CN107075450B - 在红冬孢酵母属和红酵母属物种中高效产生多不饱和脂肪酸(pufa)的方法 - Google Patents

在红冬孢酵母属和红酵母属物种中高效产生多不饱和脂肪酸(pufa)的方法 Download PDF

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CN107075450B
CN107075450B CN201580060087.4A CN201580060087A CN107075450B CN 107075450 B CN107075450 B CN 107075450B CN 201580060087 A CN201580060087 A CN 201580060087A CN 107075450 B CN107075450 B CN 107075450B
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C·M·许
纪良辉
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Abstract

本发明涉及真菌生物技术领域，更特别是在选自红冬孢酵母属和红酵母属的真菌宿主中产生多不饱和脂肪酸(PUFA)的遗传工程方法。

Description

在红冬孢酵母属和红酵母属物种中高效产生多不饱和脂肪酸 (PUFA)的方法
相关申请的交叉引用
本申请涉及并要求2014年9月8日提交的美国临时专利申请系列号62/047,300的优先权。该申请整体援引加入本文。
序列提交
本申请与电子格式的序列表一起提交。序列表题为2577237PCTSequenceListing.txt，2015年7月2日创建，并且大小为321kb。电子格式的序列表的信息整体援引加入本文。
发明背景
本发明涉及真菌生物技术领域，更特别是在选自红冬孢酵母属(Rhodospordium)和红酵母属(Rhodotorula)的真菌宿主中产生ω-3多不饱和脂肪酸(PUFA)的遗传工程方法。
本文中用来说明本发明的背景或提供关于实践的额外细节的出版物和其他材料援引加入本文，并且为了方便，分别分组在参考文献中。
Omega-3脂肪酸(也称作ω-3脂肪酸或n-3脂肪酸)是指α-亚麻酸(ALA)[(9Z,12Z,15Z)-9,12,15-十八碳三烯酸]、EPA(二十碳五烯酸，或[(5Z,8Z,11Z,14Z,17Z)-5,8,11,14,17-二十碳五烯酸])和DHA[二十二碳六烯酸，或(4Z,7Z,10Z,13Z,16Z,19Z)-二十二碳-4,7,10,13,16,19-六烯酸]。动物ω-3EPA和DHA脂肪酸的常见来源包括鱼油、卵油、乌贼油和磷虾油，而一些植物油如来自沙棘籽和浆果、藻类细胞、亚麻籽、奇亚籽和火麻仁的油包含高水平的ALA。
亚油酸[(9Z,12Z)-9,12-十八碳二烯酸]、γ-亚麻酸(GLA，或all-cis-6,9,12-十八碳三烯酸)和花生四烯酸[(5Z,8Z,11Z,14Z)-5,8,11,14-二十碳四烯酸])是ω-6脂肪酸。GLA是主要在基于植物的油如琉璃苣种子油、月见草油和黑醋栗种子油中发现的ω-6脂肪酸。
ω-3脂肪酸对于正常代谢是至关重要的。据认为ω-3是必需脂肪酸，即，不可以通过人体合成，除了哺乳动物具有有限能力，当饮食包括短链ω-3脂肪酸ALA时，以形成更重要的长链ω-3脂肪酸EPA，然后从EPA，以甚至更低的效率形成最重要的DHA。现在接受ω-3多不饱和脂肪酸，特别是EPA和DHA在人类健康的许多方面起重要作用。但是，过度捕捞和关于海洋环境污染的担心表明需要开发极长链多不饱和脂肪酸(VLC-PUFA)如EPA和DHA的可选的可持续来源[1]。据认为ω-6脂肪酸是必需脂肪酸：它们对于人类健康是必需的。连同ω-3脂肪酸，ω-6脂肪酸在脑功能以及正常生长和发育中起到至关重要的作用。ω-3脂肪酸和ω-6脂肪酸有助于刺激皮肤和毛发生长，保持骨骼健康，调节代谢，以及维持生殖系统[2]。一些初步临床研究表明GLA可能可用于糖尿病性神经病、类风湿性关节炎、过敏、湿疹、高血压(Hypertension)、绝经期症状等。据认为ω-6和ω-3必需脂肪酸的膳食摄入比例对于人体健康很重要[3]。
到目前为止已报道大量产油微生物。它们产生的油(常称作单细胞油(SCO))与植物的相似，并且可以用于生产生物柴油、食品和工业产品[4-6]。SCO现在在市场上被广泛接受，并且越来越意识到PUFA如γ-亚麻酸(GLA)、花生四烯酸(ARA)、DHA和EPA的健康益处。ARA和DHA还已在世界许多地方用于强化婴儿配方。鱼油是DHA和EPA的丰富来源，并且有限数量的植物油料种子是其他PUFA的良好来源。海洋原生生物和沟鞭藻类如破囊壶菌属(Thraustochytrium)、裂殖壶菌属(Schizochytrium)和Crypthecodinium的物种是DHA的丰富来源，而微藻如褐指藻属(Phaeodactylum)和Monodus是EPA的良好来源。低等真菌被孢霉属(Mortierella)的物种在脂质级分中积累高百分比的ARA[7]。
虽然酵母解脂耶氏酵母(Yarrowia lipolytica)可能作为SCO的生物工程宿主享有悠久的研究和开发历史[8-12]，但是圆红冬孢酵母(Rhodosporidium toruloides)(也称作Rhodotorula glutenis)已吸引越来越多的关注，这是由于其能够以快速生长速度进行较高的细胞密度发酵，高效产生具有>67％(w/w干细胞质量)油含量的细胞团块[13-16]。
柄锈菌亚门(Pucciniomycotina)是担子菌门(Basidiomycota)中的真菌亚门[17]。其拥有许多具有重要工业应用的物种。例如，红冬孢酵母属(Rhodosporidium)和锁掷酵母属(Sporidiobolus)中的许多物种如圆红冬孢酵母(也称作瘦弱红酵母(Rhodotorulagracilis)、Rhodosporidium glutinis、粘红酵母(Rhodotorula glutinis)、Torulakoishikawensis和Torula rubescens)和赭色掷孢酵母(Sporobolomyces salmonicolor)是能够高密度发酵的富含油的单细胞酵母[6,18]。这些物种拥有巨大潜力作为宿主用于产生长链烃，如三酰甘油(TAG，或脂肪)、脂肪酸酯(生物柴油)、脂肪醇、醇、内酯、萜类化合物和维生素[14,19-21]。
红冬孢酵母属和红酵母属基因组是高度富含GC的，已发现这深刻影响遗传转化和蛋白表达[22-24]。代谢工程是提高植物和微生物中的代谢物生成的有效技术。在ω-3脂肪酸的生物工程方面，在植物和产油酵母中表达各种去饱和酶和延长酶对于PUFA的产生至关重要[25]。GLA通过Δ6-去饱和酶合成自亚油酸(LA；C18:2Δ9,12cis)。红花(Carthamustinctorius)的种子油包含高LA，并且已通过用来自高山被孢霉(Mortierella alpina)和异枝水霉(Saprolegnia diclina)的Δ6-去饱和酶转化进行修饰以分别获得超过50％(v/v)的GLA[26]。
ALA和GLA是产生较长链ω-3脂肪酸如花生四烯酸(AA)、EPA和二十二碳六烯酸(DHA)的前体[7,27]。因此，以高容积生产率产生高水平的ALA和GLA的能力对于圆红冬孢酵母中较长链PUFA的生物工程至关重要。因此，需要开发产生高水平的ALA和GLA(然后在真菌物种中可用于产生较长链PUFA)的红冬孢酵母属和红酵母属的真菌物种。
发明概述
本发明涉及真菌生物技术领域，更特别是在选自红冬孢酵母属和红酵母属的真菌宿主中产生ω-3多不饱和脂肪酸(PUFA)的遗传工程方法。
在第一方面，本发明提供一种真菌宿主，其具有以所述真菌宿主细胞中总脂肪酸的至少9％量存在的α-亚麻酸(ALA)。在一实施方案中，所述真菌宿主是红冬孢酵母属的物种。在另一实施方案中，所述真菌宿主是红酵母属的物种。在一些实施方案中，所述真菌宿主具有减少的天然醛脱氢酶(ALD)活性，所述天然醛脱氢酶(ALD)使用脂肪酸醛作为底物。所述天然ALD由天然ALD基因编码。在一实施方案中，所述天然ALD基因编码具有SEQ ID NO:3中示出的氨基酸序列的醛脱氢酶(ALD)。在另一实施方案中，所述天然ALD基因编码具有SEQ ID NO:3中示出的氨基酸序列的75％、80％、85％、90％、95％、98％或99％的ALD。在一实施方案中，所述天然ALD基因具有SEQ ID NO:1中示出的基因组核苷酸序列。在另一实施方案中，所述天然ALD基因具有SEQ ID NO:2中示出的cDNA核苷酸序列。在另一实施方案中，所述天然ALD基因具有这样的核苷酸序列，其具有SEQ ID NO:1或SEQ ID NO:2中示出的核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一些实施方案中，减少的天然ALD活性是由减少的天然ALD基因表达引起的。减少的表达可以由任何遗传或表观遗传机制引起。在一实施方案中，减少的表达是由RNAi机制如siRNA、shRNA、miRNA等引起的。在另一实施方案中，减少的表达是由人工转录阻遏物引起的。在另一实施方案中，减少的表达是由反义机制引起的。在一实施方案中，减少的表达是由有义抑制引起的。在另一实施方案中，减少的活性是由天然基因的突变引起的。在一实施方案中，所述突变可以是导致活性减少的取代、缺失、插入、添加或倒位等。在另一实施方案中，所述突变可以由同源重组引起。在另一实施方案中，所述突变可以由T-DNA或转座子插入引起。
在第二方面，本发明提供一种真菌宿主，其具有以所述真菌宿主细胞中总脂肪酸的至少49％的量存在的α-亚麻酸(ALA)。在一实施方案中，所述真菌宿主是红冬孢酵母属的物种。在另一实施方案中，所述真菌宿主是红酵母属的物种。在一些实施方案中，如本文所述，所述真菌宿主具有减少的天然醛脱氢酶(ALD)活性。在其他实施方案中，所述真菌宿主的基因组已被修饰以稳定地包括编码参与脂肪酸生物合成的蛋白的两个或更多个基因。这类蛋白的实例为酰基-CoA δ-12去饱和酶、硬脂酰-CoA-δ-9-去饱和酶、ω-3去饱和酶、脂肪酸延长酶、乙酰基-CoA羧化酶(ACC)、ATP:柠檬酸裂解酶(ACL)、二酰甘油酰基转移酶(DGA)或苹果酸酶(MAE)。在一些实施方案中，这类基因的编码序列已被修饰以包含至少55％G和C含量，优选60％-70％G和C含量。在其他实施方案中，至少70％的密码子在第三位具有C或G。
在一实施方案中，ATP:柠檬酸裂解酶(ACL)具有SEQ ID NO:88中示出的氨基酸序列。在另一实施方案中，这种ATP:柠檬酸裂解酶(ACL1)由具有SEQ ID NO:86中示出的核苷酸序列的基因组DNA核酸编码。在另一实施方案中，这种ATP:柠檬酸裂解酶(ACL1)由具有SEQ ID NO:87中示出的核苷酸序列的cDNA核酸编码。在另一实施方案中，ATP:柠檬酸裂解酶(ACL1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，二酰甘油酰基转移酶(DGA1)具有SEQ ID NO:82中示出的氨基酸序列。在另一实施方案中，这种二酰甘油酰基转移酶(DGA1)由具有SEQ ID NO:80中示出的核苷酸序列的基因组DNA编码。在另一实施方案中，这种二酰甘油酰基转移酶(DGA1)由具有SEQ ID NO:81中示出的核苷酸序列的核酸编码。在另一实施方案中，二酰甘油酰基转移酶(DGA1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，苹果酸酶(MAE1)具有SEQ ID NO:85中示出的氨基酸序列。在另一实施方案中，这种苹果酸酶(MAE1)由具有SEQ ID NO:83中示出的核苷酸序列的基因组DNA核酸编码。在另一实施方案中，这种苹果酸酶(MAE1)由具有SEQ ID NO:84中示出的核苷酸序列的核酸编码。在另一实施方案中，苹果酸酶(MAE1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在另一方面，本发明提供一种真菌宿主，其具有以所述真菌宿主细胞中总脂肪酸的至少30％的量存在的γ-亚麻酸(GLA)。在一实施方案中，所述真菌宿主是红冬孢酵母属的物种。在另一实施方案中，所述真菌宿主是红酵母属的物种。在一些实施方案中，如本文所述，所述真菌宿主具有减少的天然醛脱氢酶(ALD)活性。在其他实施方案中，所述真菌宿主的基因组已被修饰以稳定地表达编码参与脂肪酸生物合成的蛋白的两个或更多个额外基因。这类蛋白的实例为酰基-CoA δ-12去饱和酶、硬脂酰-CoA-δ-9-去饱和酶、酰基-CoAδ-6去饱和酶、脂肪酸延长酶、乙酰基-CoA羧化酶(ACC)、ATP:柠檬酸裂解酶(ACL)、二酰甘油酰基转移酶(DGA)或苹果酸酶(MAE)。在一些实施方案中，这类基因的编码序列已被修饰以包含至少55％G和C含量，优选60％-70％G和C含量。在其他实施方案中，至少70％的密码子在第三位置具有C或G。
在一实施方案中，乙酰基-CoA羧化酶(ACC1)具有SEQ ID NO:91中示出的氨基酸序列。在另一实施方案中，这种乙酰基-CoA羧化酶(ACC1)由具有SEQ ID NO:89中示出的核苷酸序列的基因组DNA核酸编码。在另一实施方案中，这种乙酰基-CoA羧化酶(ACC1)由具有SEQ ID NO:90中示出的核苷酸序列的cDNA核酸编码。在另一实施方案中，乙酰基-CoA羧化酶(ACC1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，酰基-CoA δ-12去饱和酶具有SEQ ID NO:5和94中示出的氨基酸序列。在另一实施方案中，这种酰基-CoA δ-12去饱和酶由具有SEQ ID NO:4、92和NO:93中示出的核苷酸序列的核酸编码。在另一实施方案中，酰基-CoA δ-12去饱和酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，硬脂酰-CoA-δ-9-去饱和酶具有SEQ ID NO:8中示出的氨基酸序列。在一实施方案中，这种硬脂酰-CoA-δ-9-去饱和酶由具有SEQ ID NO:6中示出的核苷酸序列的基因组核酸编码。在另一实施方案中，这种硬脂酰-CoA-δ-9-去饱和酶由具有SEQ IDNO:7中示出的核苷酸序列的核酸编码。在另一实施方案中，硬脂酰-CoA-δ-9-去饱和酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，ω-3去饱和酶具有SEQ ID NO:10中示出的氨基酸序列。在一实施方案中，这种ω-3去饱和酶由具有SEQ ID NO:9中示出的核苷酸序列的核酸编码。在一额外实施方案中，ω-3去饱和酶具有SEQ ID NO:12中示出的氨基酸序列。在一实施方案中，后者这种ω-3去饱和酶由具有SEQ ID NO:11中示出的核苷酸序列的核酸编码。在另一实施方案中，ω-3去饱和酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，酰基-CoA δ-6去饱和酶具有SEQ ID NO:96和98中示出的氨基酸序列，其中DNA编码的序列包含至少55％G和C，优选60％-70％G和C。在其他实施方案中，至少70％的密码子在第三位具有C或G。在一实施方案中，酰基-CoA δ-6去饱和酶由SEQ IDNO:95和97中示出的核酸编码。
在一实施方案中，脂肪酸延长酶具有SEQ ID NO:101和104中示出的氨基酸序列。在另一实施方案中，这种脂肪酸延长酶由具有SEQ ID NO:99、100、102和103中示出的核苷酸序列的核酸编码。在另一实施方案中，脂肪酸延长酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一些实施方案中，将本文描述的已稳定并入真菌基因组的基因可操作地连接至允许在红冬孢酵母属和红酵母属的物种中高效表达的启动子。每个并入基因的启动子可以相同或不同。在一些实施方案中，所述启动子是在红冬孢酵母属和红酵母属的物种中发现的启动子。合适的启动子包括但不限于编码以下蛋白的以下基因的启动子：甘油醛3-磷酸脱氢酶(GPD)、酰基-CoA载体蛋白(ACP)、脂肪酸去饱和酶、翻译延伸因子(TEF)、丙酮酸脱羧酶(PDC)、烯醇化酶(2-磷酸甘油酸脱水酶)(ENO)、肽基脯氨酰异构酶(PPI)、乙酰基-CoA羧化酶(ACC)或转醛醇酶。在其他实施方案中，本文所述基因还包括mRNA转录终止子，其可以是在任何真核物种和它们的DNA病毒中发现的mRNA转录终止子。
在另一方面，本发明提供一种制备ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使本文所述真菌宿主细胞在适合产生PUFA的条件下生长。可以使用具有至少5％碳源如葡萄糖、甘露糖、甘油、蔗糖的任何培养基。培养基的实例是MinLG培养基，其包含30-100g葡萄糖、1.5g酵母提取物、0.5g(NH4)2SO4、2.05g K2HPO4、1.45g KH2PO4、0.6g MgSO4、0.3g NaCl、10mg CaCl2、1mg FeSO4、0.5mg ZnSO4、0.5mg CuSO4、0.5mg H3BO4、0.5mg MnSO4、0.5mg NaMoO4(每升)。将培养基pH调整至6-7。在25℃-32℃下进行细胞培养。
附图简述
图1A-1E示出本发明中使用的构建体中的T-DNA结构。所有二元载体均具有相同的pPZP200骨架[28]。图1A：pEC3GPD-GUS。图1B：pEC3Pxxx-HPT3。图1C：pRH2034。图1D：pRHDGA1和pRHMAE1。图1E：pRH201。LB：T-DNA的左边界；RB：T-DNA的右边界；Pgpd：Umgpd1的595bp启动子；PGPD1：RtGPD1的795bp启动子；Pxxx：各种启动子；hpt-3：基于圆红冬孢酵母(R.toruloides)中的密码子使用偏好的密码子优化的潮霉素抗性基因；GUS：大肠杆菌(E.coli)β-葡糖醛酸酶基因；T35S：花椰菜花叶病毒35S基因的终止子；Tnos：根癌农杆菌(A.tumefaciens)胭脂氨酸合成酶基因的终止子。示出质粒中的独特限制性酶切割位点。
图2A-2C示出本发明中使用的构建体中的T-DNA结构。所有二元载体均具有相同的pPZP200骨架[28]。图2A：pRHE001。图2B：pRHE002。图2C：pRHE003。LB：T-DNA的左边界；RB：T-DNA的右边界；Pgpd：Umgpd1的595bp启动子；PGPD1：RtGPD1的795bp启动子；hpt-3：基于圆红冬孢酵母中的密码子使用偏好的密码子优化的潮霉素抗性基因；T35S：花椰菜花叶病毒35S基因的终止子；Tnos：根癌农杆菌胭脂氨酸合成酶基因的终止子。示出质粒中的独特限制性酶切割位点。
图3A-3C示出本发明中使用的构建体中的T-DNA结构。所有二元载体均具有相同的pPZP200骨架[28]。图3A：pRHE004。图3B：pRHE005。图3C：pRHE006。LB：T-DNA的左边界；RB：T-DNA的右边界；Pgpd：Umgpd1的595bp启动子；PGPD1：RtGPD1的795bp启动子；hpt-3：基于圆红冬孢酵母中的密码子使用偏好的密码子优化的潮霉素抗性基因；T35S：花椰菜花叶病毒35S基因的终止子；Tnos：根癌农杆菌胭脂氨酸合成酶基因的终止子。示出质粒中的独特限制性酶切割位点。
图4A-4D示出通过正向遗传学鉴定的突变菌株。图4A：WT和针对浅蓝菌素选择的T-DNA突变体(RCM)中平均ALA(C18:3n＝9)水平的比较。图4B：个体RCM突变体中的相对ALA水平。图4C：WT和针对四氮唑紫(tetrazeolium violet)选择的突变体中脂质积累的比较。图4D：WT和针对荧光染料尼罗红选择的突变体中脂质积累的比较。
图5A-5F示出圆红冬孢酵母中ALD1的反向遗传学研究。图5A：ALD1及其缺失策略的示意图。用于ALD1缺失的同源序列长度为730bp和829bp，在翻译起始密码子的-185至+535和+1948至+2776之间。图5B：Δald1的DNA印迹分析。将地高辛标记的DNA序列(图5A中标记的ALD1R)用作针对用HincII消化的基因组DNA的探针。图5C：在第3和4天WT和Δald1中的相对脂质收率。图5D：WT和Δald1中的相对α-亚麻酸(ALA)收率。**代表通过统计t-检验非常显著的差异(P<0.01)。图5E：WT和Δald1的干细胞生物量(Biomass)和ALA含量(总脂肪酸的百分比，％TFA)。图5F：马铃薯葡萄糖琼脂(PDA)和YPD肉汤中培养的细胞的颜色。
图6示出来自各种担子菌类(Basidiomycotous)物种的脂肪醛脱氢酶的比对。Ab：双孢蘑菇(Agaricus bisporus)(EKM75339.1；SEQ ID NO:105)；Pc：肉质显丝菌(Phanerochaete carnosa)(EKM57674.1；SEQ ID NO:106)；Mg：球形马拉色菌(Malasseziaglobosa)(XP_001730031.1；SEQ ID NO:107)；Sr：玉米丝黑穗病菌(Sporisoriumreilianum)(CBQ71609.1；SEQ ID NO:108)；Um：玉米黑粉菌(Ustilago maydis)(XP_762570.1；SEQ ID NO:109)；Ml：松杨栅锈菌(Melampsora larici-populina)(EGG04055.1；SEQ ID NO:110)；Pg：Puccinia graminis f.sp.tritici(XP_003338710.1；SEQ ID NO:111)；Pt：小麦叶锈菌(Puccinia triticina)(XP_003338710.1；SEQ ID NO:112)；Ps：条锈病菌(Puccinia striiformis)(基因组基因座CQM_00777.1；SEQ ID NO:113)；Mv：花药黑粉菌(Microbotryum violaceum)(基因组基因座MVLG_02667.1；SEQ ID NO:114)；Rg2:R.glutinis ATCC 204091(SEQ ID NO:115)；Rt3：圆红冬孢酵母NP11(EMS18750.1；SEQ IDNO:116)；Rt1：圆红冬孢酵母ATCC 10657(SEQ ID NO:117)。[注意：序列表仅包括SEQ IDNO:112、113、115和117的部分序列。参见完整序列的图。]
图7A-7F示出DGA1和MAE1在提高脂质积累中的效果。图7A：圆红冬孢酵母ATCC10657中DGA1和MAE1的定量RT-PCR分析。将工程化突变体(DGA1和MAE1)中的基因表达针对WT菌株中的归一化。图7B：野生型和两个工程化菌株(WT、DGA1和MAE1)中的相对脂质收率。将脂质量针对WT中的归一化。图7C：上述菌株中的脂肪酸谱。3-天生物过程之后从上述菌株提取细胞内脂质。图7D：上述菌株中不饱和脂肪酸的组成。％TFA代表总脂肪酸的百分比。图7E：DGA1缺失的示意图。图7F：Δdga1的DNA印迹分析。将野生型和候选突变体的基因组DNA用HincII消化并针对如图7E中标记的DIG-标记DGA1L DNA片段探测。
图8示出不同圆红冬孢酵母(R.glutinis)菌株中的脂肪酸谱。Rt1：圆红冬孢酵母ATCC 10657；Rt2：圆红冬孢酵母ATCC 10788；Rg1：R.glutinis ATCC 90781；Rg2：R.glutinis ATCC 204091。如以前所述[21]在脂质积累培养基(MinLG)中培养圆红冬孢酵母(R.glutinis)，有一些修改。MinLG培养基包含30g葡萄糖、1.5g酵母提取物、0.5g(NH4)2SO4、2.05g K2HPO4、1.45g KH2PO4、0.6g MgSO4、0.3g NaCl、10mg CaCl2、1mg FeSO4、0.5mgZnSO4、0.5mg CuSO4、0.5mg H3BO4、0.5mg MnSO4、0.5mg NaMoO4(每升)。将培养基pH调整至6.1。在28℃下进行细胞培养4天，持续振荡(250rpm)。
图9A和9B示出高ALA菌株的工程化过程。图9A：野生型和不同去饱和酶工程菌株中的ALA含量。图9B：ald1空菌株(ald1e)以及包含RtGPD1::MaFAd2-2和RtGPD1::LuFAD3-2基因盒的ald1e菌株中的ALA含量。
图10示出ALD1和C-末端截短的ALD1(ALD1n)的酶测定。将反应与20mM Tris-Cl缓冲液(pH8.0)、1.5mM NAD+或NADP+、1.0mM十二醛以及10μl纯化的酶混合。在25℃下进行测定2min。
图11A-11D示出圆红冬孢酵母ATCC 10657中的δ-12去饱和酶基因(RtFAD2)的表征。图11A：WT和FAD2敲除菌株(fad2Δ)的DNA印迹分析。将总DNA用PstI消化，并且利用地高辛标记的FAD2的正确同源PCR片段杂交印迹(FAD2R，图11C)。图11B：一式三份进行的WT和fad2Δ的脂肪酸含量谱。图11D：在补充和未补充LA(C18:2)的YNB培养基中培养的WT和fad2Δ的脂肪酸含量谱。
图12A-12D示出圆红冬孢酵母ATCC 10657中的ELO1和ELO2缺失突变体的表征。(elo1Δ和elo2Δ)(图12A)和(图12C)elo1Δ和elo2Δ的DNA印迹分析。将总DNA用PvuI消化，并且分别针对地高辛标记的ELO1R和ELO2L杂交。(图12B)和(图12D)elo1Δ和elo2Δ的脂肪酸含量谱。
图13A-13E示出图14中使用的载体的示意图。图13A：FAD1(δ-9-油酸去饱和酶，ELO1)过量表达构建体。图13B：FAD2(δ-12去饱和酶)过量表达构建体。图13C：ω-3去饱和酶(δ-15去饱和酶)过量表达构建体。图13D：FAD1和FAD2双基因过量表达载体。图13E：FAD1、FAD2和ω-3去饱和酶三基因过量表达载体。LB：T-DNA的左边界；RB：T-DNA的右边界；RgGPD1：源自Rhodotorula grammis WP1的685bp GPD1启动子；RtGPD1：源自圆红冬孢酵母ATCC 10657的795bp GPD1启动子；hpt-3：基于圆红冬孢酵母中的密码子使用偏好的密码子优化的潮霉素抗性基因；TSV40：猿猴空泡病毒40大T抗原基因终止子；T35S：花椰菜花叶病毒35S基因的终止子；Tnos：根癌农杆菌胭脂氨酸合成酶基因的终止子；RtENO1：来自圆红冬孢酵母ATCC 10657的445bp版本ENO1基因启动子；RtACC1：来自圆红冬孢酵母ATCC 10657的ACC1基因启动子的805bp版本；MaFAD2-2是SEQ ID NO:4，并且VfFAD3-2是SEQ ID NO:11。
