atgaacctgt caaggc 46
<210> 64
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-R2
<400> 64
aagattgttt agcccagttc c 21
<210> 65
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-F2
<400> 65
cggtgttgct acttctttct c 21
<210> 66
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> ADH1t-F
<400> 66
gcgaatttct tatgatttat ga 22
<210> 67
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> pFAS-FBA1t-F
<400> 67
atagtttttt aatgagtatt gaatc 25
<210> 68
<211> 48
<212> DNA
<213> 人工序列
<220>
<223> AcTesA_ScACP-R
<400> 68
ggcagcagca gcagctccac cgccggcagc tgatgataag tcaacacc 48
<210> 69
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> AcTesA-R
<400> 69
accagctaat gcacccttga tg 22
<210> 70
<211> 51
<212> DNA
<213> 人工序列
<220>
<223> ScFAS2-ACPII2TE-F1
<400> 70
ccttacatca agggtgcatt agctggtgct agcgctagtg gtgctgccgg t 51
<210> 71
<211> 83
<212> DNA
<213> 人工序列
<220>
<223> pZWM-2?F
<400> 71
aagggccatg accacctgat gcaccaatta ggtaggtctg gctatgtcta tacctctggc 60
cgtcgcatcc ccggttcatt ttc 83
<210> 72
<211> 81
<212> DNA
<213> 人工序列
<220>
<223> pZWM-AmpR-R
<400> 72
ttggcaattt tttgctcttc tatataacag ttgaaatttg aataagaaca tcttctcaaa 60
gagaggcggt ttgcgtattg g 81
<210> 73
<211> 82
<212> DNA
<213> 人工序列
<220>
<223> pZWM-KlURA3-F
<400> 73
tttgagaaga tgttcttatt caaatttcaa ctgttatata gaagagcaaa aaattgccaa 60
ttcgatgatg tagtttctgg tt 82
<210> 74
<211> 81
<212> DNA
<213> 人工序列
<220>
<223> pZWM-KlURA3-R
<400> 74
cacctttcga gaggacgatg cccgtgtcta aatgattcga ccagcctaag aatgttcaac 60
gtgattctgg gtagaagatc g 81
<210> 75
<211> 84
<212> DNA
<213> 人工序列
<220>
<223> pZWM-TEF1p-F
<400> 75
gttgaacatt cttaggctgg tcgaatcatt tagacacggg catcgtcctc tcgaaaggtg 60
atagcttcaa aatgtttcta ctcc 84
<210> 76
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> TEF1p-R
<400> 76
attaaaactt agattagatt gc 22
<210> 77
<211> 62
<212> DNA
<213> 人工序列
<220>
<223> TEF1p-AkFAS-F
<400> 77
gaaagcatag caatctaatc taagttttaa ttacaaaatg gaacaacata aaactgagaa 60
ac 62
<210> 78
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-499-R
<400> 78
ttctgtcgtt gtcaggcgat gta 23
<210> 79
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-149-F
<400> 79
tgtatgaaac tggcacagat gag 23
<210> 80
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-5039-R
<400> 80
gtaaatgacg tggtattact atgga 25
<210> 81
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-4919-F
<400> 81
agttgactgt tcgtttcggt gga 23
<210> 82
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-10868-R
<400> 82
atcggtagca gtgtttgaca gagca 25
<210> 83
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-10610-F
<400> 83
tagcgggcgg gtatgatgac ttt 23
<210> 84
<211> 60
<212> DNA
<213> 人工序列
<220>
<223> ADH1t-AkFAS-R
<400> 84
ttaataataa aaatcataaa tcataagaaa ttcgcttaat ttgttattgc ttgggctact 60
<210> 85
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> ADH1t-F
<400> 85
gcgaatttct tatgatttat ga 22
<210> 86
<211> 83
<212> DNA
<213> 人工序列
<220>
<223> pZWM-ADH1t-R
<400> 86
gccagaggta tagacatagc cagacctacc taattggtgc atcaggtggt catggccctt 60
ccggtagagg tgtggtcaat aag 83
<210> 87
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> pZWM-R
<400> 87
gccagaggta tagacatagc ca 22
<210> 88
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> pZWM-F
<400> 88
gttgaacatt cttaggctgg 20