图14：工程化菌株的脂肪酸含量谱。将3个独立的转化体用于每个分析，除了Δald1-OMA2，对其分析总计18个独立的转化体。将所有转化体在RL2培养基中发酵5天。WT：野生型圆红冬孢酵母ATCC 10657菌株；Δald1-：将构建体转化入ADL1敲除突变体；构建体在图13中示出。OEL1：图13A；MA：图13B；AF3：图13C；OM2：图13D；OMA2：图13E。
图15A-15D示出圆红冬孢酵母ATCC 10657中推测的ATP-柠檬酸裂解酶基因(RtACL1)的表征。图15A：ACL1基因及其缺失策略的示意图。图15B：敲除菌株acl1Δ的DNA印迹分析。将总DNA用PvuI消化，并且针对地高辛标记的ACL1L杂交。M：地高辛标记的DNA分子量标记VII(Roche Diagnosis,USA)。图15C：ACL1破坏对生物质、脂质收率、脂质含量和残余葡萄糖的影响。图15D：Wt和acl1Δ菌株的脂肪酸含量谱。
发明详述
本发明涉及真菌生物技术领域，更特别是在选自红冬孢酵母属和红酵母属的真菌宿主中产生多不饱和脂肪酸(PUFA)的遗传工程方法。
除非另有定义，本文使用的所有技术和科学术语具有与本发明所属领域技术人员通常理解的相同意义。
如本文所用，“ALD1”是使用脂肪酸醛作为底物的醛脱氢酶。
如本文所用，“等位基因”是指基因座位的任何一种或多种可选形式，所有所述等位基因均涉及一种性状或特征。在二倍体细胞或生物体中，给定基因的两个等位基因占据一对同源染色体上的相应基因座。
如本文中在RNAi的上下文中所用，“dsRNA”或“RNAi分子”是指这样的化合物，其能够下调或减少基因的表达或这样的基因产物的活性至足以达到期望的生物或生理效果的程度。如本文所用，术语“dsRNA”或“RNAi分子”是指dsRNA、siRNA、shRNA、ihpRNA、合成shRNA、miRNA中的一种或多种。
当指主题RNAi方法抑制的基因时，术语“下调”是指在一个或多个RNAi构建体的存在下，与不存在这样的RNAi构建体的水平相比时基因的表达水平降低。术语“下调”在本文中用来表示靶基因表达降低1-100％。例如，表达可以减少约5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％或99％。
关于基因序列，术语“表达”是指基因的转录以及在适当时将所得的mRNA转录物翻译为蛋白。因此，从上下文会清楚，蛋白编码序列的表达导致编码序列的转录和翻译。
如本文所用，“基因”是指涵盖与基因产物表达相关的5′启动子区、任何内含子和外显子区以及与基因产物表达相关的3′或5′非翻译区的核酸序列。
术语“基因沉默”是指基因表达的抑制，例如，转基因、异源基因和/或内源基因表达。基因沉默可以通过影响转录的过程和/或通过影响转录后机制的过程介导。基因沉默可以是等位基因特异性的，其中发生基因的一个等位基因的特异性沉默。
如本文所用，“基因型”是指细胞或生物体的遗传组成。
当参考核酸部分使用时，术语“异源”或“外源”表示所述核酸包含在自然中互相相同的关系中未发现的两个或更多个子序列。例如，所述核酸通常重组产生，具有来自无关基因的两个或更多个序列排列以产生新的功能核酸，例如来自一个来源的启动子和来自另一来源的编码区。相似地，异源或外源蛋白表示所述蛋白包含在自然中互相相同的关系中未发现的两个或更多个子序列(例如，融合蛋白)。
如本文所用，术语“同系物”是指通过来自共同祖先DNA序列的血统与第二基因相关的基因。术语同系物可以应用于通过物种形成事件分离的基因之间的关系(直系同源物)或者通过遗传复制事件分离的基因之间的关系(旁系同源物)。术语同系物一般用来指所有物种。
如本文所用，“表型”是指细胞或生物体的可检测的特征，所述特征是基因表达的表现。
术语“多核苷酸”、“核酸”和“核酸分子”在本文中可交换地用来指核苷酸的聚合物，其可以是天然或合成的核苷酸和/或核苷的线性和顺序阵列，包括脱氧核糖核酸、核糖核酸和它们的衍生物。其包括染色体DNA、自我复制质粒、DNA或RNA的感染性聚合物以及发挥主要结构作用的DNA或RNA。除非另有说明，核酸或多核苷酸以5’至3’方向从左至右书写。通过它们普遍接受的单字母代码引用核苷酸。数字范围包括定义范围的数字。
术语“多肽”、“肽”和“蛋白”在本文中可交换地用来指氨基酸残基的聚合物。该术语应用于其中一个或多个氨基酸残基是相应天然存在的氨基酸的人工化学类似物的氨基酸聚合物，以及天然存在的氨基酸聚合物。氨基酸可以通过它们常用的三字母或单字母符号引用。氨基酸序列分别以氨基至羧基方向从左至右书写。数字范围包括定义范围的数字。
如本文所用，“可操作的连接”或“可操作地连接”或“操作性地连接”理解为表示，例如，启动子和待表达的核酸以及其他调节元件(如果适当)如终止子的顺序排列为这样的方式，每个调节元件可以履行其在重组表达核酸以产生dsRNA中的功能。这不必要求在化学意义上直接连接。遗传控制序列如增强子序列还可以从距离有些遥远的位置发挥它们对靶序列的功能，或者实际上从其他DNA分子(顺式或反式定位)。优选排列是其中待重组表达的核酸序列位于充当启动子的序列下游的排列，从而两个序列互相共价键合。如本领域公知的，调节或控制序列可以位于核苷酸序列的5’侧或核苷酸序列的3’侧。
术语“约”或“大约”表示在值的统计学上有意义的范围内。这样的范围可以在一个数量级内，优选在给定值或范围的50％内，更优选在20％内，更优选在10％内，并且甚至更优选在5％内。术语“约”或“大约”所涵盖的允许偏差取决于研究的具体系统，并且本领域技术人员可以容易地理解。
如本文所用，术语“序列相同性”、“序列相似性”或“同源性”用来描述两个或更多个核苷酸序列之间的序列关系。两个序列之间的“序列相同性”的百分比是通过在比较窗口上比较两个最佳比对的序列如提到的SEQ ID NO:的全长来确定的，其中为了两个序列的最佳比对，比较窗口中的序列部分与参考序列(其不含添加或缺失)相比可以包含添加或缺失(即，缺口)。百分比这样计算，确定在两个序列中均存在的相同核酸碱基或氨基酸残基的位置的数目以产生匹配位置的数目，匹配位置的数目除以比较窗口中的位置总数目，并且将结果乘以100以产生序列相同性的百分比。与参考序列相比在每个位置相同的序列据说与参考序列相同，反之亦然。如果第一核苷酸序列表现出与第二或参考序列的完全互补性，则认为第一核苷酸序列在以5'至3'方向观察时是以3'至5'方向观察的第二或参考核苷酸序列的“补体”或与其互补。如本文所用，当5'至3'阅读的序列之一的每个核苷酸与3'至5'阅读时的另一序列的每个核苷酸互补时，则认为核酸序列分子表现出“完全互补性”。与参考核苷酸序列互补的核苷酸序列会表现出与参考核苷酸序列的反向互补序列相同的序列。这些术语和描述是本领域中良好定义的，并且本领域技术人员容易理解。
如本文所用，“比较窗口”或“比较的窗口”是指至少6个连续位置的概念片段，通常约50至约100个，更通常约100至约150个，其中在两个序列最佳比对之后将序列与相同数目连续位置的参考序列比较。为了两个序列的最佳比对，与参考序列(不含添加或缺失)相比比较窗口可以包含约20％或更少的添加或缺失(即缺口)。本领域技术人员应当参考用于序列比对的详细方法，如Wisconsin Genetics Software Package Release 7.0(GeneticsComputer Group,575Science Drive Madison,Wisc.,USA)。
在第一方面，本发明提供一种真菌宿主，其具有以所述真菌宿主细胞中总脂肪酸的至少9％量存在的α-亚麻酸(ALA)。在一实施方案中，所述真菌宿主是红冬孢酵母属的物种。在另一实施方案中，所述真菌宿主是红酵母属的物种。在一些实施方案中，所述真菌宿主具有减少的天然醛脱氢酶(ALD)活性，所述天然醛脱氢酶(ALD)使用脂肪酸醛作为底物。所述天然ALD由天然ALD基因编码。在一实施方案中，所述天然ALD基因编码具有SEQ ID NO:3中示出的氨基酸序列的醛脱氢酶(ALD)。在另一实施方案中，所述天然ALD基因编码具有SEQ ID NO:3中示出氨基酸序列的75％、80％、85％、90％、95％、98％或99％的ALD。在一实施方案中，所述天然ALD基因具有SEQ ID NO:1中示出的基因组核苷酸序列。在另一实施方案中，所述天然ALD基因具有SEQ ID NO:2中示出的cDNA核苷酸序列。在另一实施方案中，所述天然ALD基因具有这样的核苷酸序列，其具有SEQ ID NO:1或SEQ ID NO:2中示出核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一些实施方案中，减少的天然ALD活性是由减少的天然ALD基因表达引起的。减少的表达可以由任何遗传或表观遗传机制引起。在一实施方案中，减少的表达是由RNAi机制如siRNA、shRNA、miRNA等引起的。在另一实施方案中，减少的表达是由人工转录阻遏物引起的。在另一实施方案中，减少的表达是由反义机制引起的。在一实施方案中，减少的表达是由有义抑制引起的。在另一实施方案中，减少的活性是由天然基因的突变引起的。在一实施方案中，所述突变可以是导致活性减少的取代、缺失、插入、添加或倒位等。在另一实施方案中，所述突变可以由同源重组引起。在另一实施方案中，所述突变可以由T-DNA或转座子插入引起。
在第二方面，本发明提供一种真菌宿主，其具有以所述真菌宿主细胞中总脂肪酸的至少49％量存在的α-亚麻酸(ALA)。在一实施方案中，所述真菌宿主是红冬孢酵母属的物种。在另一实施方案中，所述真菌宿主是红酵母属的物种。在一些实施方案中，如本文所述，所述真菌宿主具有减少的天然醛脱氢酶(ALD)活性。在其他实施方案中，已将所述真菌宿主的基因组修饰以稳定地包括编码参与脂肪酸生物合成的蛋白的两个或更多个基因。这类蛋白的实例为酰基-CoA δ-12去饱和酶、硬脂酰-CoA-δ-9-去饱和酶、ω-3去饱和酶、脂肪酸延长酶、乙酰基-CoA羧化酶(ACC)、ATP:柠檬酸裂解酶(ACL)、二酰甘油酰基转移酶(DGA)或苹果酸酶(MAE)。在一些实施方案中，已将这类基因的编码序列修饰以包含至少55％G和C含量，优选60％-70％G和C含量。在其他实施方案中，至少70％的密码子在第三位具有C或G。
在一实施方案中，ATP:柠檬酸裂解酶(ACL)具有SEQ ID NO:88中示出的氨基酸序列。在另一实施方案中，这种ATP:柠檬酸裂解酶(ACL1)由具有SEQ ID NO:86中示出的核苷酸序列的基因组DNA核酸编码。在另一实施方案中，这种ATP:柠檬酸裂解酶(ACL1)由具有SEQ ID NO:87中示出的核苷酸序列的cDNA核酸编码。在另一实施方案中，ATP:柠檬酸裂解酶(ACL1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，二酰甘油酰基转移酶(DGA1)具有SEQ ID NO:82中示出的氨基酸序列。在另一实施方案中，这种二酰甘油酰基转移酶(DGA1)由具有SEQ ID NO:80中示出的核苷酸序列的基因组DNA编码。在另一实施方案中，这种二酰甘油酰基转移酶(DGA1)由具有SEQ ID NO:81中示出的核苷酸序列的核酸编码。在另一实施方案中，二酰甘油酰基转移酶(DGA1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，苹果酸酶(MAE1)具有SEQ ID NO:85中示出的氨基酸序列。在另一实施方案中，这种苹果酸酶(MAE1)由具有SEQ ID NO:83中示出的核苷酸序列的基因组DNA核酸编码。在另一实施方案中，这种苹果酸酶(MAE1)由具有SEQ ID NO:84中示出的核苷酸序列的核酸编码。在另一实施方案中，苹果酸酶(MAE1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在另一方面，本发明提供一种真菌宿主，其具有以所述真菌宿主细胞中总脂肪酸的至少30％量存在的γ-亚麻酸(GLA)。在一实施方案中，所述真菌宿主是红冬孢酵母属的物种。在另一实施方案中，所述真菌宿主是红酵母属的物种。在一些实施方案中，如本文所述，所述真菌宿主具有减少的天然醛脱氢酶(ALD)活性。在其他实施方案中，已将所述真菌宿主的基因组修饰以稳定地表达编码参与脂肪酸生物合成的蛋白的两个或更多个额外基因。这类蛋白的实例为酰基-CoA δ-12去饱和酶、硬脂酰-CoA-δ-9-去饱和酶、酰基-CoAδ-6去饱和酶、脂肪酸延长酶、乙酰基-CoA羧化酶(ACC)、ATP:柠檬酸裂解酶(ACL)、二酰甘油酰基转移酶(DGA)或苹果酸酶(MAE)。在一些实施方案中，这类基因的编码序列包含至少55％G和C含量，优选60％-70％G和C含量。在其他实施方案中，至少70％的密码子在第三位具有C或G。
在一实施方案中，乙酰基-CoA羧化酶(ACC)具有SEQ ID NO:91中示出的氨基酸序列。在另一实施方案中，这种乙酰基-CoA羧化酶(ACC1)由具有SEQ ID NO:89中示出的核苷酸序列的基因组DNA核酸编码。在另一实施方案中，这种乙酰基-CoA羧化酶(ACC1)由具有SEQ ID N:90中示出的核苷酸序列的cDNA核酸编码。在另一实施方案中，乙酰基-CoA羧化酶(ACC1)基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，酰基-CoA δ-12去饱和酶具有SEQ ID NO:5和94中示出的氨基酸序列。在另一实施方案中，这种酰基-CoA δ-12去饱和酶由具有SEQ ID NO:4、92和NO:93中示出的核苷酸序列的核酸编码。在另一实施方案中，酰基-CoA δ-12去饱和酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，硬脂酰-CoA-δ-9-去饱和酶具有SEQ ID NO:8中示出的氨基酸序列。在一实施方案中，这种硬脂酰-CoA-δ-9-去饱和酶由具有SEQ ID NO:6中示出的核苷酸序列的基因组核酸编码。在另一实施方案中，这种硬脂酰-CoA-δ-9-去饱和酶由具有SEQ IDNO:7中示出的核苷酸序列的核酸编码。在另一实施方案中，硬脂酰-CoA-δ-9-去饱和酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，ω-3去饱和酶具有SEQ ID NO:10中示出的氨基酸序列。在一实施方案中，这种ω-3去饱和酶由具有SEQ ID NO:9中示出的核苷酸序列的核酸编码。在一额外实施方案中，ω-3去饱和酶具有SEQ ID NO:12中示出的氨基酸序列。在一实施方案中，后者这种ω-3去饱和酶由具有SEQ ID NO:11中示出的核苷酸序列的核酸编码。在另一实施方案中，ω-3去饱和酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一实施方案中，酰基-CoA δ-6去饱和酶具有SEQ ID NO:96和98中示出的氨基酸序列，其中DNA编码的序列包含至少55％G和C，优选60％-70％G和C。在其他实施方案中，至少70％的密码子在第三位具有C或G。在一实施方案中，酰基-CoA δ-6去饱和酶由SEQ IDNO:95和97中示出的核酸编码。
在一实施方案中，脂肪酸延长酶具有SEQ ID NO:101和104中示出的氨基酸序列。在另一实施方案中，这种脂肪酸延长酶由具有SEQ ID NO:99、100、102和103中示出的核苷酸序列的核酸编码。在另一实施方案中，脂肪酸延长酶基因具有这样的核苷酸序列，其具有本文所述核苷酸序列的75％、80％、85％、90％、95％、98％或99％。
在一些实施方案中，将本文描述的已稳定并入真菌基因组的基因可操作地连接至允许在红冬孢酵母属和红酵母属的物种中高效表达的启动子。每个并入基因的启动子可以相同或不同。在一些实施方案中，所述启动子是在红冬孢酵母属和红酵母属的物种中发现的启动子。合适的启动子包括但不限于编码以下蛋白的以下基因的启动子：甘油醛3-磷酸脱氢酶(GPD)、酰基-CoA载体蛋白(ACP)、脂肪酸去饱和酶、翻译延伸因子(TEF)、丙酮酸脱羧酶(PDC)、烯醇化酶(2-磷酸甘油酸脱水酶)(ENO)、肽基脯氨酰异构酶(PPI)、乙酰基-CoA羧化酶(ACC)或转醛醇酶。在其他实施方案中，本文所述基因还包括mRNA转录终止子，其可以是在任何真核物种和它们的DNA病毒中发现的mRNA转录终止子。
在另一方面，本发明提供一种制备ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使本文所述真菌宿主细胞在适合产生PUFA的条件下生长。可以使用具有至少5％碳源如葡萄糖、甘露糖、甘油、蔗糖的任何培养基。培养基的实例是MinLG培养基，其包含30-100g葡萄糖、1.5g酵母提取物、0.5g(NH4)2SO4、2.05g K2HPO4、1.45g KH2PO4、0.6g MgSO4、0.3g NaCl、10mg CaCl2、1mg FeSO4、0.5mg ZnSO4、0.5mg CuSO4、0.5mg H3BO4、0.5mg MnSO4、0.5mg NaMoO4(每升)。将培养基pH调整至6-7。在25℃-32℃下进行细胞培养。
在一些实施方案中，将本文描述的已稳定并入真菌基因组的基因可操作地连接至允许在红冬孢酵母属和红酵母属的物种中高效表达的启动子。每个并入基因的启动子可以相同或不同。在一些实施方案中，所述启动子是在红冬孢酵母属和红酵母属的物种中发现的启动子。在其他实施方案中，所述启动子是在其他真菌物种中发现的启动子。合适的启动子包括但不限于编码以下蛋白的以下基因的启动子：甘油醛3-磷酸脱氢酶(GPD)、酰基-CoA载体蛋白(ACP)、脂肪酸去饱和酶、翻译延伸因子(TEF)、丙酮酸脱羧酶(PDC)、烯醇化酶(2-磷酸甘油酸脱水酶)(ENO)、肽基脯氨酰异构酶(PPI)、乙酰基-CoA羧化酶(ACC)或转醛醇酶。在其他实施方案中，本文所述基因还包括mRNA转录终止子，其可以是在任何真核物种和它们的DNA病毒中发现的mRNA转录终止子。
在一些实施方案中，合适的启动子是国际专利申请公开号WO2012/169969中描述的启动子，其整体援引加入本文。这个公开申请描述了源自在真菌中充当启动子的甘油醛磷酸脱氢酶基因(GPD1)、翻译起始因子基因(TEF1)和硬脂酰-CoA-δ9-去饱和酶基因(FAD1)的上游区的几个多核苷酸序列。这个公开申请中描述的启动子在SEQ ID NO:55-62中示出。在其他实施方案中，额外的启动子描述于2014年3月10日提交的国际专利申请号PCT/SG2014/000114中，其整体援引加入本文。在一实施方案中，启动子序列包含SEQ ID NO:63-79中任一个中示出的序列。在另一实施方案中，多核苷酸启动子序列包含SEQ ID NO:63-79中任一个的启动子序列，即，没有克隆位点的序列。
此外，本文所述启动子序列的可操作片段可以利用常规启动子筛选测定分离，并且可以筛选用于利用本文所述技术高效选择转化的真菌细胞。在一实施方案中，可操作片段长度(在本文中也称作启动子部分)为约400个碱基对至长达约1100个碱基对，从ATG密码子的-1位开始。如本文所用，“长达”是指公开的SEQ ID NO中示出的启动子的启动子部分的长度。因此，“长达”是指启动子序列的最大长度少于公开的SEQ ID NO的启动子的1100个核苷酸。
在一实施方案中，提供启动子序列，其与这些启动子序列中的任一个具有至少60％相同性。在另一实施方案中，提供启动子序列，其与这些启动子序列中的任一个具有至少70％相同性。在一额外实施方案中，提供启动子序列，其与这些启动子序列中的任一个具有至少80％相同性。在另一实施方案中，提供启动子序列，其与这些启动子序列中的任一个具有至少90％相同性。在另一实施方案中，提供启动子序列，其与这些启动子序列中的任一个具有至少95％相同性。在另一实施方案中，提供启动子序列，其与这些启动子序列中的任一个具有至少98％相同性。
待稳定并入真菌基因组的基因通常为DNA或多核苷酸构建体形式，其包含本文描述的启动子序列、本文描述的可操作连接的多肽编码序列和可操作连接的RNA转录终止子序列。在一实施方案中，可以使用在真菌物种中可操作的任何转录终止子。终止子通常位于基因的下游(3’)，在终止密码子(TGA、TAG或TAA)之后。终止子在RNA的加工和稳定性以及翻译中起到重要作用。大多数但不是全部终止子包含多腺苷酸化序列或切割位点。具体多腺苷酸化序列的实例为AAUAAA或AAUAAU。这些序列已知为近上游元件(NUE)(Nagaya et al.,2010)。NUE通常距离已知为远上游元件(FUE)的GU-富含区约30bp(Mogen et al.,1990；Mogen et al.,1992；Rothnie et al.1994)。FUE增强在多腺苷酸化序列或切割位点处加工，其通常是U-富含区中的CA或UA(Bassett,2007)。在终止子内，元件实际上增加转录RNA的稳定性(Ohme-Takagi et al.,1993；Newman et al.,1993；Gutiérrez et al.,1999)，并且还可以控制基因表达(Ingelbrecht,1989；An et al.,1989)。
包含真菌可操作启动子、蛋白编码DNA序列和真菌可操作终止子的DNA或核酸构建体在本文中还可以称作表达盒。表达盒可以包括本领域公知的其他转录调节区。在其他实施方案中，DNA或核酸构建体或表达盒进一步包含选择标记。选择标记是技术人员公知的，表达盒并入这类选择标记和启动子以驱动它们的表达，如国际专利申请公开号WO 2012/169969所述。在本发明中可以使用可操作地连接至任何合适的选择标记的任何合适的启动子。
在一实施方案中，选择标记的编码序列是天然存在或人工产生的编码序列，并且包含至少约60％GC。在第二实施方案中，选择标记的编码序列是天然存在或人工产生的编码序列，并且包含约70％GC。在第三实施方案中，选择标记的编码序列是天然存在或人工产生的编码序列，并且包含约75％GC。在一实施方案中，这类编码序列的至少约70％的三联密码子以C或G结束。在另一实施方案中，这类编码序列的超过约80％的三联密码子以C或G结束。在一实施方案中，选择标记的编码序列是至少60％GC，优选约70％GC，并且最优选约75％GC，其中至少70％的三联密码子以C或G结束，优选超过80％的三联密码子以C或G结束。在一实施方案中，这类编码序列在总丝氨酸(Ser)残基的至少约40％中包含UCG密码子。
在一些实施方案中，选择标记是重组无标记系统的部分。在一实施方案中，不含重组标记系统是Cre-lox重组无标记系统，如Zuo等人描述的[29]。这样的系统可用于产生无选择标记的转基因植物，包括转基因麻风树植物。在一些实施方案中，重组无标记系统位于植物可操作启动子以及一个或多个核酸片段之间。在这个实施方案中，如本文所述，通过重组事件去除标记基因将植物可操作启动子置于与一个或多个核酸片段的可操作连接中。
在制备核酸构建体或表达盒中，可以操纵各种DNA片段，以便提供正确方向和(如果合适)在正确阅读框中的DNA序列。为此，衔接物或接头可以用来连接DNA片段，或者可以包括其他操纵以提供方便的限制性位点，去除多余的DNA，去除限制性位点等。为了这个目的，可以包括体外诱变、引物修复、限制酶切、退火、重新取代，例如转换和颠换。
本发明的核酸还可以通过本领域已知的方法完全或部分合成，特别是如果期望提供植物偏好序列。因此，可以利用所选宿主偏好的密码子合成本发明的全部或部分核酸。例如，从特定宿主物种中表达的蛋白中最常使用的密码子可以确定物种偏好的密码子。核苷酸序列的其他修饰可能导致具有略微改变的活性的突变体。
用任何重组构建体产生许多单独转化的真菌可能是有用的，以便回收无任何位置效应的真菌。选择包含一个以上拷贝的引入的多核苷酸构建体也可能是优选的，从而获得重组分子的高水平表达。
如果在特定物种中是可能的，产生对于特定基因是纯合的真菌系可能是可取的。在一些物种中，这通过使用单孢子培养完成。通过使用这些技术，可以产生携带插入基因的单倍体系，然后自发或通过使用秋水仙碱加倍染色体数目。这产生对于插入基因是纯合的真菌，如果插入基因携带合适的选择标记用于检测携带该基因的真菌，这可以容易地测定。或者，真菌可以是自体受精的，导致产生孢子的混合物，在最简单的情况下，所述孢子的混合物由3种类型组成，对于插入基因纯合(25％)、杂合(50％)和空(25％)。虽然相对容易鉴别空真菌与那些包含基因的真菌，但是实际上可能通过DNA印迹分析鉴别纯合与杂合真菌，其中注意装载完全等量的来自混合群体DNA，并且通过来自插入基因特异性探针的信号强度鉴别杂合子。通过允许每个独立转化体自体受精来验证DNA印迹分析的结果是可取的，因为纯合性的额外证据可以通过这样的简单事实获得，如果真菌对于插入基因是纯合的，则来自自交个体的所有随后的真菌系均包含所述基因，而如果真菌对于所述基因是杂合的，则生长自自交种子的世代会包含空真菌系。因此，用简单的自交，可以选择纯合真菌系，其还可以通过DNA印迹分析证实。
产生纯合亲本系使得可能产生包含修饰的蛋白组分的杂交真菌和孢子。维持转基因纯合亲本系，每个亲本包含可操作地连接至启动子的第一或第二重组DNA序列。种植杂交作物的优势也并入这个方案，包括更多有价值的性状和杂种优势的结合。
除非另有说明，本发明的实施采用本领域技术内的化学、分子生物学、微生物学、重组DNA、遗传学、免疫学、细胞生物学、细胞培养和转基因生物学的常规技术。参见，例如，Maniatis et al.,1982,Molecular Cloning(Cold Spring Harbor Laboratory Press,Cold Spring Harbor,New York)；Sambrook et al.,1989,Molecular Cloning,2nd Ed.(Cold Spring Harbor Laboratory Press,Cold Spring Harbor,New York)；Sambrookand Russell,2001,Molecular Cloning,3rd Ed.(Cold Spring Harbor LaboratoryPress,Cold Spring Harbor,New York)；Green and Sambrook,2012,Molecular Cloning,4th Ed.(Cold Spring Harbor Laboratory Press,Cold Spring Harbor,New York)；Ausubel et al.,1992,Current Protocols in Molecular Biology(John Wiley&Sons,including periodic updates)；Glover,1985,DNA Cloning(IRL Press,Oxford)；Russell,1984,Molecular biology of plants:a laboratory course manual(ColdSpring Harbor Laboratory Press,Cold Spring Harbor,N.Y.)；Anand,Techniques forthe Analysis of Complex Genomes,(Academic Press,New York,1992)；Guthrie andFink,Guide to Yeast Genetics and Molecular Biology(Academic Press,New York,1991)；Harlow and Lane,1988,Antibodies,(Cold Spring Harbor Laboratory Press,Cold Spring Harbor,New York)；Nucleic Acid Hybridization(B.D.Hames&S.J.Higginseds.1984)；Transcription And Translation(B.D.Hames&S.J.Higgins eds.1984)；Culture Of Animal Cells(R.I.Freshney,Alan R.Liss,Inc.,1987)；Immobilized CellsAnd Enzymes(IRL Press,1986)；B.Perbal,A Practical Guide To Molecular Cloning(1984)；the treatise,Methods In Enzymology(Academic Press,Inc.,N.Y.)；MethodsIn Enzymology,Vols.154 and 155(Wu et al.eds.),Immunochemical Methods In CellAnd Molecular Biology(Mayer and Walker,eds.,Academic Press,London,1987)；Handbook Of Experimental Immunology,Volumes I-IV(D.M.Weir and C.C.Blackwell,eds.,1986)；Riott,Essential Immunology,6th Edition,Blackwell ScientificPublications,Oxford,1988；Fire et al.,RNA Interference Technology:From BasicScience to Drug Development,Cambridge University Press,Cambridge,2005；Schepers,RNA Interference in Practice,Wiley–VCH,2005；Engelke,RNA Interference(RNAi):The Nuts&Bolts of siRNA Technology,DNA Press,2003；Gott,RNAInterference,Editing,and Modification:Methods and Protocols(Methods inMolecular Biology),Human Press,Totowa,NJ,2004；Sohail,Gene Silencing by RNAInterference:Technology and Application,CRC,2004。
实施例
参考以下实施例描述本发明，所述实施例通过说明的方式提供而不是为了以任何方式限制本发明。利用本领域公知的标准技术或下文特别描述的技术。
实施例1
菌株、化学品、培养基和培养条件
圆红冬孢酵母菌株ATCC 10657和ATCC 10788；R.glutinis菌株ATCC 90781和R.glutinis ATCC 204091购自ATCC(USA)。R.graminis菌株WP1和掷孢酵母(Sporobolomyces roseus)FGSC 10293(IAM13481)获得自Fungal Genetics Stock Center(University of Missouri,USA)。根癌农杆菌菌株AGL1[30]用于根癌农杆菌(Agrobacterium tumefaceins)介导的转化(ATMT)。潮霉素B购自Roche Diagnostics(USA)。尼龙N和N+膜(Φ82mm,0.45μm)获得自GE Healthcare(Uppsala,Sweden)。浅蓝菌素(Sigma-Aldrich,USA)制备为DMSO中5mg/ml储备溶液。除非另有说明，其他化学品均购自Sigma-Aldrich。
在28℃下于YPD肉汤(1％酵母提取物、2％蛋白胨、2％葡萄糖)中或于固体马铃薯-葡萄糖琼脂(PDA)上维持红冬孢酵母属菌株。使根癌农杆菌在28℃下于液体或固体2YT培养基(1.6％胰蛋白胨、1％酵母提取物、0.5％NaCl)中生长。如以前所述[21]在30℃和持续振荡(200rpm)下于脂质积累培养基中培养圆红冬孢酵母，有一些修改。MinLG培养基包含(每升)30g葡萄糖、1.5g酵母提取物、0.5g(NH4)2SO4、2.05g K2HPO4、1.45g KH2PO4、0.6g MgSO4、0.3g NaCl、10mg CaCl2、1mg FeSO4、0.5mg ZnSO4、0.5mg CuSO4、0.5mg H3BO4、0.5mg MnSO4、0.5mg NaMoO4(pH6)。为了分析基因表达，使用氮限制变体MinLG-N，其改进自MinLG，将酵母提取物和硫酸铵的浓度分别减少至0.3和0.1g/l。在30℃和持续振荡(200rpm)下进行脂质积累过程。
实施例2
DNA构建体
使用的寡核苷酸在表1中列出。所有限制性和修饰酶均购自New England Biolabs(NEB,Massachusetts,USA)。二元载体pEX2是用于利用潮霉素B显性选择的pPZP200衍生物[22]。
表1寡核苷酸的序列
Figure BDA0001286231190000231
Figure BDA0001286231190000241
Figure BDA0001286231190000251
以前已描述了各种启动子，如玉米黑粉菌(U.maydis)gpd1(Pgpd，长度为595bp)[31,32]、构巢曲霉(Aspergillus nidulans)gpdA(PgpdA,884bp)[33]、棉阿舒囊霉(Ashbyagossypii)翻译延伸因子1α基因(Ptef,348bp)[34]和RtGPD1(1429bp)[22]。利用质粒DNA作为模板并且分别利用Pgpd、PgpdA、Ptef和PRtGPD1的引物对Pgpd-Sf/Pgpd-Nr、PgpdA-Sf/PgpdA-Nr、Ptef-Sf/Ptef-Nr和Rt011S/Rt012N通过PCR获得启动子DNA片段。将所得的PCR片段用SpeI和NcoI消化并单独用于与1030bp BspHI/SmaI消化的合成hpt-3片段[22]和8855bp SpeI/SacI(平端)消化的载体pEC3GPD-GUS(图1)连接，以便分别产生pEC3GPD-HPT3、pEC3GPDA-HPT3、pEC3TEF-HPT3和pEC3GPDR-HPT3(图1)。
为了产生ALD1和DGA1的敲除突变体，利用圆红冬孢酵母ATCC 10657的总DNA作为模板并且分别利用寡核苷酸对ALD1Lf/ALD1Rr和Rt113/Rt114作为引物扩增完整或部分编码序列(分别为ALD1和DGA1的3kb和2.8kb)。在dNTP的存在下用T4DNA聚合酶处理之后将平端PCR产物连接至PmeI/SacI双消化的pEX2载体以产生中间质粒pEX2ALD1和pEX2DGA1，将扩增自质粒pRH2031的平端潮霉素抗性盒PGPD1::hpt-3::Tnos分别插入中间质粒pEX2ALD1和pEX2DGA1的XhoI/BspHI和SmaI/SpeI位点以产生基因靶向载体pKOALD1和pKODGA1。
分别利用圆红冬孢酵母ATCC 10657和R.glutinis ATCC 204091的cDNA模板扩增二酰甘油酰基-转移酶基因DGA1(GenBank登录号AB453835)和线粒体苹果酸酶基因MAE1(粘红酵母ATCC 204091基因组支架GL989657中的基因座标签RTG_03106)。引物对Rt055N/Rt056Ev和Rt057N/Rt058Ev分别用于DGA1和MAE1的扩增。将PCR产物用NcoI和EcoRV消化，与NcoI/EcoRV双消化的pRH2034连接，pRH2034包含蛋白表达盒，所述蛋白表达盒包含795bpRtGPD1启动子和花椰菜花叶病毒35S基因终止子和Cre-重组酶可切割的Umgpd::HPT-3:nos潮霉素选择盒[22]以产生pRHDGA1和pRHMAE1(图1)。
为了α-亚麻酸中的工程化研究，根据圆红冬孢酵母的密码子偏好合成密码子优化的编码高山被孢霉(Mortierella alpine)Δ12去饱和酶MaFAD2(SEQ ID NO:5)、亚麻(Linum usitatissimum)ω-3去饱和酶LuFAD3(GenBank登录号ABA02173.1；SEQ ID NO:10)和油桐(Vernicia fordii)(也称作Aleurites fordii)ω-3去饱和酶VfFAD3(SEQ ID NO:12)的基因，产生合成基因MaFAD2-2(SEQ ID NO:4)、LuFAD3-2(SEQ ID NO:9)和VfFAD3-2(SEQ ID NO:11)，将它们分别插入RtGPD1启动子调节下的pRH2034以产生pRHE001、pRHE002和pRHE003(图2)。为了产生FAD2-FAD3双基因过量表达盒，利用oligo Rt012Sf/35T-Pmr分别从质粒pRHE002和pRHE003扩增VfFAD3-2和LuFAD3-2盒，用SpeI(平端)和PmeI消化并与PmeI-cut pRHE001连接以产生质粒pRHE004和pRHE005(图3)。相似地，从pRHMAE1扩增MAE1盒，消化(SpeI-PmeI)，平端化，随后与PmeI-cut pRHDGA1连接以产生质粒pRHE006(图3)。
实施例3
根癌农杆菌(Agrobacterium tumefaciens)介导的转化
将二元载体电穿孔入根癌农杆菌AGL1(2.5kV,25μF,400Ω)，随后用补充了链霉素(100μg/ml)的2YT琼脂培养基进行选择。除非另有说明，如以前所述进行通过ATMT的真菌转化[22]。
实施例4
DNA印迹分析
如以前所述提取圆红冬孢酵母的基因组DNA[22]。将基因组DNA用PstI消化并通过在0.8％琼脂糖凝胶上电泳来分离，并且利用寡核苷酸HptRU和HptRSL2扩增DIG标记的部分hpt-3基因片段(nt 375-1036)的探针。为了基因缺失分析，将推定的敲除突变体Δald1和Δdga1的基因组DNA分别用HincII、PstI和HincII消化。分别利用oligo Rt148/Rt149和Rt113/Dga1-1扩增ALD1和DGA1的约0.6kb上游侧翼序列的DIG标记的探针。根据制造商的说明书(DIG-High prime DNA标记和检测启动试剂盒I，Roche Diagnostics)进行Southern杂交。
实施例5
定量逆转录PCR(q-RT-PCR)
如以前所述提取圆红冬孢酵母的总RNA[22]。为了去除污染DNA的痕迹，将RNA样品用DNase I(Roche Diagnostics,USA)处理，然后用乙醇沉淀。利用Improm-II逆转录系统(Promega,USA)合成cDNA，并且利用Platinum SYBR-Green qPCR SuperMix(Invitrogen,USA)在iCyclerTM实时PCR机器(Bio-Rad,USA)中进行实时PCR。实时条件如下：初始95℃变性步骤2min，然后在95℃下变性15s、在58℃下退火15s和在72℃下15s的延伸步骤的35个循环。利用iCyclerTM软件(Bio-Rad)获得数据。RtGPD1mRNA的表达水平用作靶基因表达归一化的参考。
实施例6
T-DNA标记位置的鉴定
利用高效热不对称InterLaced PCR(hiTAIL-PCR)鉴定基因组中的T-DNA标记位置[35,36]。特异性引物(HRSP1、HRSP2和HRSP3)和随机引物LAD1-4用于T-DNA左边界(LB)侧翼序列，而特异性引物(HRRSP1、HRRSP2和HRRSP3)和随机引物LAD1-4用于右边界(RB)侧翼序列。在PTC-200TM可编程热控制器(Bio-Rad,USA)中用i-Taq DNA聚合酶(i-DNA,Singapore)进行PCR反应。利用凝胶提取试剂盒(Qiagen)纯化PCR产物，并且利用BigDye终端试剂盒(Applied Biosystems,USA)用oligo HRRSP3(用于RB)或HRSP3(LB)直接测序。在某些情况下，将PCR产物克隆在pGTM-T easy载体(Promega,USA)中，并且利用oligo M13FP和M13RP作为引物测序。
实施例7
脂质积累突变体的筛选
通过随机插入pRH201的T-DNA将圆红冬孢酵母ATCC 90781基因组诱变。在补充了300μg/ml头孢噻肟、150μg/ml潮霉素以及50μg/ml浅蓝菌素、10μg/ml四氮唑紫或0.5μg/ml尼罗红(Sigma,USA)的YPD琼脂培养基上选择转化体。在28℃下温育5天之后，将表现出较大大小(针对浅蓝菌素选择)、较深紫色色素沉着(针对四氮唑紫选择)或较高荧光强度(在尼罗红中选择)的转化体转移至液体YPD培养基(300μg/ml头孢噻肟、150μg/ml潮霉素)用于增殖。在补充了上述抗生素的PDA平板上划线之后，将单克隆用于利用50ml培养物二次筛选以验证预期的表型。
实施例8
通过尼罗红染色比较脂质积累水平
如以前所述进行脂质含量快速估计的尼罗红染色[37]，有一些修改。简单地说，将10μl细胞培养物和2μl尼罗红储备溶液(丙酮中50mM)与200μl PBS缓冲液(pH7.4)在FluoroNunc平板(Thermo Fisher Scientific,Langenselbold,Germany)的孔中混合。每个样品伴有不含尼罗红的孔作为背景对照。将另一部分的细胞培养物(10μl)装载至96-孔平底透明平板(Nunc,Roskilde,Denmark)中的90μl PBS缓冲液(pH7.4)以测量细胞光密度。利用Infinite M200酶标仪(Tecan,Salzburg,Austria)利用iControlTM 3.0版本软件(Tecan,Salzburg,Austria)获得并分析数据。在推断背景对照之后在600nm下读取细胞光密度，同时分别用488nm和508nm的激发和发射波长测量荧光强度。通过在减去背景对照之后针对600nm下的吸收比归一化来计算相对脂质含量。在所有测试中，包括生物重复和统计学重复一式三份。
实施例9
通过GCMS的脂肪酸分析
如以前所述提取总脂质[38]，有一些修改。将细胞培养物(1ml)沉淀并用500μl脂质提取溶剂(氯仿:甲醇＝2:1)重悬。添加100μg玻璃珠(直径1mm，Sigma-Aldrich,Missouri,USA)之后，向混合物施加剧烈涡旋10min并用移液器去除溶剂相。如以前所述进行脂肪酸甲基酯(FAME)的制备和气液色谱(GC)分析[39]，有一些修改。将脂质旋转蒸发至几乎干燥(Concentrator,Eppendorf.USA)，溶于具有5％(vol/vol)H2SO4的1ml甲醇中，并且在密封的玻璃瓶中于90℃下温育2hr。添加1ml水中的PBS之后用300μl正己烷提取脂肪酸甲基酯。将1μl的己烷提取物注射至气相色谱质谱(GCMS QP2010,Shimadzu,Japan)中的DB-WAX熔融石英毛细管柱(30-m长度、0.25-μm直径和0.25-mm膜厚度)(Agilent J&WScientific,Folsom,CA,USA)上。运行条件通常为42.3ml/min氮流，起始温度180℃(3min)，15-min渐变至240℃，并且维持在240℃下7min。通过对Shimadzu NIST08化合物文库搜索鉴定脂肪酸甲基酯峰，并且定量为总脂肪酸的百分比(％TFA)。
实施例10
通过直接筛选圆红冬孢酵母T-DNA插入文库鉴定ALD1
已知T-DNA主要作为单拷贝整合入核基因组，并且这个特征已作为植物和真菌中的诱变工具广泛利用[32,40-43]。为了研究是否可以通过直接筛选T-DNA突变体文库鉴定调节油收率或质量的新基因，我们设计了三个独立的筛选策略，旨在利用药物或荧光染料的辅助鉴定T-DNA突变体中脂肪酸谱或含量的变化。
浅蓝菌素(2S)(3R)2,3-环氧基-4-氧代-7,10-十二碳二烯酰胺是分离自Cephalosporium caerulens的培养肉汤的药物[44,45]，并且已成功用于提高细胞内多不饱和脂肪酸的积累[46]或产油微生物中的脂质含量[47,48]。由于其阻断脂肪酸生物合成的能力用作杀真菌剂，预期在这个处理中存活的突变体具有较高水平的脂质或多不饱和脂肪酸。我们针对YPD琼脂培养基中50μg/ml浅蓝菌素筛选了～10,000个转化体，并且发现12个突变体看来对浅蓝菌素更耐受。我们将这些推定的红冬孢酵母属浅蓝菌素突变体分别命名为RCM1至RCM12。虽然发现在小规模液体培养物中脂质含量与Wt有点不同，但是RCM表现出显著较高水平的α-亚麻酸(ALA)(图4)。值得注意的是，RCM6产生比WT高3-倍以上水平的ALA(图4B)。
其次，尼罗红(NR)已广泛用作脂质的荧光示踪剂[50]。通过筛选～10,000个T-DNA突变体，我们鉴定了看来表现出较强红色荧光的4个候选，将它们命名为RNM1-4(用于红冬孢酵母属尼罗红突变体)。脂质收率的定量显示与WT相比在RNM突变体中显著提高(图4D)。但是，可以观察到脂肪酸组成差异很小(数据未示出)。
相似地，将用作脂质积累的染料指示剂[51]或微生物生长的氧化还原指示剂[52]的四氮唑紫用作指示剂以筛选～3,000个转化体，导致鉴定了6个较深着色的突变体(图4C)。但是，脂肪酸谱的重复分析不能验证脂肪酸积累的改变(数据未示出)。
通过Hi-TAIL PCR技术鉴定上述突变体中的T-DNA标记位置，并且结果显示成功获得并测序了12个RCM中的11个、4个RNM中的2个和6个RTM中的6个。受到影响的基因主要参与细胞壁整合的维持，脂质代谢，信号转导，蛋白折叠和运输，次级代谢物、氨基酸、维生素、辅因子等的代谢(表2)。
表2
圆红冬孢酵母浅蓝菌素突变体(RCM)、圆红冬孢酵母四唑紫突变体(RTM)、圆红冬孢酵母尼罗红突变体(RNM)和圆红冬孢酵母白化突变体(RAM)的T-DNA标记位置
Figure BDA0001286231190000301
Figure BDA0001286231190000311
Figure BDA0001286231190000321
aLB-侧翼序列
b根据BLASTx结果确定T-DNA标记的基因
c上游1.0kb、上游0.5kb和下游0.3kb分别表示相应标记基因的上游501～1000bp、500bp和下游300bp内的T-DNA插入
d最佳命中表示具有最高E-评分的BLASTx结果
e根据BLASTx结果确定注释
f微生物表示最佳命中的宿主
g相同性值来自BLASTx结果
h由于坏的测序结果不可用
实施例11
T-DNA标记突变体RCM6的表征
为了进一步研究通过上述正向遗传学筛选的突变体，并且作为原理验证，进行反向遗传学分别用于突变体RCM6和RAM5中对脂质积累和类胡萝卜素生物合成的潜在调节作用的研究。
同源分析显示RCM6中的T-DNA整合在400th重叠群(contig)的72542-72543nt内(GenBank登录号AEVR01000400)。T-DNA标记位置相邻序列的BLASTx显示推定的包含醛脱氢酶结构域的蛋白被RCM6中的T-DNA整合破坏(图6A)。靶蛋白位于推定的噻唑生物合成酶(EGU11956)和候选二肽基氨基肽酶(EGU11957)之间，显示与来自其他真菌物种如Streptomyces sviceus(EDY60340.1，E-值＝2E-68)和分枝杆菌属(Mycobacterium sp.)(YP_936108，E-值＝3E-66)等的醛氢化酶高度同源。因此，被T-DNA破坏的推定的醛氢化酶(命名为ALD1)编码基因会导致RCM6的表型。RT-PCR和快速扩增cDNA末端(RACE)的进一步分析显示ALD1基因跨越重叠群#400中的2461bp，包含10内含子分开的11个外显子(图5A)。其5’UTR长度为18nt，然后是一段短的富含CT的基序(CT盒，数据未示出)。mRNA剪接严格遵守经典的GU-AG规则，其产生长度为1506-nt的mRNA，编码501-aa蛋白，具有严格保守的NAD-结合指纹基序Gly-X-Gly-X-X-Gly(SEQ ID NO:53)(GSGTVG,aa 193-198；SEQ ID NO:54)、腺苷核糖(NAD)-结合氨基酸(E148)和催化中心Cys(C249)(图6)。在RCM6中，T-DNA整合入从ALD1的起始密码子第2097和第2098核苷酸之间的位置，破坏第9外显子，导致缺失形成部分RYPP基序的C-端58aa(图6)。
为了进一步证实在脂质积累中的功能，通过同源重组在ATMT的辅助下缺失ALD1。用潮霉素抗性盒(PGPD1::hpt-3::Tnos，图5A)置换范围为+536至+1947的ALD1中的核苷酸序列，并且通过DNA印迹分析验证正确的ald1空突变体(图5B)。当在液体肉汤中或在琼脂培养基上培养时，与Wt中粉色着色的颜色相比，Δald1表现出橙色着色的颜色(图5E)。Δald1在脂质积累培养基中生长比WT略慢，但是当培养基中的碳源在第4天耗尽时其产生相似的生物质(图5E)。在葡萄糖耗尽之前(第3天)，Δald1突变体在脂质收率中表现与WT差异很少。当葡萄糖耗尽时，脂质水平在两个菌株中均降低，但是，脂质含量在Δald1突变体中显著较高(图5C和5D)。惊人的是，几乎一半的ALA在WT中降解，而在Δald1突变体中发现很少降解(图5F)。这些结果与T-DNA标记突变体RCM6的结果一致(图4B)。
实施例12
通过合理设计提高圆红冬孢酵母中的总TAG积累
三酰甘油(TAG)是大多数真核细胞中存在的主要中性脂质，并且生物合成途径是高度保守的[53]。通过合理设计的代谢工程在提高脂质含量和生产率中已非常成功[54-56]。在圆红冬孢酵母中，可获得二酰甘油乙酰转移酶(Dga1)和苹果酸酶(MAE1)的序列信息(分别参见例如SEQ ID NO:81和SEQ ID NO:84)。构建两个基因的过量表达盒，通过795-bpRtGPD1启动子PGPD1::DGA1和PGPD1::MAE1驱动，图1D，通过根癌农杆菌介导的转化将其整合入圆红冬孢酵母ATCC 10657的染色体。定量RT-PCR分析显示两个基因的mRNA转录物水平在3-天生物过程中显著增强(图7A)，导致DGA1和MAE1过量表达菌株比Wt菌株分别提高2.3和1.8-倍的峰脂质收率(图7B)。在两个菌株中脂肪酸谱未显著改变(图7C和7F)。正如预期的，构建的空突变体dga1(图7D)具有显著减少的脂质积累(图5E)。
实施例13
通过基本原理设计提高圆红冬孢酵母中的多不饱和脂肪酸(PUFA)生成
在圆红冬孢酵母菌株中，油酸(C18:1)是主要脂肪酸组分(～50％)，而棕榈油酸(C16:1)和亚油酸(C18:2)组成～20％的总FA。多不饱和ω-3脂肪酸α-亚麻酸(C18:3n＝9,ALA)是以总脂肪酸的3～4％(％TFA)存在的微量组分(图8)。为了从油酸产生ALA，需要δ-12去饱和酶(Fad2)和ω-3去饱和酶(Fad3)[57,58]。根据圆红冬孢酵母的密码子使用偏好设计并商业合成亚麻(L.usitatissimum)FAD3、桐树(油桐(V.fordii))FAD3和高山被孢霉(M.alpine)FAD2[22]，其中用圆红冬孢酵母中常用的密码子代替所有稀有密码子。涉及的基因的GC含量为LuFAD3-2(SEQ ID NO:9)中的65.3％，MaFAD2-2(SEQ ID NO:4)中的64.8％和VfFAD3-2(SEQ ID NO:11)中的63.3％。
通过可操作地连接至RtGPD1启动子和花椰菜花叶病毒35S终止子实现3个合成基因的高水平表达，然后分析所选ATMT菌株的脂肪酸组成。过量表达LuFAD3-2(RHE001)、VfFAD3-2(RHE002)或MaFAD2-3(RHE003)的优良菌株表现出ALA含量分别提高1.8、2.2和1.6倍(图9A)。随后，将RtGPD1::AfFAD3-2:35S和RtGPD1::MaFAD2-2:35S安插入单个T-DNA载体pRHE004，通过ATMT将其转化至圆红冬孢酵母ATCC 10657的衍生物(RT1CE6，包含稳定整合入基因组的17β-雌二醇诱导Cre基因)。通过利用pKOALD1载体缺失ALD1基因来进一步修饰所选优良菌株(RHE004)。所得菌株(命名为ald1eAM)产生比ald1空突变体(ald1e)高3.74-倍的ALA(图9B)，ALA含量达到总脂肪酸的～49％。
实施例14
Ald1的生物化学分析
为了表征Ald1并证实RCM6突变体中的T-DNA插入所致的C-端58个残基缺失损害其酶活性，将Ald1蛋白的全长和截短版本在大肠杆菌(E coli)BL21(DE3)中作为具有C-端6X组氨酸标签的融合蛋白表达。将重组Ald1和Ald1n用HisTrap柱(GE healthcare,USA)纯化并利用以前报道的方法测定[59]，有一些修改。简单地说，反应混合物由40μl的100mMTris-Cl缓冲液(pH8.0)、30μl的10mM NAD+或NADP+(Sigma-Aldrich,USA)、10μl的20mM十二醛(十二烷基醛，C12-醛，Sigma-Aldrich,USA)、110μl水和10μl纯化的酶组成。反应在室温(25℃)下进行，并且通过添加酶起始。如以前所述[40]，利用iControlTM版本3.0软件(Tecan,Salzburg,Austria)通过Infinite M200酶标仪(Tecan,Salzburg,Austria)读取340nm下光密度值的时间进程。如图10所示，Ald1和Ald1n均表现出明显的脱氢酶活性，略微偏好NAD+。值得注意的是，具有C-端58aa缺失的突变蛋白表现出显著较低的酶促活性。
实施例15
圆红冬孢酵母中脂肪酸生物合成基因的表征
通过利用已知的解脂耶氏酵母和玉米黑粉菌酶序列作为询问对圆红冬孢酵母ATCC 204091基因组支架序列204091(以前名为粘红酵母，GenBank登录号AEVR02000000，全基因组鸟枪测序计划PRJNA59971，密西西比州立大学，USA.)进行BLAST搜索鉴定了各种脂肪酸去饱和酶、延长酶和ATP-柠檬酸裂解酶的圆红冬孢酵母同源物。用圆红冬孢酵母菌株ATCC 10657或其衍生物Rt1ck(在同源重组中表现出极高效率的KU70缺陷突变体)进行遗传操纵和DNA序列表征。使用的寡核苷酸在表3中列出。
表3用于以下的寡核苷酸：