<210> 89
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-6900-R
<400> 89
aatttgtaca aagttgtttt ga 22
<210> 90
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-6800-F
<400> 90
aaaagtccaa attcgaacga a 21
<210> 91
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-6300-R
<400> 91
cttttgaagt tcattcccca c 21
<210> 92
<211> 52
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-ACPI2TE-F
<400> 92
ggagaaagtg gggaatgaac ttcaaaagaa gactatattg atattgggtg ac 52
<210> 93
<211> 54
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-ACPI2TE-R
<400> 93
cacactttcg ttcgaatttg gactttttaa tgcacccttg atgtaagggt aagc 54
<210> 94
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-8000-R
<400> 94
agcactatca actgtcaccc t 21
<210> 95
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-7900-F
<400> 95
tgtaatcgtg ccactccaga ac 22
<210> 96
<211> 52
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-ACPII2TE-F
<400> 96
gactgttcaa aacaactttg tacaaattaa gactatattg atattgggtg ac 52
<210> 97
<211> 56
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-ACPII2TE-R
<400> 97
ggactactat cacctgaaga gagagtacgt aatgcaccct tgatgtaagg gtaagc 56
<210> 98
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> AkFAS-7400-F
<400> 98
cgtactctct cttcaggtga tag 23
<210> 99
<211> 28
<212> DNA
<213> 人工序列
<220>
<223> dURA3-Up-F
<400> 99
aaacgacgtt gaaattgagg ctactgcg 28
<210> 100
<211> 60
<212> DNA
<213> 人工序列
<220>
<223> dURA3-UP-R
<400> 100
gaagaagaat gaccatacgt agatccccaa ttcggactag gatgagtagc agcacgttcc 60
<210> 101
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> TPI1p-R
<400> 101
ttagtttatg tatgtgtttt ttgta 25
<210> 102
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> pFAS-FBA1t-R
<400> 102
gtaagctact atgaaagact ttaca 25
<210> 103
<211> 56
<212> DNA
<213> 人工序列
<220>
<223> TEF1p-KanMX-F
<400> 103
gagtaaaaaa ggagtagaaa cattttgaag ctataagctt cgtacgctgc aggtcg 56
<210> 104
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> dURA3-KanMX-R
<400> 104
ctggccgcat cttctcaaat atgcttcccc gactcactat agggagaccg 50
<210> 105
<211> 26
<212> DNA
<213> 人工序列
<220>
<223> URA3-Down-F
<400> 105
gggaagcata tttgagaaga tgcggc 26
<210> 106
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> URA3-Down-R
<400> 106
ggaaacgctg ccctacacgt tcgc 24
<210> 107
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> ID-dURA3-UP
<400> 107
agggaagaca agcaacgaaa 20
<210> 108
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> ID-dURA3-DOWN
<400> 108
cgtcaaggtc tgttgagtgc 20
Claims (15)
1.重组原核或真核细胞，所述细胞包含、表达或能够表达真菌脂肪酸合酶(FAS)和硫酯酶(TE)，任选地其中所述FAS包含FAS1和FAS2。
2.根据权利要求1的重组原核或真核细胞，其中所述TE和所述FAS、FAS1或FAS2作为单一开放阅读框(ORF)编码。
3.根据权利要求1或2的重组原核或真核细胞，其中编码所述TE的多核酸序列与编码酰基载体蛋白(ACP)的多核酸序列相邻。
4.根据权利要求1至3中任一项的重组原核或真核细胞，其包含FAS编码序列或包含源自具有至少两个ACP编码核苷酸序列的FAS基因的FAS1和FAS2编码序列，其中用TE编码核苷酸序列替换所述ACP编码核苷酸序列之一。
5.根据权利要求1至4中任一项的重组原核或真核细胞，其中所述FAS来自酿酒酵母(Saccharomyces cerevisiae)、圆红冬孢酵母(Rhodosporidium toruloides)或Aplanochytrium kerguelense。
6.根据权利要求1至5中任一项的重组原核或真核细胞，其中所述TE为酰基-CoA/ACPTE，优选短或中链酰基-CoA/ACP TE。
7.根据权利要求1至6中任一项的重组原核或真核细胞，其为细菌细胞、真菌细胞或藻类细胞。
8.重组多核酸，所述多核酸编码真菌脂肪酸合酶(FAS)或它的FAS1或FAS2亚基，以及硫酯酶(TE)。
9.由根据权利要求8的重组多核酸编码的重组多肽。
10.包含根据权利要求8的重组多核酸或编码根据权利要求9的重组多肽的多核酸的重组载体。
11.根据权利要求1至10中任一项的重组原核或真核细胞在脂肪酸、碳氢化合物、脂肪醛或脂肪醇的生产中的用途。
12.生产脂肪酸的方法，包含以下步骤：