用于硬脂酰-CoA-δ-9-去饱和酶基因(RtFAD1)的缺失
Figure BDA0001286231190000351
用于δ-12去饱和酶基因(RtFAD2)的缺失
Figure BDA0001286231190000352
Figure BDA0001286231190000361
用于延长酶基因1(RtELO1)的缺失
Figure BDA0001286231190000362
用于ATP-柠檬酸裂解酶基因(RtACL1)的缺失
Figure BDA0001286231190000363
用于延长酶基因2(RtELO2)的缺失
Figure BDA0001286231190000364
用于ELO1 cDNA的表达
Rt236Nf TTTCCATGGCCTCGTACGCCGCCCATCC(143) NcoI
Rt237Evr TTTGATATCGGAAGGGACCGCGCTAGTTG(144) EcoRV
用于ELO2 cDNA的表达
Rt259Nf TTTCCATGGTCGCACCGTCCCCCG(145) NcoI
Rt260Evr TTTGATATCGAGAAGAAGGTGGGGTGTTTAG(146) EcoRV
________________
aSEQ ID NO:
为了δ-9-油酸去饱和酶基因FAD1(或OLE1)同源物的缺失，利用圆红冬孢酵母ATCC10657基因组DNA用寡核苷酸对DS9L-Sf/DS9L-Br和DS9R-Hf/DS9R-Str分别扩增左侧和右侧同源侧翼片段(各～0.9kb)。用SacI/PmeI消化的pEX2二元载体、SacI/BamHI消化的左侧侧翼片段、BamHI/HindIII消化的来自pDXP795hptR的密码子优化的潮霉素选择盒(PGPD1::hpt-3::Tnos,[60])和HindIII/StuI消化的右侧侧翼片段进行4-片段连接以产生基因缺失质粒pKOOLE1。应用相似的策略构建pKOFAD2和pKOELO1，分别用于敲除推定的δ-12去饱和酶基因和延长酶基因1。Oligo对DS12L-Sf2/DS12L-Br2和DS12R-Hf/DS12R-Str用来扩增pKOFAD2的左侧(0.6kb)和右侧(0.9kb)同源侧翼片段，并且ELO1L-Sf/ELO1L-Br和ELO1R-Hf/ELO1R-Str用于pKOELO1(各～0.9kb)。对于ELO2，对于左侧和右侧同源侧翼片段，分别使用oligo对ELO2L-Stf/ELO2L-Hr和ELO2R-Bf/ELO2R-Sr(各～0.8kb)并用StuI/HindIII和BamHI/SacI消化。在4-片段连接中相似地使用两个片段以产生pKOELO2。
为了缺失推定的ATP-柠檬酸裂解酶基因(RtACL1)，将寡核苷酸o对ACL1L-Sf2/ACL1L-Br2和ACL1R-Hf2/ACL1R-Str2用来扩增左侧和右侧同源侧翼片段(各0.9kb)以利用上文所述相似的策略产生pKOACL1。
通过RT-PCR获得所关注的基因的cDNA序列，利用BD SMARTerTM RACE cDNA扩增试剂盒(Clontech,California,USA)根据制造商的说明书进行5’和3’RACE。寡核苷酸对OLE1U1/OLE1L1、FAD2U1/FAD2L1分别用作FAD1(OLE1)和FAD2的5’/3’RACE的特异性引物。
预测的Fad1(Ole1)和Fad2的ORF分别编码545、451aa的蛋白。两个Fad享有膜脂肪酸去饱和酶的共有保守结构域(蛋白家族编号pfam00487,EMBL-EBI)。但是，Fad2缺少细胞色素β5样血红素/类固醇结合结构域(pfam00173。BLAST搜索显示Ole1和Fad2分别与来自的禾柄锈菌(Puccinia graminis)的硬脂酰-CoA去饱和酶(XP_003326562.1，70％相同性)、来自玉米黑粉菌的Δ12-脂肪酸去饱和酶(XP_757193.1，57％相同性)表现出最高相同性。
鉴定了两种延长酶。ELO1(Seq ID.No.99和100)和ELO2(Seq ID.No.102和103)分别编码长度为329(Seq ID 101)和293aa(Seq ID 104)的蛋白。两种推定的脂肪酸延长酶均享有参与长链脂肪酸延长系统的GNS1/SUR4家族的共有保守结构域(pfam01151)。Elo1和Elo2分别与来自禾柄锈菌(PGTG06945，XP_003325743.2，43％相同性)和松杨栅锈菌(MELLADRAFT_42723，XP_007407925.1，65％相同性)的脂肪酸延长酶表现出最高相同性。ELO1和ELO2敲除的脂肪酸谱分析显示ELO1敲除导致谱变化很少，除了C18:0适度减少以及C16:0和C18:1少量增加。相比之下，ELO2敲除导致长链脂肪酸(>C18)合成的完全丧失(图12)。这些结果强烈表明OLE1是短链脂肪-CoA延长酶，而ELO2能够是长链和短链脂肪酸-CoA的延长酶。
为了过量表达研究，利用通过逆转录合成的圆红冬孢酵母cDNA分别用引物对Rt227Nf/Rt228Evr、Rt229Ndf/Rt230Evr和Rt259Nf/Rt260Evr扩增OLE1、FAD2和ELO2的cDNA。将NcoI/EcoRV消化的PCR产物与NcoI/EcoRV消化的pKC1载体连接以产生pKC1OLE1、pKC1FAD2和pKC1ELO2，因为载体pKC1中使用的强RtGPD1启动子导致基因的过量表达。
实施例16
基因缺失分析
为了证实每个基因的功能，通过各敲除构建体的农杆菌(Agrobacterium)介导的转化产生敲除突变体；通过菌落PCR和DNA印迹分析筛选。FAD1敲除在几次尝试中不成功。FAD2的缺失在转化和增殖培养基中补充亚油酸之后成功。亚油酸(C18:2,LA)和α-亚麻酸(C18:3,ALA)在FAD2空突变体中不存在，而C18:1的含量增加至总脂肪酸的几乎70％(图11B)。这证实FAD2基因(SEQ ID NO:92)编码催化油酸(C18:1)转化形成LA的Δ12-脂肪酸去饱和酶。FAD2敲除突变体中缺少ALA表明Fad2是Δ12和Δ15双功能脂肪酸去饱和酶。甚至在补充了LA(C18:2)前体时fad2Δ突变体不能产生ALA生成支持这个(图11D)。
通过过量表达可以证实FAD1(OLE1)的功能。将RtGDP1::OLE1盒(图13A)转化入Wt和ALD1敲除菌株(Δald1e)导致油酸含量显著增加(图14)。因此，将RtGDP1::OLE1叠加至MaFAD2-2(图13D)，或者叠加至MaFAD2-2和VfFAD3-2过量表达盒(图13E)分别导致LA和ALA增加(图14)。在表达3-基因盒的18个转化体中，在ald1e背景中3个表现出>20％ALA含量，一个包含～24％。这个系列中较低的ALA含量可能归因于用于MaFAD2-2的较弱ACC1启动子，因为LA含量非常低(未示出)。
实施例17
在圆红冬孢酵母中表征ATP-柠檬酸裂解酶(ACL1)基因
基于动物中的研究，据信真菌ATP-柠檬酸裂解酶(ACL)是油积累的重要因子。通过粘红酵母ATCC 204091部分基因组序列的BLAST搜索鉴定了推定的ATP-柠檬酸裂解酶/合成酶基因ACL1(seq ID No.86和87)。推定的Acl1蛋白序列在Seq ID.No.88中示出。产生ACL1基因的敲除突变体，并且其表现出显著减少的油积累和生物质生长(图15)。这强烈表明ACL1表达促进圆红冬孢酵母中的油积累和生物质生成。
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序列表
<110> 淡马锡生命科学研究院有限公司
<120> 在红冬孢酵母属和红酵母属物种中高效产生多不饱和脂肪酸(PUFA)的方法
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<150> US 60/047,300
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ctcgctcgct tgcgtgtcgt cctcgctttc ggagcttcga aacaggcggg acgaggggat 120
cgtggtgcag cgcgcgcctg gggaggattc gctggacatc ggggatattg ctggaggata 180
ctcaggactt tctggattgg tcccgccctt cccgcgcccg tcgtcgctct ccagcatcca 240
ttccccactc gacactccgg caaactcgcc gctgactctc cccttcgctc attccgcagg 300
cttacgacac cgtcaccaag gcgttcttgt cgggaaagac taggcccatc gcctggcgca 360
aggcgcagat caaaaagctc gggttccttg tcgtgagttc gcggttcttt cgcgcggtct 420
ttgcgacggc tcgtccagtg caggtcgcgc tggtgcgcgt cgtcaatgta gttgcgatga 480
aacctgaccc agcaactccc cctcccacag caagacaacg aagacgcctt cgtccgcgct 540
cttgagcaag actttggccg cccagccttc gagacaatca ccgccgagat caaccccgtc 600
aaggctgaga tcaacgaggt ctacgaccac ctcgagaagt gggccaagcc gaggcgcgtc 660
aagacttcgg cgacgtggta cgctaccaag ccgacggtct actcggagcc taagggtgtc 720
acgctcgtta tcgggacgtg gaactgtgcg tgcacttgtt ttgtacgagg ttgagtgtgt 780
gtactgacgg aagatgtcgc cgcagacccg atcacgctcc tcctcgtccc gcttctcggc 840
gccatctctg ccggctgcac cgcgctcgtc aaggtgcctt gccgttcgaa gtccgtcgta 900
ctgcatgtct cactgacact cgtcgctccc acagcccgct gagcaagccc ctcacgtcgc 960
cgcgctcgtc gccgacctcc tgcccaagta cctcgacccc accgccttca tctgcatcaa 1020
cggcgccatc cctcaagcga ccgctctcct caaactcaag ttcgatcaca tcttctacac 1080
cggttcggga acggtcggca agatcgttgc gcgtgcggcg gcggagcacc tttgcccggt 1140
tacgcttgag ttggggggaa agagtccggc ggttgtgctg gatgatgcgg atattgaggt 1200
tgtggcgagg aggatcgttt gggccaagtt taccaacgct gggcaggtgc gtcgcgagaa 1260
ccgggttgtg tcgttggtct cgccgaagcg ggcgcagatg cttagtcatc cgcttgttgt 1320
tgcgcacaga tctgcatctc gacagactac gtcctcacga ccccgcagac cgagcccaag 1380
ctcctcgaag ccctcaagcg cgctctcgcc gccttctccg ccaaccccgc cgcctcctcc 1440
tcctcggaaa agtcgtcaac ctcgctcgtg cacaacccga actactcgcg catcatcaac 1500
cagaaccact acaaccgcgt ttcgaagttg cttgatgcga ctaagggcga ggtggttgtt 1560
ggcggcggga gggacgagaa ggagcgcaag atcgaggtca cgatcgtgag gggcgttaag 1620
ccggatgact cgctcatgtc gggtgcgcac tgcggctctc cccctgaaga acgaatgtgg 1680
ctgacgaatg cgaccgagca gaggagattt tcggccccgt ccccccgatc gtgaccctcc 1740
cgacgctcga cgacatggtc aagttcattc agtcgcgcga cacgcctctc gcgctttacg 1800
tcttcacgca gagcaggaag aaccgcgact ttagtgcgtt ccccgcctct ctctcgctcg 1860
ctgaccttcg actgactcgg tggtggatgc agtttttgag cgcactcgct cgggcggatt 1920
cgttcagaac gatgtgctcg ttcagttcat gatccctggg ttgccgttcg gcggtacggg 1980
cgcggcgggc tacggaaact accacggcag gcggtgcgtc ccgcttcttc ggcaccgtgc 2040
ttccccgagt ccggctgacc tgctcgcacg cagcaccttc gacacgttct cgcacgagcg 2100
cgcgtcggcc aatgtcccca cctggatgga catgatcatg gcgtcgcggt accctcccta 2160
cacccgttcg tccgggtcgt cccttcctct ctgtgctcgc taacacactc gtcccacacg 2220
tgcagagaag aagctgaaga tgctcctgtt cgcgaccaag gcggtgatca agaagcccag 2280
caagtttggc tcgatctcgc gcttgctcaa ggtgattgcc gcgatggtcg ctctcttggc 2340
tgtcagggcc aggctctgac tgacccgtcg tcgtcccacc cctccccttc tccaccactc 2400
ctcttcctct cgggtcttgg gaatcgtgtg cgctgggcag aagttgacgg gacaggcgtg 2460
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Met Ala Ala Met Gln Asp Thr Pro Ile Asp Ser Ile Pro Gln Ala Tyr
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gac acc gtc acc aag gcg ttc ttg tcg gga aag act agg ccc atc gcc 96
Asp Thr Val Thr Lys Ala Phe Leu Ser Gly Lys Thr Arg Pro Ile Ala
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tgg cgc aag gcg cag atc aaa aag ctc ggg ttc ctt gtc caa gac aac 144
Trp Arg Lys Ala Gln Ile Lys Lys Leu Gly Phe Leu Val Gln Asp Asn
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gaa gac gcc ttc gtc cgc gct ctt gag caa gac ttt ggc cgc cca gcc 192
Glu Asp Ala Phe Val Arg Ala Leu Glu Gln Asp Phe Gly Arg Pro Ala
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Phe Glu Thr Ile Thr Ala Glu Ile Asn Pro Val Lys Ala Glu Ile Asn
65 70 75 80
gag gtc tac gac cac ctc gag aag tgg gcc aag ccg agg cgc gtc aag 288
Glu Val Tyr Asp His Leu Glu Lys Trp Ala Lys Pro Arg Arg Val Lys
85 90 95
act tcg gcg acg tgg tac gct acc aag ccg acg gtc tac tcg gag cct 336
Thr Ser Ala Thr Trp Tyr Ala Thr Lys Pro Thr Val Tyr Ser Glu Pro
100 105 110
aag ggt gtc acg ctc gtt atc ggg acg tgg aac tac ccg atc acg ctc 384
Lys Gly Val Thr Leu Val Ile Gly Thr Trp Asn Tyr Pro Ile Thr Leu
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ctc ctc gtc ccg ctt ctc ggc gcc atc tct gcc ggc tgc acc gcg ctc 432
Leu Leu Val Pro Leu Leu Gly Ala Ile Ser Ala Gly Cys Thr Ala Leu
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Val Lys Pro Ala Glu Gln Ala Pro His Val Ala Ala Leu Val Ala Asp
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Leu Leu Pro Lys Tyr Leu Asp Pro Thr Ala Phe Ile Cys Ile Asn Gly
165 170 175
gcc atc cct caa gcg acc gct ctc ctc aaa ctc aag ttc gat cac atc 576
Ala Ile Pro Gln Ala Thr Ala Leu Leu Lys Leu Lys Phe Asp His Ile
180 185 190
ttc tac acc ggt tcg gga acg gtc ggc aag atc gtt gcg cgt gcg gcg 624
Phe Tyr Thr Gly Ser Gly Thr Val Gly Lys Ile Val Ala Arg Ala Ala
195 200 205
gcg gag cac ctt tgc ccg gtt acg ctt gag ttg ggg gga aag agt ccg 672
Ala Glu His Leu Cys Pro Val Thr Leu Glu Leu Gly Gly Lys Ser Pro
210 215 220
gcg gtt gtg ctg gat gat gcg gat att gag gtt gtg gcg agg agg atc 720
Ala Val Val Leu Asp Asp Ala Asp Ile Glu Val Val Ala Arg Arg Ile
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Val Trp Ala Lys Phe Thr Asn Ala Gly Gln Ile Cys Ile Ser Thr Asp
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tac gtc ctc acg acc ccg cag acc gag ccc aag ctc ctc gaa gcc ctc 816
Tyr Val Leu Thr Thr Pro Gln Thr Glu Pro Lys Leu Leu Glu Ala Leu
260 265 270
aag cgc gct ctc gcc gcc ttc tcc gcc aac ccc gcc gcc tcc tcc tcc 864
Lys Arg Ala Leu Ala Ala Phe Ser Ala Asn Pro Ala Ala Ser Ser Ser
275 280 285
tcg gaa aag tcg tca acc tcg ctc gtg cac aac ccg aac tac tcg cgc 912
Ser Glu Lys Ser Ser Thr Ser Leu Val His Asn Pro Asn Tyr Ser Arg
290 295 300
atc atc aac cag aac cac tac aac cgc gtt tcg aag ttg ctt gat gcg 960
Ile Ile Asn Gln Asn His Tyr Asn Arg Val Ser Lys Leu Leu Asp Ala
305 310 315 320
act aag ggc gag gtg gtt gtt ggc ggc ggg agg gac gag aag gag cgc 1008
Thr Lys Gly Glu Val Val Val Gly Gly Gly Arg Asp Glu Lys Glu Arg
325 330 335
aag atc gag gtc acg atc gtg agg ggc gtt aag ccg gat gac tcg ctc 1056
Lys Ile Glu Val Thr Ile Val Arg Gly Val Lys Pro Asp Asp Ser Leu
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atg tcg gag gag att ttc ggc ccc gtc ccc ccg atc gtg acc ctc ccg 1104
Met Ser Glu Glu Ile Phe Gly Pro Val Pro Pro Ile Val Thr Leu Pro
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acg ctc gac gac atg gtc aag ttc att cag tcg cgc gac acg cct ctc 1152
Thr Leu Asp Asp Met Val Lys Phe Ile Gln Ser Arg Asp Thr Pro Leu
370 375 380
gcg ctt tac gtc ttc acg cag agc agg aag aac cgc gac ttt att ttt 1200
Ala Leu Tyr Val Phe Thr Gln Ser Arg Lys Asn Arg Asp Phe Ile Phe
385 390 395 400
gag cgc act cgc tcg ggc gga ttc gtt cag aac gat gtg ctc gtt cag 1248
Glu Arg Thr Arg Ser Gly Gly Phe Val Gln Asn Asp Val Leu Val Gln
405 410 415
ttc atg atc cct ggc ttg ccg ttc ggc ggt acg ggc gcg gcg ggc tac 1296
Phe Met Ile Pro Gly Leu Pro Phe Gly Gly Thr Gly Ala Ala Gly Tyr
420 425 430
gga aac tac cac ggc agg cgc acc ttc gac acg ttc tcg cac gag cgc 1344
Gly Asn Tyr His Gly Arg Arg Thr Phe Asp Thr Phe Ser His Glu Arg
435 440 445
gcg tcg gcc aat gtc ccc acc tgg atg gac atg atc atg gcg tcg cgg 1392
Ala Ser Ala Asn Val Pro Thr Trp Met Asp Met Ile Met Ala Ser Arg
450 455 460
tac cct ccc tac acc cag aag aag ctg aag atg ctc ctg ttc gcg acc 1440
Tyr Pro Pro Tyr Thr Gln Lys Lys Leu Lys Met Leu Leu Phe Ala Thr
465 470 475 480
aag gcg gtg atc aag aag ccc agc aag ttt ggc tcg atc tcg cgc ttg 1488
Lys Ala Val Ile Lys Lys Pro Ser Lys Phe Gly Ser Ile Ser Arg Leu
485 490 495
ctc aag aag ttg acg gga cag gcg tga 1515
Leu Lys Lys Leu Thr Gly Gln Ala
500
<210> 3
<211> 504
<212> PRT
<213> Rhodosporidium toruloides
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Met Ala Ala Met Gln Asp Thr Pro Ile Asp Ser Ile Pro Gln Ala Tyr
1 5 10 15
Asp Thr Val Thr Lys Ala Phe Leu Ser Gly Lys Thr Arg Pro Ile Ala
20 25 30
Trp Arg Lys Ala Gln Ile Lys Lys Leu Gly Phe Leu Val Gln Asp Asn
35 40 45
Glu Asp Ala Phe Val Arg Ala Leu Glu Gln Asp Phe Gly Arg Pro Ala
50 55 60
Phe Glu Thr Ile Thr Ala Glu Ile Asn Pro Val Lys Ala Glu Ile Asn
65 70 75 80
Glu Val Tyr Asp His Leu Glu Lys Trp Ala Lys Pro Arg Arg Val Lys
85 90 95
Thr Ser Ala Thr Trp Tyr Ala Thr Lys Pro Thr Val Tyr Ser Glu Pro
100 105 110
Lys Gly Val Thr Leu Val Ile Gly Thr Trp Asn Tyr Pro Ile Thr Leu
115 120 125
Leu Leu Val Pro Leu Leu Gly Ala Ile Ser Ala Gly Cys Thr Ala Leu
130 135 140
Val Lys Pro Ala Glu Gln Ala Pro His Val Ala Ala Leu Val Ala Asp
145 150 155 160
Leu Leu Pro Lys Tyr Leu Asp Pro Thr Ala Phe Ile Cys Ile Asn Gly
165 170 175
Ala Ile Pro Gln Ala Thr Ala Leu Leu Lys Leu Lys Phe Asp His Ile
180 185 190
Phe Tyr Thr Gly Ser Gly Thr Val Gly Lys Ile Val Ala Arg Ala Ala
195 200 205
Ala Glu His Leu Cys Pro Val Thr Leu Glu Leu Gly Gly Lys Ser Pro
210 215 220
Ala Val Val Leu Asp Asp Ala Asp Ile Glu Val Val Ala Arg Arg Ile
225 230 235 240
Val Trp Ala Lys Phe Thr Asn Ala Gly Gln Ile Cys Ile Ser Thr Asp
245 250 255
Tyr Val Leu Thr Thr Pro Gln Thr Glu Pro Lys Leu Leu Glu Ala Leu
260 265 270
Lys Arg Ala Leu Ala Ala Phe Ser Ala Asn Pro Ala Ala Ser Ser Ser
275 280 285
Ser Glu Lys Ser Ser Thr Ser Leu Val His Asn Pro Asn Tyr Ser Arg
290 295 300
Ile Ile Asn Gln Asn His Tyr Asn Arg Val Ser Lys Leu Leu Asp Ala
305 310 315 320
Thr Lys Gly Glu Val Val Val Gly Gly Gly Arg Asp Glu Lys Glu Arg
325 330 335
Lys Ile Glu Val Thr Ile Val Arg Gly Val Lys Pro Asp Asp Ser Leu
340 345 350
Met Ser Glu Glu Ile Phe Gly Pro Val Pro Pro Ile Val Thr Leu Pro
355 360 365
Thr Leu Asp Asp Met Val Lys Phe Ile Gln Ser Arg Asp Thr Pro Leu
370 375 380
Ala Leu Tyr Val Phe Thr Gln Ser Arg Lys Asn Arg Asp Phe Ile Phe
385 390 395 400
Glu Arg Thr Arg Ser Gly Gly Phe Val Gln Asn Asp Val Leu Val Gln
405 410 415
Phe Met Ile Pro Gly Leu Pro Phe Gly Gly Thr Gly Ala Ala Gly Tyr
420 425 430
Gly Asn Tyr His Gly Arg Arg Thr Phe Asp Thr Phe Ser His Glu Arg
435 440 445
Ala Ser Ala Asn Val Pro Thr Trp Met Asp Met Ile Met Ala Ser Arg
450 455 460
Tyr Pro Pro Tyr Thr Gln Lys Lys Leu Lys Met Leu Leu Phe Ala Thr
465 470 475 480
Lys Ala Val Ile Lys Lys Pro Ser Lys Phe Gly Ser Ile Ser Arg Leu
485 490 495
Leu Lys Lys Leu Thr Gly Gln Ala
500
<210> 4
<211> 1206
<212> DNA
<213> Mortierella alpine
<220>
<221> CDS
<222> (1)..(1206)
<400> 4
atg gcc ccg ccg aac acc atc gac gcc ggc ctc acc cag cgc cac atc 48
Met Ala Pro Pro Asn Thr Ile Asp Ala Gly Leu Thr Gln Arg His Ile
1 5 10 15
tcg acc tcg gcc gcc ccg aac tcg gcc aag ccg acc ttc gag cgc aac 96
Ser Thr Ser Ala Ala Pro Asn Ser Ala Lys Pro Thr Phe Glu Arg Asn
20 25 30
tac cag ctc ccg gag ttc acc atc aag gag atc cgc gag tgc atc ccg 144
Tyr Gln Leu Pro Glu Phe Thr Ile Lys Glu Ile Arg Glu Cys Ile Pro
35 40 45
gcc cac tgc ttc gag cgc tcg ggc ctc cgc ggc ctc tgc cac gtc gcc 192
Ala His Cys Phe Glu Arg Ser Gly Leu Arg Gly Leu Cys His Val Ala
50 55 60
atc gac ctc acc tgg gcc tcg ctc ctc ttc ctc gcc gcc acc cag atc 240
Ile Asp Leu Thr Trp Ala Ser Leu Leu Phe Leu Ala Ala Thr Gln Ile
65 70 75 80
gac aag ttc gag aac ccg ctc atc cgc tac ctc gcc tgg ccg gtc tac 288
Asp Lys Phe Glu Asn Pro Leu Ile Arg Tyr Leu Ala Trp Pro Val Tyr
85 90 95
tgg atc atg cag ggc atc gtc tgc acc ggc atc tgg gtc ctc gcc cac 336
Trp Ile Met Gln Gly Ile Val Cys Thr Gly Ile Trp Val Leu Ala His
100 105 110
gag tgc ggc cac cag tcg ttc tcg acc tcg aag acc ctc aac aac acc 384
Glu Cys Gly His Gln Ser Phe Ser Thr Ser Lys Thr Leu Asn Asn Thr
115 120 125
gtc ggc tgg atc ctc cac tcg atg ctc ctc gtc ccg tac cac tcg tgg 432
Val Gly Trp Ile Leu His Ser Met Leu Leu Val Pro Tyr His Ser Trp
130 135 140
cgc atc tcg cac tcg aag cac cac aag gcc acc ggc cac atg acc aag 480
Arg Ile Ser His Ser Lys His His Lys Ala Thr Gly His Met Thr Lys
145 150 155 160
gac cag gtc ttc gtc ccg aag acc cgc acc cag gtc ggc ctc ccg ccg 528
Asp Gln Val Phe Val Pro Lys Thr Arg Thr Gln Val Gly Leu Pro Pro
165 170 175
aag gag tcg gcc gcc gcc acc gtc cag gag gag gag gac atg tcg gtc 576
Lys Glu Ser Ala Ala Ala Thr Val Gln Glu Glu Glu Asp Met Ser Val
180 185 190
cac ctc gac gag gag gcc ccg atc gtc acc ctc ttc tgg atg gtc atc 624
His Leu Asp Glu Glu Ala Pro Ile Val Thr Leu Phe Trp Met Val Ile
195 200 205
cag ttc ctc ttc ggc tgg ccg gcc tac ctc atc atg aac gcc tcg ggc 672
Gln Phe Leu Phe Gly Trp Pro Ala Tyr Leu Ile Met Asn Ala Ser Gly
210 215 220
cag gac tac ggc cgc tgg acc tcg cac ttc cac acc tac tcg ccg atc 720
Gln Asp Tyr Gly Arg Trp Thr Ser His Phe His Thr Tyr Ser Pro Ile
225 230 235 240
ttc gag ccg cgc aac ttc ttc gac atc atc ctc tcg gac ctc ggc gtc 768
Phe Glu Pro Arg Asn Phe Phe Asp Ile Ile Leu Ser Asp Leu Gly Val
245 250 255
ctc gcc acc ctc ggc gcc ctc atc tac gcc tcg atg cag ctc tcg ctc 816
Leu Ala Thr Leu Gly Ala Leu Ile Tyr Ala Ser Met Gln Leu Ser Leu
260 265 270
ctc acc gtc acc aag tac tac atc atc ccg tac ctc ttc gtc aac ttc 864
Leu Thr Val Thr Lys Tyr Tyr Ile Ile Pro Tyr Leu Phe Val Asn Phe
275 280 285
tgg ctc gtc ctc atc acc ttc ctc cag cac acc gac ccg aag ctc ccg 912
Trp Leu Val Leu Ile Thr Phe Leu Gln His Thr Asp Pro Lys Leu Pro
290 295 300
cac tac cgc gag ggc gcc tgg aac ttc cag cgc ggc gcc ctc tgc acc 960
His Tyr Arg Glu Gly Ala Trp Asn Phe Gln Arg Gly Ala Leu Cys Thr
305 310 315 320
gtc gac cgc tcg ttc ggc aag ttc ctc gac cac atg ttc cac ggc atc 1008
Val Asp Arg Ser Phe Gly Lys Phe Leu Asp His Met Phe His Gly Ile
325 330 335
gtc cac acc cac gtc gcc cac cac ctc ttc tcg cag atg ccg ttc tac 1056
Val His Thr His Val Ala His His Leu Phe Ser Gln Met Pro Phe Tyr
340 345 350
cac gcc gag gag gcc acc tac cac ctc aag aag ctc ctc ggc gag tac 1104
His Ala Glu Glu Ala Thr Tyr His Leu Lys Lys Leu Leu Gly Glu Tyr
355 360 365
tac atc tac gac gcc tcg ccg atc gtc gtc gcc gtc tgg aag tcg ttc 1152
Tyr Ile Tyr Asp Ala Ser Pro Ile Val Val Ala Val Trp Lys Ser Phe
370 375 380
cgc gag tgc cgc ttc gtc gag gac cac ggc gac gtc gtc ttc ttc aag 1200
Arg Glu Cys Arg Phe Val Glu Asp His Gly Asp Val Val Phe Phe Lys
385 390 395 400
aag tag 1206
Lys
<210> 5
<211> 401
<212> PRT
<213> Mortierella alpine
<400> 5
Met Ala Pro Pro Asn Thr Ile Asp Ala Gly Leu Thr Gln Arg His Ile
1 5 10 15
Ser Thr Ser Ala Ala Pro Asn Ser Ala Lys Pro Thr Phe Glu Arg Asn
20 25 30
Tyr Gln Leu Pro Glu Phe Thr Ile Lys Glu Ile Arg Glu Cys Ile Pro
35 40 45
Ala His Cys Phe Glu Arg Ser Gly Leu Arg Gly Leu Cys His Val Ala
50 55 60
Ile Asp Leu Thr Trp Ala Ser Leu Leu Phe Leu Ala Ala Thr Gln Ile
65 70 75 80
Asp Lys Phe Glu Asn Pro Leu Ile Arg Tyr Leu Ala Trp Pro Val Tyr
85 90 95
Trp Ile Met Gln Gly Ile Val Cys Thr Gly Ile Trp Val Leu Ala His
100 105 110
Glu Cys Gly His Gln Ser Phe Ser Thr Ser Lys Thr Leu Asn Asn Thr
115 120 125
Val Gly Trp Ile Leu His Ser Met Leu Leu Val Pro Tyr His Ser Trp
130 135 140
Arg Ile Ser His Ser Lys His His Lys Ala Thr Gly His Met Thr Lys
145 150 155 160
Asp Gln Val Phe Val Pro Lys Thr Arg Thr Gln Val Gly Leu Pro Pro
165 170 175
Lys Glu Ser Ala Ala Ala Thr Val Gln Glu Glu Glu Asp Met Ser Val
180 185 190
His Leu Asp Glu Glu Ala Pro Ile Val Thr Leu Phe Trp Met Val Ile
195 200 205
Gln Phe Leu Phe Gly Trp Pro Ala Tyr Leu Ile Met Asn Ala Ser Gly
210 215 220
Gln Asp Tyr Gly Arg Trp Thr Ser His Phe His Thr Tyr Ser Pro Ile
225 230 235 240
Phe Glu Pro Arg Asn Phe Phe Asp Ile Ile Leu Ser Asp Leu Gly Val
245 250 255
Leu Ala Thr Leu Gly Ala Leu Ile Tyr Ala Ser Met Gln Leu Ser Leu
260 265 270
Leu Thr Val Thr Lys Tyr Tyr Ile Ile Pro Tyr Leu Phe Val Asn Phe
275 280 285
Trp Leu Val Leu Ile Thr Phe Leu Gln His Thr Asp Pro Lys Leu Pro
290 295 300
His Tyr Arg Glu Gly Ala Trp Asn Phe Gln Arg Gly Ala Leu Cys Thr
305 310 315 320
Val Asp Arg Ser Phe Gly Lys Phe Leu Asp His Met Phe His Gly Ile
325 330 335
Val His Thr His Val Ala His His Leu Phe Ser Gln Met Pro Phe Tyr
340 345 350
His Ala Glu Glu Ala Thr Tyr His Leu Lys Lys Leu Leu Gly Glu Tyr
355 360 365
Tyr Ile Tyr Asp Ala Ser Pro Ile Val Val Ala Val Trp Lys Ser Phe
370 375 380
Arg Glu Cys Arg Phe Val Glu Asp His Gly Asp Val Val Phe Phe Lys
385 390 395 400
Lys
<210> 6
<211> 2760
<212> DNA
<213> Rhodosporidium toruloides
<400> 6
caccgcttcc cggagaactt tgctgtactc tgcttctccc ttcacactct cacacccact 60
cacacaccct tccatccaca cacaagctat ccgcacacct ctcacacccg accccagctc 120
gccccatcct cttcgcaccc ggctcatcgc cacacacgca atgactgcct cgtcggcact 180
cgagacctcg ctcccgcact ctgtcgggcc cgagtctgcg actaccaccg caaagccgcc 240
ccgtgcgccg ctcaggatgc gtcaccccga ctacacccag accgacgtcc tcgagtcgtc 300
agactcggac gcagcgtcgg attccgaggg cgagacgacg gctgtcgacg acgggaccta 360
cgaggacgat aactacgtcc gcaaggtcct cagcaaggag aagccgctcc cgcccatcac 420
ctggaagaac atccaccgca acatccagtg gatctcgacc ctcgccctca ccatcgtgcc 480
cctcctctcg atctacggag cgttcacgac gcccctgaag tggcagacgg cggtctggag 540
tgtcgtctac tactacttca ccggtctcgg gtgagtcgcc gcaccctttc ctcgcctctc 600
ctccctcgct ctctacttcc ggcgcctccg ttcttcgttc atgatcgtct gtaagacgtg 660
tttgagcttg gaggggcgtc agatggcccg gcggacgagt cgcatgtcga gaaaccgtcg 720
ctgaccttct cttcgccttt tcctcctcct ccattgtccc cgatcgtccc gatccgtctg 780
ttctcgttct acagcatcac cgcaggctac cacaggctgt acgtgtcatt tactctgatc 840
tcgaggcgtt gagcgccgaa aaacggactc tgaactgacg aaaactcgct tttctcgcct 900
tcctcgcctg taggtgggcc cacaggtcct acactgcctc cctgcctctc cagtacttcc 960
tggcacttgg cggaagcggc gcagtggagg gttctgtgaa atggtgggcc cgaggacacc 1020
gcgcacacca ccgctacacc gacacggacc tcgacccgta ctcagcgcag aagggcttct 1080
ggtgggcaca cctcggctgg atgattgtca agccgcgccg tcgtcccggt gtcgccgatg 1140
tctccgacct caacaacaac ccagtcgtca agtggcagca ccgcttctac ctcccgctca 1200
tcctcggcat gggcttcatc ttccctacca tcgtcgctgg actcggctgg ggcgacttcc 1260
gcggcggatt tttcttcgcc ggcgctgctc gcctcctctt tgtccaccac gtgcgttctc 1320
gccttcctcg tctctcctcc ttcgttcgct gacgttttcg tattgcacag tcgacgttct 1380
gcgtcaactc gctcgcacac tggctcggcg agacgccgtt tgacgacaag cacacgccga 1440
aggaccactg gctcaccgcg ctcgcgacgg tcggcgaggg ctaccacaac ttcgtgagtc 1500
tcccgcccgc tctctgcgtg atctacggcg tcttcgtgta ctgctgccac agtcgcgacg 1560
tcgttcgagt cgtttgccat cgacttcgct ctcctcgacc tcccgctgac cttctccctt 1620
cccctccagc accacgagtt cccctccgac taccgcaacg cgctcagatg gtggcagtat 1680
gatccgacta agtgtttcat ttacgcgatg tcgaaactcg gattggcgtc gcagctcaag 1740
acgttccccg acaacgagat caagaagggt cagtacgcca tgacgctcaa ggctgtcgcg 1800
cgcgaggcgg agaacatcga gtggcccaag tcgtcgaacc acttgcctgt gctcacctgg 1860
gatgagtgta cgttcagctt tcatcgtcgt cgacggcgac ctcgaccttc ctgaacttcc 1920
agcgctgact ttcgtccccg ctcctctcgc tcgcagtcca ggaggcctgc aagactcgcc 1980
agctcctcgt tgtcgccggt ttcatccacg atgtcagcac cttcatcgac cagcaccctg 2040
gcggtgccgg cttgatcaag acccgtctcg gccgcgatgc gacgaccgcc ttctacggtg 2100
gctactacga gtgcgtcttc acattctcct gcatcatctt cgctcaagtc gctgacgctc 2160
tcgctcccgc agccactcga acggcgcagc caacttgctc gcccagtacc gtgtcggcgt 2220
catcgagggc ggctacgagg tcgagcacat gaagaagtac tctgaggtcg tcgagaacct 2280
caagaagcac ggcgccgacg gcgtggccgg caagagcgcc gacctcgcca agggtccgaa 2340
gcagatgtcg gtcatcaagg gcgaccctca gctcaagggc gcgccgctcg agacgctcgc 2400
caagccgcct accttcagcg agaccaacct tttgggcggt ctcagcctga cggtcaaggc 2460
gtaatggccc gtcaccgcca tcggttcaag ggagagtcgt cgaccgactc ttccgttcgt 2520
cgcttcaggt tcatccgtct tcgcatgctt agagaccctc ttcctcgttg ttttgtgcat 2580
cagttgcccg cagtcgtccc ttaacgagcc cactactacc ctttccatgc ccagtattct 2640
tgcccccgcc cccttcgttc tttgatacgt ccaaccacct cgctctttcg ctgtagctct 2700
acttcgttct ctcccttgcg ggttgcccgc ttactccgca tggaaacttg accagtgtgc 2760
<210> 7
<211> 1638
<212> DNA
<213> Rhodosporidium toruloides
<220>
<221> CDS
<222> (1)..(1638)
<400> 7
atg act gcc tcg tcg gca ctc gag acc tcg ctc ccg cac tct gtc ggg 48
Met Thr Ala Ser Ser Ala Leu Glu Thr Ser Leu Pro His Ser Val Gly
1 5 10 15
ccc gag tct gcg act acc acc gca aag ccg ccc cgt gcg ccg ctc agg 96
Pro Glu Ser Ala Thr Thr Thr Ala Lys Pro Pro Arg Ala Pro Leu Arg
20 25 30
atg cgt cac ccc gac tac acc cag acc gac gtc ctc gag tcg tca gac 144
Met Arg His Pro Asp Tyr Thr Gln Thr Asp Val Leu Glu Ser Ser Asp
35 40 45
tcg gac gca gcg tcg gat tcc gag ggc gag acg acg gct gtc gac gac 192
Ser Asp Ala Ala Ser Asp Ser Glu Gly Glu Thr Thr Ala Val Asp Asp