(i)提供根据权利要求1至7中任一项的重组原核或真核细胞；和
(ii)在适合的培养条件下培养所述重组原核或真核细胞。
13.生产碳氢化合物，例如中链碳氢化合物或短链碳氢化合物的方法，包含实施根据权利要求12的生产脂肪酸的方法，进一步包含所述脂肪酸的还原、氢化、脱羧或脱碳。
14.生产脂肪醇或脂肪醛，例如中链脂肪醇或醛或者短链脂肪醇或醛的方法，包含实施根据权利要求12的生产脂肪酸的方法，进一步包含还原所述脂肪酸。
15.根据权利要求11的用途或根据权利要求13或14的方法，其中所述脂肪酸为短链脂肪酸(SCFA)或中链脂肪酸(MCFA)，优选C6-C12脂肪酸。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP16158805 | 2016-03-04 | ||
| EP16158805.8 | 2016-03-04 | ||
| PCT/EP2017/053811 WO2017148727A1 (en) | 2016-03-04 | 2017-02-20 | Method of producing fatty acids |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108779444A true CN108779444A (zh) | 2018-11-09 |
Family
ID=55699339
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201780014909.4A Pending CN108779444A (zh) | 2016-03-04 | 2017-02-20 | 生产脂肪酸的方法 |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US10648043B2 (zh) |
| EP (1) | EP3423573B1 (zh) |
| CN (1) | CN108779444A (zh) |
| AU (1) | AU2017225545A1 (zh) |
| BR (1) | BR112018016151A2 (zh) |
| ES (1) | ES2841298T3 (zh) |
| PT (1) | PT3423573T (zh) |
| WO (1) | WO2017148727A1 (zh) |
| ZA (1) | ZA201806415B (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112063608A (zh) * | 2020-08-27 | 2020-12-11 | 浙江工业大学 | 一种脂肪酸光脱羧酶突变体及其在l-草铵膦合成中的应用 |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3423573B1 (en) | 2016-03-04 | 2020-11-11 | Total Raffinage Chimie | Method of producing fatty acids |
| CN112410355B (zh) * | 2020-11-23 | 2022-03-25 | 昆明理工大学 | 一种酰基辅酶a氧化酶2基因rkacox2及其应用 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP3423573B1 (en) | 2016-03-04 | 2020-11-11 | Total Raffinage Chimie | Method of producing fatty acids |
-
2017
- 2017-02-20 EP EP17709371.3A patent/EP3423573B1/en active Active
- 2017-02-20 BR BR112018016151A patent/BR112018016151A2/pt not_active Application Discontinuation
- 2017-02-20 PT PT177093713T patent/PT3423573T/pt unknown
- 2017-02-20 WO PCT/EP2017/053811 patent/WO2017148727A1/en active Application Filing
- 2017-02-20 CN CN201780014909.4A patent/CN108779444A/zh active Pending
- 2017-02-20 US US16/072,101 patent/US10648043B2/en not_active Expired - Fee Related
- 2017-02-20 ES ES17709371T patent/ES2841298T3/es active Active
- 2017-02-20 AU AU2017225545A patent/AU2017225545A1/en not_active Abandoned
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2018
- 2018-09-26 ZA ZA2018/06415A patent/ZA201806415B/en unknown
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| US20130029387A1 (en) * | 2011-07-27 | 2013-01-31 | Iowa State University Research Foundation, Inc. | Materials and methods for using an acyl-acyl carrier protein thioesterase and mutants and chimeras thereof in fatty acid synthesis |
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| CN112063608A (zh) * | 2020-08-27 | 2020-12-11 | 浙江工业大学 | 一种脂肪酸光脱羧酶突变体及其在l-草铵膦合成中的应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| BR112018016151A2 (pt) | 2019-01-29 |
| ZA201806415B (en) | 2019-07-31 |
| US20190040477A1 (en) | 2019-02-07 |
| AU2017225545A1 (en) | 2018-08-16 |
| US10648043B2 (en) | 2020-05-12 |
| ES2841298T3 (es) | 2021-07-08 |
| PT3423573T (pt) | 2021-01-20 |
| EP3423573A1 (en) | 2019-01-09 |
| EP3423573B1 (en) | 2020-11-11 |
| WO2017148727A1 (en) | 2017-09-08 |
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