50 55 60
ggg acc tac gag gac gac aac tac gtc cgc aag gtc ctc agc aag gag 240
Gly Thr Tyr Glu Asp Asp Asn Tyr Val Arg Lys Val Leu Ser Lys Glu
65 70 75 80
aag ccg ctc ccg ccc atc acc tgg aag aac atc cac cgc aac atc cag 288
Lys Pro Leu Pro Pro Ile Thr Trp Lys Asn Ile His Arg Asn Ile Gln
85 90 95
tgg atc tcg acc ctc gcc ctc acc atc gtg ccc ctc ctc tcg atc tac 336
Trp Ile Ser Thr Leu Ala Leu Thr Ile Val Pro Leu Leu Ser Ile Tyr
100 105 110
gga gcg ttc acg acg ccc ctg aag tgg cag acg gcg gtc tgg agt gtc 384
Gly Ala Phe Thr Thr Pro Leu Lys Trp Gln Thr Ala Val Trp Ser Val
115 120 125
gtc tac tac tac ttc acc ggt ctc ggc atc acc gca ggc tac cac agg 432
Val Tyr Tyr Tyr Phe Thr Gly Leu Gly Ile Thr Ala Gly Tyr His Arg
130 135 140
ctg tgg gcc cac agg tcc tac act gcc tcc ctg cct ctc cag tac ttc 480
Leu Trp Ala His Arg Ser Tyr Thr Ala Ser Leu Pro Leu Gln Tyr Phe
145 150 155 160
ctg gca ctt ggc gga agc ggc gca gtc gag ggt tct gtg aaa tgg tgg 528
Leu Ala Leu Gly Gly Ser Gly Ala Val Glu Gly Ser Val Lys Trp Trp
165 170 175
gcc cga gga cac cgc gca cac cac cgc tac acc gac acg gac ctc gac 576
Ala Arg Gly His Arg Ala His His Arg Tyr Thr Asp Thr Asp Leu Asp
180 185 190
ccg tac tca gcg cag aag ggc ttc tgg tgg gca cac ctc ggc tgg atg 624
Pro Tyr Ser Ala Gln Lys Gly Phe Trp Trp Ala His Leu Gly Trp Met
195 200 205
att gtc aag ccg cgc cgt cgt ccc ggt gtc gcc gat gtc tcc gac ctc 672
Ile Val Lys Pro Arg Arg Arg Pro Gly Val Ala Asp Val Ser Asp Leu
210 215 220
aac aac aac cca gtc gtc aag tgg cag cac cgc ttc tac ctc ccg ctc 720
Asn Asn Asn Pro Val Val Lys Trp Gln His Arg Phe Tyr Leu Pro Leu
225 230 235 240
atc ctc ggc atg ggc ttc atc ttc cct acc atc gtc gct gga ctc ggc 768
Ile Leu Gly Met Gly Phe Ile Phe Pro Thr Ile Val Ala Gly Leu Gly
245 250 255
tgg ggc gac ttc cgc ggc gga ttt ttc ttc gcc ggc gct gct cgc ctc 816
Trp Gly Asp Phe Arg Gly Gly Phe Phe Phe Ala Gly Ala Ala Arg Leu
260 265 270
ctc ttt gtc cac cac tcg acg ttc tgc gtc aac tcg ctc gca cac tgg 864
Leu Phe Val His His Ser Thr Phe Cys Val Asn Ser Leu Ala His Trp
275 280 285
ctc ggc gag acg ccg ttt gac gac aag cac acg ccg aag gac cac tgg 912
Leu Gly Glu Thr Pro Phe Asp Asp Lys His Thr Pro Lys Asp His Trp
290 295 300
ctc acc gcg ctc gcg acg gtc ggc gag ggc tac cac aac ttc cac cac 960
Leu Thr Ala Leu Ala Thr Val Gly Glu Gly Tyr His Asn Phe His His
305 310 315 320
gag ttc ccc tcc gac tac cgc aac gcg ctc aga tgg tgg cag tat gat 1008
Glu Phe Pro Ser Asp Tyr Arg Asn Ala Leu Arg Trp Trp Gln Tyr Asp
325 330 335
ccg act aag tgt ttc att tac gcg atg tcg aaa ctc gga ttg gcg tcg 1056
Pro Thr Lys Cys Phe Ile Tyr Ala Met Ser Lys Leu Gly Leu Ala Ser
340 345 350
cag ctc aag acg ttc ccc gac aac gag atc aag aag ggt cag tac gcc 1104
Gln Leu Lys Thr Phe Pro Asp Asn Glu Ile Lys Lys Gly Gln Tyr Ala
355 360 365
atg acg ctc aag gct gtc gcg cgc gag gcg gag aac atc gag tgg ccc 1152
Met Thr Leu Lys Ala Val Ala Arg Glu Ala Glu Asn Ile Glu Trp Pro
370 375 380
aag tcg tcg aac cac ttg cct gtg ctc acc tgg gat gag ttc cag gag 1200
Lys Ser Ser Asn His Leu Pro Val Leu Thr Trp Asp Glu Phe Gln Glu
385 390 395 400
gcc tgc aag act cgc cag ctc ctc gtt gtc gcc ggt ttc atc cac gat 1248
Ala Cys Lys Thr Arg Gln Leu Leu Val Val Ala Gly Phe Ile His Asp
405 410 415
gtc agc acc ttc atc gac cag cac cct ggc ggt gcc ggc ttg atc aag 1296
Val Ser Thr Phe Ile Asp Gln His Pro Gly Gly Ala Gly Leu Ile Lys
420 425 430
acc cgt ctc ggc cgc gat gcg acg acc gcc ttc tac ggt ggc tac tac 1344
Thr Arg Leu Gly Arg Asp Ala Thr Thr Ala Phe Tyr Gly Gly Tyr Tyr
435 440 445
gac cac tcg aac ggc gca gcc aac ttg ctc gcc cag tac cgt gtc ggc 1392
Asp His Ser Asn Gly Ala Ala Asn Leu Leu Ala Gln Tyr Arg Val Gly
450 455 460
gtc atc gag ggc ggc tac gag gtc gag cac atg aag aag tac tct gag 1440
Val Ile Glu Gly Gly Tyr Glu Val Glu His Met Lys Lys Tyr Ser Glu
465 470 475 480
gtc gtc gag aac ctc aag aag cac ggc gcc gac ggc gtg gcc ggc aag 1488
Val Val Glu Asn Leu Lys Lys His Gly Ala Asp Gly Val Ala Gly Lys
485 490 495
agc gcc gac ctc gcc aag ggt ccg aag cag atg tcg gtc atc aag ggc 1536
Ser Ala Asp Leu Ala Lys Gly Pro Lys Gln Met Ser Val Ile Lys Gly
500 505 510
gac cct cag ctc aag ggc gcg ccg ctc gag acg ctc gcc aag ccg cct 1584
Asp Pro Gln Leu Lys Gly Ala Pro Leu Glu Thr Leu Ala Lys Pro Pro
515 520 525
acc ttc agc gag acc aac ctt ttg ggc ggt ctc agc ctg acg gtc aag 1632
Thr Phe Ser Glu Thr Asn Leu Leu Gly Gly Leu Ser Leu Thr Val Lys
530 535 540
gcg taa 1638
Ala
545
<210> 8
<211> 545
<212> PRT
<213> Rhodosporidium toruloides
<400> 8
Met Thr Ala Ser Ser Ala Leu Glu Thr Ser Leu Pro His Ser Val Gly
1 5 10 15
Pro Glu Ser Ala Thr Thr Thr Ala Lys Pro Pro Arg Ala Pro Leu Arg
20 25 30
Met Arg His Pro Asp Tyr Thr Gln Thr Asp Val Leu Glu Ser Ser Asp
35 40 45
Ser Asp Ala Ala Ser Asp Ser Glu Gly Glu Thr Thr Ala Val Asp Asp
50 55 60
Gly Thr Tyr Glu Asp Asp Asn Tyr Val Arg Lys Val Leu Ser Lys Glu
65 70 75 80
Lys Pro Leu Pro Pro Ile Thr Trp Lys Asn Ile His Arg Asn Ile Gln
85 90 95
Trp Ile Ser Thr Leu Ala Leu Thr Ile Val Pro Leu Leu Ser Ile Tyr
100 105 110
Gly Ala Phe Thr Thr Pro Leu Lys Trp Gln Thr Ala Val Trp Ser Val
115 120 125
Val Tyr Tyr Tyr Phe Thr Gly Leu Gly Ile Thr Ala Gly Tyr His Arg
130 135 140
Leu Trp Ala His Arg Ser Tyr Thr Ala Ser Leu Pro Leu Gln Tyr Phe
145 150 155 160
Leu Ala Leu Gly Gly Ser Gly Ala Val Glu Gly Ser Val Lys Trp Trp
165 170 175
Ala Arg Gly His Arg Ala His His Arg Tyr Thr Asp Thr Asp Leu Asp
180 185 190
Pro Tyr Ser Ala Gln Lys Gly Phe Trp Trp Ala His Leu Gly Trp Met
195 200 205
Ile Val Lys Pro Arg Arg Arg Pro Gly Val Ala Asp Val Ser Asp Leu
210 215 220
Asn Asn Asn Pro Val Val Lys Trp Gln His Arg Phe Tyr Leu Pro Leu
225 230 235 240
Ile Leu Gly Met Gly Phe Ile Phe Pro Thr Ile Val Ala Gly Leu Gly
245 250 255
Trp Gly Asp Phe Arg Gly Gly Phe Phe Phe Ala Gly Ala Ala Arg Leu
260 265 270
Leu Phe Val His His Ser Thr Phe Cys Val Asn Ser Leu Ala His Trp
275 280 285
Leu Gly Glu Thr Pro Phe Asp Asp Lys His Thr Pro Lys Asp His Trp
290 295 300
Leu Thr Ala Leu Ala Thr Val Gly Glu Gly Tyr His Asn Phe His His
305 310 315 320
Glu Phe Pro Ser Asp Tyr Arg Asn Ala Leu Arg Trp Trp Gln Tyr Asp
325 330 335
Pro Thr Lys Cys Phe Ile Tyr Ala Met Ser Lys Leu Gly Leu Ala Ser
340 345 350
Gln Leu Lys Thr Phe Pro Asp Asn Glu Ile Lys Lys Gly Gln Tyr Ala
355 360 365
Met Thr Leu Lys Ala Val Ala Arg Glu Ala Glu Asn Ile Glu Trp Pro
370 375 380
Lys Ser Ser Asn His Leu Pro Val Leu Thr Trp Asp Glu Phe Gln Glu
385 390 395 400
Ala Cys Lys Thr Arg Gln Leu Leu Val Val Ala Gly Phe Ile His Asp
405 410 415
Val Ser Thr Phe Ile Asp Gln His Pro Gly Gly Ala Gly Leu Ile Lys
420 425 430
Thr Arg Leu Gly Arg Asp Ala Thr Thr Ala Phe Tyr Gly Gly Tyr Tyr
435 440 445
Asp His Ser Asn Gly Ala Ala Asn Leu Leu Ala Gln Tyr Arg Val Gly
450 455 460
Val Ile Glu Gly Gly Tyr Glu Val Glu His Met Lys Lys Tyr Ser Glu
465 470 475 480
Val Val Glu Asn Leu Lys Lys His Gly Ala Asp Gly Val Ala Gly Lys
485 490 495
Ser Ala Asp Leu Ala Lys Gly Pro Lys Gln Met Ser Val Ile Lys Gly
500 505 510
Asp Pro Gln Leu Lys Gly Ala Pro Leu Glu Thr Leu Ala Lys Pro Pro
515 520 525
Thr Phe Ser Glu Thr Asn Leu Leu Gly Gly Leu Ser Leu Thr Val Lys
530 535 540
Ala
545
<210> 9
<211> 1164
<212> DNA
<213> Linum usitatissimum
<220>
<221> CDS
<222> (1)..(1164)
<400> 9
atg aag cag cag cag tac aag gac acc ccg atc ctc aac ggc gtc aac 48
Met Lys Gln Gln Gln Tyr Lys Asp Thr Pro Ile Leu Asn Gly Val Asn
1 5 10 15
ggc ttc cac gcc aag gag gag gag gag gag gag gac ttc gac ctc tcg 96
Gly Phe His Ala Lys Glu Glu Glu Glu Glu Glu Asp Phe Asp Leu Ser
20 25 30
aac ccg ccg ccg ttc aac atc ggc cag atc cgc gcc gcc atc ccg aag 144
Asn Pro Pro Pro Phe Asn Ile Gly Gln Ile Arg Ala Ala Ile Pro Lys
35 40 45
cac tgc tgg gtc aag aac ccg tgg cgc tcg ctc acc tac gtc ttc cgc 192
His Cys Trp Val Lys Asn Pro Trp Arg Ser Leu Thr Tyr Val Phe Arg
50 55 60
gac gtc gtc gtc gtc ttc gcc ctc gcc gcc gcc gcc ttc tac ttc aac 240
Asp Val Val Val Val Phe Ala Leu Ala Ala Ala Ala Phe Tyr Phe Asn
65 70 75 80
tcg tgg ctc ttc tgg ccg ctc tac tgg ttc gcc cag ggc acc atg ttc 288
Ser Trp Leu Phe Trp Pro Leu Tyr Trp Phe Ala Gln Gly Thr Met Phe
85 90 95
tgg gcc atc ttc gtc ctc ggc cac gac tgc ggc cac ggc tcg ttc tcg 336
Trp Ala Ile Phe Val Leu Gly His Asp Cys Gly His Gly Ser Phe Ser
100 105 110
aac aac tcg tcg ctc aac aac gtc gtc ggc cac ctc ctc cac tcg tcg 384
Asn Asn Ser Ser Leu Asn Asn Val Val Gly His Leu Leu His Ser Ser
115 120 125
atc ctc gtc ccg tac cac ggc tgg cgc atc tcg cac cgc acc cac cac 432
Ile Leu Val Pro Tyr His Gly Trp Arg Ile Ser His Arg Thr His His
130 135 140
cag aac cac ggc aac gtc gag aag gac gag tcg tgg gtc ccg ctc ccg 480
Gln Asn His Gly Asn Val Glu Lys Asp Glu Ser Trp Val Pro Leu Pro
145 150 155 160
gag aag atc tac aag gag atg gac ctc tcg acc cgc atc ctc cgc tac 528
Glu Lys Ile Tyr Lys Glu Met Asp Leu Ser Thr Arg Ile Leu Arg Tyr
165 170 175
tcg gtc ccg ctc ccg atg ttc gcc ctc ccg ttc tac ctc tgg tgg cgc 576
Ser Val Pro Leu Pro Met Phe Ala Leu Pro Phe Tyr Leu Trp Trp Arg
180 185 190
tcg ccg ggc aag gag ggc tcg cac ttc aac ccg aac tcg gac ttc ttc 624
Ser Pro Gly Lys Glu Gly Ser His Phe Asn Pro Asn Ser Asp Phe Phe
195 200 205
gcc ccg cac gag cgc aag gcc gtc ctc acc tcg aac ttc tgc ttc tcg 672
Ala Pro His Glu Arg Lys Ala Val Leu Thr Ser Asn Phe Cys Phe Ser
210 215 220
atc atg gcc ctc ctc ctc ctc tac tcg tgc ttc gtc ttc ggc ccg gtc 720
Ile Met Ala Leu Leu Leu Leu Tyr Ser Cys Phe Val Phe Gly Pro Val
225 230 235 240
cag gtc ctc aag ttc tac ggc atc ccg tac ctc gtc ttc gtc atg tgg 768
Gln Val Leu Lys Phe Tyr Gly Ile Pro Tyr Leu Val Phe Val Met Trp
245 250 255
ctc gac ttc gtc acc tac atg cac cac cac ggc cac gag gag aag ctc 816
Leu Asp Phe Val Thr Tyr Met His His His Gly His Glu Glu Lys Leu
260 265 270
ccg tgg tac cgc ggc aag gag tgg tcg tac ctc cgc ggc ggc ctc acc 864
Pro Trp Tyr Arg Gly Lys Glu Trp Ser Tyr Leu Arg Gly Gly Leu Thr
275 280 285
acc gtc gac cgc gac tac ggc tgg atc aac aac atc cac cac gac atc 912
Thr Val Asp Arg Asp Tyr Gly Trp Ile Asn Asn Ile His His Asp Ile
290 295 300
ggc acc cac gtc atc cac cac ctc ttc ccg cag atc ccg cac tac cac 960
Gly Thr His Val Ile His His Leu Phe Pro Gln Ile Pro His Tyr His
305 310 315 320
ctc atc gag gcc acc aag gcc gcc aag ccg gtc ctc ggc aag tac tac 1008
Leu Ile Glu Ala Thr Lys Ala Ala Lys Pro Val Leu Gly Lys Tyr Tyr
325 330 335
cgc gag ccg aag aag tcg ggc ccg ttc ccg ttc cac ctc ttc tcg aac 1056
Arg Glu Pro Lys Lys Ser Gly Pro Phe Pro Phe His Leu Phe Ser Asn
340 345 350
ctc gtc cgc tcg atg tcg gag gac cac tac gtc tcg gac atc ggc gac 1104
Leu Val Arg Ser Met Ser Glu Asp His Tyr Val Ser Asp Ile Gly Asp
355 360 365
atc gtc ttc tac cag acc gac ccg gac atc tac aag gtc gac aag tcg 1152
Ile Val Phe Tyr Gln Thr Asp Pro Asp Ile Tyr Lys Val Asp Lys Ser
370 375 380
aag ctc aac tag 1164
Lys Leu Asn
385
<210> 10
<211> 387
<212> PRT
<213> Linum usitatissimum
<400> 10
Met Lys Gln Gln Gln Tyr Lys Asp Thr Pro Ile Leu Asn Gly Val Asn
1 5 10 15
Gly Phe His Ala Lys Glu Glu Glu Glu Glu Glu Asp Phe Asp Leu Ser
20 25 30
Asn Pro Pro Pro Phe Asn Ile Gly Gln Ile Arg Ala Ala Ile Pro Lys
35 40 45
His Cys Trp Val Lys Asn Pro Trp Arg Ser Leu Thr Tyr Val Phe Arg
50 55 60
Asp Val Val Val Val Phe Ala Leu Ala Ala Ala Ala Phe Tyr Phe Asn
65 70 75 80
Ser Trp Leu Phe Trp Pro Leu Tyr Trp Phe Ala Gln Gly Thr Met Phe
85 90 95
Trp Ala Ile Phe Val Leu Gly His Asp Cys Gly His Gly Ser Phe Ser
100 105 110
Asn Asn Ser Ser Leu Asn Asn Val Val Gly His Leu Leu His Ser Ser
115 120 125
Ile Leu Val Pro Tyr His Gly Trp Arg Ile Ser His Arg Thr His His
130 135 140
Gln Asn His Gly Asn Val Glu Lys Asp Glu Ser Trp Val Pro Leu Pro
145 150 155 160
Glu Lys Ile Tyr Lys Glu Met Asp Leu Ser Thr Arg Ile Leu Arg Tyr
165 170 175
Ser Val Pro Leu Pro Met Phe Ala Leu Pro Phe Tyr Leu Trp Trp Arg
180 185 190
Ser Pro Gly Lys Glu Gly Ser His Phe Asn Pro Asn Ser Asp Phe Phe
195 200 205
Ala Pro His Glu Arg Lys Ala Val Leu Thr Ser Asn Phe Cys Phe Ser
210 215 220
Ile Met Ala Leu Leu Leu Leu Tyr Ser Cys Phe Val Phe Gly Pro Val
225 230 235 240
Gln Val Leu Lys Phe Tyr Gly Ile Pro Tyr Leu Val Phe Val Met Trp
245 250 255
Leu Asp Phe Val Thr Tyr Met His His His Gly His Glu Glu Lys Leu
260 265 270
Pro Trp Tyr Arg Gly Lys Glu Trp Ser Tyr Leu Arg Gly Gly Leu Thr
275 280 285
Thr Val Asp Arg Asp Tyr Gly Trp Ile Asn Asn Ile His His Asp Ile
290 295 300
Gly Thr His Val Ile His His Leu Phe Pro Gln Ile Pro His Tyr His
305 310 315 320
Leu Ile Glu Ala Thr Lys Ala Ala Lys Pro Val Leu Gly Lys Tyr Tyr
325 330 335
Arg Glu Pro Lys Lys Ser Gly Pro Phe Pro Phe His Leu Phe Ser Asn
340 345 350
Leu Val Arg Ser Met Ser Glu Asp His Tyr Val Ser Asp Ile Gly Asp
355 360 365
Ile Val Phe Tyr Gln Thr Asp Pro Asp Ile Tyr Lys Val Asp Lys Ser
370 375 380
Lys Leu Asn
385
<210> 11
<211> 1164
<212> DNA
<213> Vernicia fordii
<220>
<221> CDS
<222> (1)..(1164)
<400> 11
atg aag cag cag cag tac aag gac acc ccg atc ctc aac ggc gtc aac 48
Met Lys Gln Gln Gln Tyr Lys Asp Thr Pro Ile Leu Asn Gly Val Asn
1 5 10 15
ggc ttc cac gcc aag gag gag gag gag gag gag gac ttc gac ctc tcg 96
Gly Phe His Ala Lys Glu Glu Glu Glu Glu Glu Asp Phe Asp Leu Ser
20 25 30
aac ccg ccg ccg ttc aac atc ggc cag atc cgc gcc gcc atc ccg aag 144
Asn Pro Pro Pro Phe Asn Ile Gly Gln Ile Arg Ala Ala Ile Pro Lys
35 40 45
cac tgc tgg gtc aag aac ccg tgg cgc tcg ctc acc tac gtc ttc cgc 192
His Cys Trp Val Lys Asn Pro Trp Arg Ser Leu Thr Tyr Val Phe Arg
50 55 60
gac gtc gtc gtc gtc ttc gcc ctc gcc gcc gcc gcc ttc tac ttc aac 240
Asp Val Val Val Val Phe Ala Leu Ala Ala Ala Ala Phe Tyr Phe Asn
65 70 75 80
tcg tgg ctc ttc tgg ccg ctc tac tgg ttc gcc cag ggc acc atg ttc 288
Ser Trp Leu Phe Trp Pro Leu Tyr Trp Phe Ala Gln Gly Thr Met Phe
85 90 95
tgg gcc atc ttc gtc ctc ggc cac gac tgc ggc cac ggc tcg ttc tcg 336
Trp Ala Ile Phe Val Leu Gly His Asp Cys Gly His Gly Ser Phe Ser
100 105 110
aac aac tcg tcg ctc aac aac gtc gtc ggc cac ctc ctc cac tcg tcg 384
Asn Asn Ser Ser Leu Asn Asn Val Val Gly His Leu Leu His Ser Ser
115 120 125
atc ctc gtc ccg tac cac ggc tgg cgc atc tcg cac cgc acc cac cac 432
Ile Leu Val Pro Tyr His Gly Trp Arg Ile Ser His Arg Thr His His
130 135 140
cag aac cac ggc aac gtc gag aag gac gag tcg tgg gtc ccg ctc ccg 480
Gln Asn His Gly Asn Val Glu Lys Asp Glu Ser Trp Val Pro Leu Pro
145 150 155 160
gag aag atc tac aag gag atg gac ctc tcg acc cgc atc ctc cgc tac 528
Glu Lys Ile Tyr Lys Glu Met Asp Leu Ser Thr Arg Ile Leu Arg Tyr
165 170 175
tcg gtc ccg ctc ccg atg ttc gcc ctc ccg ttc tac ctc tgg tgg cgc 576
Ser Val Pro Leu Pro Met Phe Ala Leu Pro Phe Tyr Leu Trp Trp Arg
180 185 190
tcg ccg ggc aag gag ggc tcg cac ttc aac ccg aac tcg gac ttc ttc 624
Ser Pro Gly Lys Glu Gly Ser His Phe Asn Pro Asn Ser Asp Phe Phe
195 200 205
gcc ccg cac gag cgc aag gcc gtc ctc acc tcg aac ttc tgc ttc tcg 672
Ala Pro His Glu Arg Lys Ala Val Leu Thr Ser Asn Phe Cys Phe Ser
210 215 220
atc atg gcc ctc ctc ctc ctc tac tcg tgc ttc gtc ttc ggc ccg gtc 720
Ile Met Ala Leu Leu Leu Leu Tyr Ser Cys Phe Val Phe Gly Pro Val
225 230 235 240
cag gtc ctc aag ttc tac ggc atc ccg tac ctc gtc ttc gtc atg tgg 768
Gln Val Leu Lys Phe Tyr Gly Ile Pro Tyr Leu Val Phe Val Met Trp
245 250 255
ctc gac ttc gtc acc tac atg cac cac cac ggc cac gag gag aag ctc 816
Leu Asp Phe Val Thr Tyr Met His His His Gly His Glu Glu Lys Leu
260 265 270
ccg tgg tac cgc ggc aag gag tgg tcg tac ctc cgc ggc ggc ctc acc 864
Pro Trp Tyr Arg Gly Lys Glu Trp Ser Tyr Leu Arg Gly Gly Leu Thr
275 280 285
acc gtc gac cgc gac tac ggc tgg atc aac aac atc cac cac gac atc 912
Thr Val Asp Arg Asp Tyr Gly Trp Ile Asn Asn Ile His His Asp Ile
290 295 300
ggc acc cac gtc atc cac cac ctc ttc ccg cag atc ccg cac tac cac 960
Gly Thr His Val Ile His His Leu Phe Pro Gln Ile Pro His Tyr His
305 310 315 320
ctc atc gag gcc acc aag gcc gcc aag ccg gtc ctc ggc aag tac tac 1008
Leu Ile Glu Ala Thr Lys Ala Ala Lys Pro Val Leu Gly Lys Tyr Tyr
325 330 335
cgc gag ccg aag aag tcg ggc ccg ttc ccg ttc cac ctc ttc tcg aac 1056
Arg Glu Pro Lys Lys Ser Gly Pro Phe Pro Phe His Leu Phe Ser Asn
340 345 350
ctc gtc cgc tcg atg tcg gag gac cac tac gtc tcg gac atc ggc gac 1104
Leu Val Arg Ser Met Ser Glu Asp His Tyr Val Ser Asp Ile Gly Asp
355 360 365
atc gtc ttc tac cag acc gac ccg gac atc tac aag gtc gac aag tcg 1152
Ile Val Phe Tyr Gln Thr Asp Pro Asp Ile Tyr Lys Val Asp Lys Ser
370 375 380
aag ctc aac tag 1164
Lys Leu Asn
385
<210> 12
<211> 387
<212> PRT
<213> Vernicia fordii
<400> 12
Met Lys Gln Gln Gln Tyr Lys Asp Thr Pro Ile Leu Asn Gly Val Asn
1 5 10 15
Gly Phe His Ala Lys Glu Glu Glu Glu Glu Glu Asp Phe Asp Leu Ser
20 25 30
Asn Pro Pro Pro Phe Asn Ile Gly Gln Ile Arg Ala Ala Ile Pro Lys
35 40 45
His Cys Trp Val Lys Asn Pro Trp Arg Ser Leu Thr Tyr Val Phe Arg
50 55 60
Asp Val Val Val Val Phe Ala Leu Ala Ala Ala Ala Phe Tyr Phe Asn
65 70 75 80
Ser Trp Leu Phe Trp Pro Leu Tyr Trp Phe Ala Gln Gly Thr Met Phe
85 90 95
Trp Ala Ile Phe Val Leu Gly His Asp Cys Gly His Gly Ser Phe Ser
100 105 110
Asn Asn Ser Ser Leu Asn Asn Val Val Gly His Leu Leu His Ser Ser
115 120 125
Ile Leu Val Pro Tyr His Gly Trp Arg Ile Ser His Arg Thr His His
130 135 140
Gln Asn His Gly Asn Val Glu Lys Asp Glu Ser Trp Val Pro Leu Pro
145 150 155 160
Glu Lys Ile Tyr Lys Glu Met Asp Leu Ser Thr Arg Ile Leu Arg Tyr
165 170 175
Ser Val Pro Leu Pro Met Phe Ala Leu Pro Phe Tyr Leu Trp Trp Arg
180 185 190
Ser Pro Gly Lys Glu Gly Ser His Phe Asn Pro Asn Ser Asp Phe Phe
195 200 205
Ala Pro His Glu Arg Lys Ala Val Leu Thr Ser Asn Phe Cys Phe Ser
210 215 220
Ile Met Ala Leu Leu Leu Leu Tyr Ser Cys Phe Val Phe Gly Pro Val
225 230 235 240
Gln Val Leu Lys Phe Tyr Gly Ile Pro Tyr Leu Val Phe Val Met Trp
245 250 255
Leu Asp Phe Val Thr Tyr Met His His His Gly His Glu Glu Lys Leu
260 265 270
Pro Trp Tyr Arg Gly Lys Glu Trp Ser Tyr Leu Arg Gly Gly Leu Thr
275 280 285
Thr Val Asp Arg Asp Tyr Gly Trp Ile Asn Asn Ile His His Asp Ile
290 295 300
Gly Thr His Val Ile His His Leu Phe Pro Gln Ile Pro His Tyr His
305 310 315 320
Leu Ile Glu Ala Thr Lys Ala Ala Lys Pro Val Leu Gly Lys Tyr Tyr
325 330 335
Arg Glu Pro Lys Lys Ser Gly Pro Phe Pro Phe His Leu Phe Ser Asn
340 345 350
Leu Val Arg Ser Met Ser Glu Asp His Tyr Val Ser Asp Ile Gly Asp
355 360 365
Ile Val Phe Tyr Gln Thr Asp Pro Asp Ile Tyr Lys Val Asp Lys Ser
370 375 380
Lys Leu Asn
385
<210> 13
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> Oligonucleotide
<400> 13
tttccgcggt cgaatttccc cgatcgttca 30
<210> 14
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 14
gagtcgctca cctactgcat c 21
<210> 15
<211> 31
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 15
tttgtttaaa catgctaatt cgggggatct g 31
<210> 16
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 16
cacgtcgact gaaacgcag 19
<210> 17
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 17
gacgaggtca tccgcgag 18
<210> 18
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 18
gtgcgggtcg tgatggac 18
<210> 19
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 19
ctggaaggcg tacgaggac 19
<210> 20
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 20
gtcaagccgc ccaggctgtc 20
<210> 21
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 21
ggatccgcca agtcgcgcag 20
<210> 22
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 22
gagtcgctca cctactgcat c 21
<210> 23
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 23
gagaactcgc cgatgtcgag 20
<210> 24
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 24
agcgactggt agagctggtc 20
<210> 25
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 25
ctcttgacga cacggcttac 20
<210> 26
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 26
caggccgcag agggtgaac 19
<210> 27
<211> 26
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 27
gatgagtttg gacaaaccac aactag 26
<210> 28
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 28
ggttcagggg gagatgtggg ag 22
<210> 29
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 29
gtaccggcgc gcccacctg 19
<210> 30
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 30
gaatcctgtt gccggtcttg cgatg 25
<210> 31
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 31
ttatgattag agtcccgcaa ttataca 27
<210> 32
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 32
ctagcttagc ttgagcttgg atc 23
<210> 33
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 33
gtgctgacgc gggcatagcc cag 23
<210> 34
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 34
atgcgactaa aacacgcgac aaga 24
<210> 35
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 35
agcagcggag gggttggatc 20
<210> 36
<211> 37
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<220>
<221> misc_feature
<222> (27)..(27)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (31)..(33)
<223> n is a, c, g, or t
<400> 36
acgatggact ccagagcggc cgcgcangca nnnggaa 37
<210> 37
<211> 40
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<220>
<221> misc_feature
<222> (30)..(30)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (34)..(36)
<223> n is a, c, g, or t
<400> 37
acgatggact ccagagcggc cgcgctgatn gctnnncggt 40
<210> 38
<211> 16
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 38
acgatggact ccagag 16
<210> 39
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 39
catacaccgg gcaaagcag 19
<210> 40
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 40
tttccatggg ccagcaggcg acgc 24
<210> 41
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 41
tttgatatca ggcgataata ttgagctcc 29
<210> 42
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 42
tttccatggg cgggagtgaa gggttgcc 28
<210> 43
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 43
tttgagatcc tactgcgcct gctgctcc 28
<210> 44
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 44
tttactagtc tcttcagacg gcttgttctc 30
<210> 45
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 45
cacccgtcct ctccgcttc 19
<210> 46
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 46
cctcgctctt tcgctggttc 20
<210> 47
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 47
ccgccaataa cctcacctca g 21
<210> 48
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 48
ggcgatggga gcgtagaata c 21
<210> 49
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 49
caggtttcat cgcaactaca ttga 24
<210> 50
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 50
aacagagcga gttgaagagt agcc 24
<210> 51
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 51
ccgccaataa cctcacctca g 21
<210> 52
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> oligonucleotide
<400> 52
gaagaagacg ccgagtagga tg 22
<210> 53
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> peptide motif
<220>
<221> misc_feature
<222> (2)..(2)
<223> Xaa can be any naturally occurring amino acid
<220>
<221> misc_feature
<222> (4)..(5)
<223> Xaa can be any naturally occurring amino acid
<400> 53
Gly Xaa Gly Xaa Xaa Gly
1 5
<210> 54
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> peptide motif
<400> 54
Gly Ser Gly Thr Val Gly
1 5
<210> 55
<211> 597
<212> DNA
<213> Ustilago maydis
<400> 55
gattagatct tgctgatagg caggtttgct tggagaatgg ggggaaaaga ctgaccgaag 60
aaacagcgag atctagaagt gataagcgga aagaatctga cttgctgtga tcagcagcca 120
attttttttt cgtttttttt ttttcactcc acatcgtcgt gcgtgcacgg tctgcatgtg 180
taaattgtat tcatcgaaag ccacagttga atacatcagc ccgatgtgga tttcgaaaac 240
caattaatct tggaattcac gcgctcagat cagtccatag agtcgacttc ggctgtttcc 300
aagagcttct tctctgcgag gtggttgccc gtgtttctcg ctgggaaaaa aggatcgatt 360
attattcgct tctacctcgc tcgcaccctt ggcctgctga aggaaacagc gccgagactc 420
ggtcacggtt gctgggctcc gtgttgatgc tgggacggcg caaagtgggg cccgcgcact 480
cttcgagcca aggacctcac tcttcaagaa caagcgctgt cgccatcgtc ttcttctttc 540
tgctccacca tcgaatcttt ctttctcgtt tcgaaaccaa aacactcttc caccatg 597
<210> 56
<211> 1434
<212> DNA
<213> Rhodosporidium toruloides
<400> 56
ctgcagaact acgccctctc acacccaact tccgactcga ccggcggtac gagcacgacc 60
tacttctact gcctgccatc gacatccggg cgggtcgctg cctaccctgt gcgttctgcg 120
ccctccctcg tctcgggagg cagtgtctga cagaagcttt gcgcgcagta ccccgtcaag 180
atgcaactct acgcaacgtt cggcacagaa gtcgccaagc tccgcgcatc gccgcctcaa 240
gctctcgcgc tgcccgacgg tgtcgtctat tacgaggcgg agaagctcga gttgccggct 300
ttgccagcgg cggtcaaggt tgaggtggag acggagaagg cgggagtagc gggggaggac 360
aatgaggcga agggtgagat ggtgctggtg gagactctta cggtggagca ggaggagatt 420
gaattgggct cgggagtcgt gcagattgag gagtcgttgc tcgtcaagct ggaggtcagc 480
ggctgatcct tccgttcgtt gcaaggatcg tctgcatgtt tcgcttctct caatgacaca 540
acctggagag cgctcccgtc agcgagaatc gaggacattc cgcagctcgt gagcaagcgg 600
aggtgcgagg ctccctcgaa agctgcgcct cttcagacgg cttgttctct cctgctctgg 660
tgggctggcc tgacatgtaa tgtgctccgc cgcaagtccg tcgtcggtct caattcgacg 720
ttgaaagggc atagcgcaag gaagaaccct ctgcggacat gcagaattac tggctcgcct 780
gctccttcgt ctactggaat aagtcctgtc tcgttaaagc cccaacgtcg tttttcgacg 840
tttgtaaggc gcaagaggtg ctatgggcta cgcaggaagc tgagaggaca tagaagtcgg 900
gggaggaacg gcgcagagcg gcagttgcgg aagcatgagg aaagcgagac ggtccagcat 960
ctgcagcgcc aatccgcaat ctcctggttg agcctgcacc ggaagcgtcg gaacagtatg 1020
cgcagagtcg aacgcaagta agaaagacgc accctcacac tcgcttactt cgagccatac 1080
aacggatcaa agctgcgcgt atctcggctt gtaagggccg gaaagcaacc tcggagatgg 1140
acacgtcaca tcaccaactt atcgatctcg gccgtcgacg tcgcagagag ggcgagagaa 1200
gcggtgaagg agggaaacaa cccctcgaga gcatgatccg accgaatctg cagcgcagga 1260
agccgttaca agcccgcctc gagcgcaggt cgggtccagc cgggggacga aacgcgcgag 1320
gctgattcgt gagcgaagga agccgcatcg acaagttcgc tcccctttgc cctctttccc 1380
atcacccgtt ctcgccttac ccgctcagaa caacaccaga tcactcacaa tgtc 1434
<210> 57
<211> 888
<212> DNA
<213> Aspergillus nidulans
<400> 57
ctgtacagtg accggtgact ctttctggca tgcggagaga cggacggacg cagagagaag 60
ggctgagtaa taagcgccac tgcgccagac agctctggcg gctctgaggt gcagtggatg 120
attattaatc cgggaccggc cgcccctccg ccccgaagtg gaaaggctgg tgtgcccctc 180
gttgaccaag aatctattgc atcatcggag aatatggagc ttcatcgaat caccggcagt 240
aagcgaagga gaatgtgaag ccaggggtgt atagccgtcg gcgaaatagc atgccattaa 300
cctaggtaca gaagtccaat tgcttccgat ctggtaaaag attcacgaga tagtaccttc 360
tccgaagtag gtagagcgag tacccggcgc gtaagctccc taattggccc atccggcatc 420
tgtagggcgt ccaaatatcg tgcctctcct gctttgcccg gtgtatgaaa ccggaaaggc 480
cgctcaggag ctggccagcg gcgcagaccg ggaacacaag ctggcagtcg acccatccgg 540
tgctctgcac tcgacctgct gaggtccctc agtccctggt aggcagcttt gccccgtctg 600
tccgcccggt gtgtcggcgg ggttgacaag gtcgttgcgt cagtccaaca tttgttgcca 660
tattttcctg ctctccccac cagctgctct tttcttttct ctttcttttc ccatcttcag 720
tatattcatc ttcccatcca agaaccttta tttcccctaa gtaagtactt tgctacatcc 780
atactccatc cttcccatcc cttattcctt tgaacctttc agttcgagct ttcccacttc 840
atcgcagctt gactaacagc taccccgctt gagcagacat caccatgg 888
<210> 58
<211> 617
<212> DNA
<213> Aspergillus niger
<400> 58
gtcgacgaga tcgtaggagt gagtacccgg cgtgatggag ggggagcacg ctcattggtc 60
cgtacggcag ctgccgaggg ggagcaggag atccaaatat cgtgagtctc ctgctttgcc 120
cggtgtatga aaccggaaag gactgctggg gaactgggga gcggcgcaag ccgggaatcc 180
cagctgacaa ttgacccatc ctcatgccgt ggcagagctt gaggtagctt ttgccccgtc 240
tgtctccccg gtgtgcgcat tcgactgggc gcggcatctg tgcctcctcc aggagcggag 300
gacccagtag taagtaggcc tgacctggtc gttgcgtcag tccagaggtt ccctccccta 360
ccctttttct acttcccctc ccccgccgct caacttttct ttccctttta ctttctctct 420
ctcttcctct tcatccatcc tctcttcatc acttccctct tcccttcatc caattcatct 480
tccaagtgag tcttcctccc catctgtccc tccatctttc ccatcatcat ctcccctccc 540
agctcctccc ctcctctcgt ctcctcacga agcttgacta accattaccc cgccacatag 600
acacatctaa accatgg 617
<210> 59
<211> 388
<212> DNA
<213> Ashibia gossipii
<400> 59
cactatacgt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac 60
cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat 120
ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc 180
gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac 240
agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt 300
gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca 360
catcacatcc gaacataaac aaccatgg 388
<210> 60
<211> 963
<212> DNA
<213> Rhodoturula grammis
<400> 60
ggcgcgccta cgtctacgtc aagggcaatg ccgacgtgac caaggccatc ggccaggacc 60
tcgccttctt ctcggtccct gtcgagctcg gcgtgcgtcc cgccgctctc tctctctctt 120
tctctcggcc gcgcctcacg tgatccacga cgtcgtactg acccttgcga atgtgcgcgc 180
ccgcagccca acggcgtcga gaaggtgcac ccgctcggcg acctgacggc gttcgagaag 240
gagctcctcg aggcgtgcct cggcgagctg cccgggtcca tctccaaggg cgagtcgttc 300
atccagggct ccaagctctg actcgccggc gcatcgacgg gcgcgagcca caaggcgagg 360
atgtgagagg aggcgtttcc tccaccttgg accccatctg ccgcctccct ttctctctct 420
ttctttccct tcctctctct ctctctctct ctcgttctcc tccttctggg cctctcggac 480
ctcttcctcg ccgtcgactc gtgaaaatgc agtgcgcgtt tctgtacctt gtcctgcgag 540
agagatctgg ttctgcgagg gtgagtcgtt gccttggccg tggcacgcct cgccgcagcg 600
agagagaaga ggccacggtc caggacgacg acgacgagga ggaagcgcaa aaggcgagac 660
accgagtgcc atcgattccc cgctcgaacc tgctcacggc tgtcgaaggc ggtgcgccac 720
ggtgcttgcg ggagcgaaag caagctggcg tcgtcctctt gaactggttc gagtccgtga 780
gggcggcgac gagaactcag gcgaggtgct cgcgtcggaa caagccgggc ttgtggtcga 840
gggagcgaga gcgaggcagc gccgtcgtcg ccgaggcaag agcggcatcg acaagttggc 900
ccgtcgcctc tcgctccctc ttctcctcct cccaccacca cctttctcca gctcgaacca 960
tgg 963
<210> 61
<211> 1009
<212> DNA
<213> Sporobolomyces roseus
<400> 61
ggcgcgccga agttatacct cagaggtctc aaaaacgaaa aagtcatgca agaatctcct 60
ttgacgtgag ggttatttct cttcctctag tagtctacga gaatcgcaaa gatcggaaaa 120
ctgatgcatc tttgtgttca cgggttagcg atttgatctt ttcgattccc aaaatcgtat 180
cgttcctgtc gcagggaact acgctcaaag ccggcactct gatcatcacg ggagtgagtt 240
ttgagctctc cctctatgag agtgcaaggt tcgtcgctga tggtgtaatc cgctcatgcc 300
ttcccctcta ccttctcctt tgtccattct ctctactacg gttgtcacat cttccttctc 360
cgacagaccc cgcacggaat tggagcgtac tcgaatcctc cggaattctt caaggacgga 420
gacgtcttca gggtcgagat ctcgggaggc atcgggagtt tggtcaacaa gatcgaatat 480
gaaaagtaga taatccgtta ctcaggtcaa tggtatggct tcgaagatgc tggaatcagc 540
cggaaagcaa agctggagag aaaaatcgag attgcgaaac gtgcgatgtc atttcgtttc 600
gagctcgcaa ccatctcgta tccctctgag ctacatacaa acgtcactac ggcctcggag 660
tgactccctg cgagcggatt gaaggagatc acggtcgaat cagctagacc ttcgcaacgt 720
tttcgcgctc gcacgttctt atcgatctac tgagattgac tcgaaaaagt cttctctcac 780
ggtcgattga actttgaatg aactctcagg ttgcgcgaga gccaatacga gccgaccaga 840
ggcaattcgg agcttcccgg aacgttccaa ggagagggat tttccgagag attacgattg 900
cgagatagaa aaaaggctag ctttcgattt cgagagagat tactttcaag ttcgctgctt 960
ccaactcttg ctccaacccc ctccactcct tctctacaaa acaccatgg 1009
<210> 62
<211> 2148
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<400> 62
ggcgcgccac catctcctcg tcgcttcttc cctctccttc ggcgcccaca ccgcttcgca 60
gggctcacgg actgctcaca tcgtttgtgt gcgtcgctgt gcatgtccac gcaccactcc 120
cagcccccac gagcgcctca aaagacgcgg acgcagacgc ccgccgaacg acggcacgcc 180
cctcttctca ctagcgcgac gaaccagctg cgacgattcg tgcgcttatg ttagccggac 240
ttctggcttg ctttgcgctg ctgcgtccgt cttgtggtgc ggatcggctc gatgggggtt 300
tgctcgtttg ctgggagacg gtcgcctctc cctcctcctc ttcactcctc gttagctttc 360
tacgctcatt ggttctgcga accatctaca tcacgctcgc tcgtcatgct cgtactacga 420
tcaacacccc tgctcgtcgt gctttccctc ctctccgtcc tctcggccgc gtccagcgac 480
ttgcccagcc aacttccccc gcacgccggt gagtctccca cacttccttg cgaccccaac 540
ccagcatctg acatccgcat cacgcagccc tcccgccttc ccactcctcc ctcttcaccg 600
actcctcctc ctcctcccct gattcctcgt ccctcaaagc cccgcagcct cttcccttca 660
aaatcaagcg cccccgctcg ctcgaacaag tgcagcagaa cctcgggaag aggctggcga 720
agcgcggcga ggaggggagt aagacggaga gggtgccgtt tggtcagagg agtgcgacgg 780
cggcgagtgc gggtggacaa ggtggagcgg ggacggggag ggcgacgcag cgcgttacgg 840
gcggaggaag cagaggtgca ggaggaggcg gagggagtgt cgcggctgct cagcctgtcc 900
cttcgactac ccagacggtc gagacaggct ctaagatcgt ctcgactggt cttctgaccg 960
tagcgtcgcc gtcgacggca gatggaggag gcgggacggt cacccaggtc gagacggcct 1020
cctcaggggt attgatcacc agcacggcgg gagcggcgag ttcagcggcg gcgtcggacg 1080
tcgctagcgc acaggcagcg gaggcgacgt cgagtacgag catgatcagc ggaggagcgg 1140
cggctggcgg gagtttaagc aggatgctgg cgggaggagt tgcgggtgca gccctgatcc 1200
tcctcgtgcg gtgagcaggc gaagcgagga gctcatgtag atacagcata gacagtatat 1260
atcgccagga tagcttgcaa cagccgccgg tcggtttatt ccattgtcct cgaccccatg 1320
cgaaggcgag ctctgctcgt cagctggcca agctggccag cagacgagcg ttggggtggc 1380
ggaacgccaa cggcatggag taaagcagcc gtgaggatga cggaggagct cgggcgaggt 1440
gatggggatt ctagcaggaa cagcagagcg gcgaggagga gaggaaccgg aagcacagtc 1500
tcgtggccgc ttgttgcaga tcccagtgtc gctagagtgc tcgtcgtcat cagagcgagt 1560
gaacaaagcg atgccctgaa gaacgatgag cgaatgagtc gaagcggcgt ctaccggtga 1620
actcggggtg tggcaaatga gcgagacgag gagtgcccgc cagagttgcc acgtcgaccc 1680
cacgtcggaa tcgacgttga tagagtgaac gaagccattg cagaccccag aaggtggcca 1740
tgttgtggaa gcgagggcag gagcgagggg agaaggcgag gaggaggagg ggctggggaa 1800
gcccgtccgg gaatggcgca gctgggtgcc ggggatgtgc gcgagtggcg gaggagtcga 1860
gcgtgagagt tctggaacac ggggcgcgca caagggtcga gggccgtgac gagttcgccg 1920
ggcggtggtc gggctgaggg cgagcgcgcg ttggggacga cgacgcccga cgccctcgct 1980
cttcgtcctc accgcttccc ggagaacttt gctgtactct gcttctccct tcacactctc 2040
acacccactc acacaccctt ccatccacac acaagctatc cgcacacctc tcacacccga 2100
ccccagctcg ccccatcctc ttcgcacccg gctcatcgaa aaccatgg 2148
<210> 63
<211> 1027
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (1022)..(1027)
<223> NcoI cloning site
<400> 63
ggtaccgtgc gagaagaagc gaggcacgcg aagcggtaga agcaatgaag cgaggcgaga 60
gcgagagagg cagggcttca gccatgtcca gctgatcggc tgtaacgtcg cgccgggcca 120
gtctgttgaa tttgttgcgt cgcctgagcg taatagaagt gcagtagtct actccgcatg 180
ccgagaacgt cgaagagcgc gaagtaggga gtcgagggaa gcgagggtgg caaacacagc 240
aacgacaagc ggttccgctt cgctcaaaag ctcgttgacg ttgttttgac gttttgaaga 300
cagtacaaca gcagcaagag gcgtgcgaag cgttggtggc gagagcagcg acaaggaggg 360
aggaatgagg gagtggtggc gagggctcgc aaacgggcgt acgcctcgaa tggagacgtg 420
cgagtcgttc ttcgacgtcc gagggatgcc gagcgccgag acggagcacg caacgagcga 480
gaggagagca gccgcgcaag gtgattcgag tggcgcaagc ggaggacgac gaggagacgg 540
acgagggagg aggagggatg gcgagcgagc atcggacggc ggggcgcgag agacggcgtg 600
aggagccggg tgtggagagt ttgaggaggc gcgggatgcg aagtggctgg gtgtgcggag 660
tgagcggtgg caaagagcgc acttagagtc tagagcgagg cagtagtagt agagctgtat 720
gaatgaatac aaagtgtgaa tacaacagtt tgtaatgcga ttctgagctt ggacgtgtgc 780
gcgcgagagg gcgacttgca agccagcgcc cgctcgctct tcttccttct gcacctcgcg 840
tcaaccctcg catctcacac ctacactcgc attcaaagtg cgtacactct cccacgacac 900
acggggacgg cgcacaccac cgcgcgtcgc ttgaacggcg tcgccacttc gagccgtcac 960
tgacttcgtc ctcgtcctcc ctcctctact ctcttgtact gtactgtgta ctggggggga 1020
accatgg 1027
<210> 64
<211> 956
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (951)..(956)
<223> NcoI cloning site
<400> 64
ggtacccatg ctgctgctgc ccctcaaagg tcctctcgtc cacgtccgac gagtctggac 60
agctttcaca gtcccgagag tgcaagagcg aggcgggctc acggttccgc aaaggagcgc 120
gaggtccgac cgccggccgg tctccttgcc cgcctcgcct cacctcctct tgcagcaggt 180
tcacctcttc gaggtcactc gatcgctcgc agcgatgcgc aggtacaagt acgctaggcg 240
agagcgtcga aagcggggtt ctgcgaggga ctggacgctg cagagcgcgg tcgagagagg 300
ctcgagtggc gctttgaccg ctcgacgcaa ggcatgcgct cctccgtttg agctcgcaga 360
tactgccgtg cgaagacgag cataggctgt ggctgcggta gcaaggagcc ggcgagagaa 420
agctgtgctc gagcaggacg agagacggtc cgcgcgcttg agaaggtcga ggtgaggcgt 480
cgcaaccggg ttggatctcg attctcggcg aactacggct tcggcgaggg ccaaagcgac 540
ggcaggccgc gcaagctggc caggcgagag cgcgagagtc gcgagctgaa gcgggcgcgg 600
ggtagagcaa gctggggaag cgagagaggg agagagagag agtgaggggg tggcgaggtg 660
gagacgaggc gagcggttgg cttgcgcgcg cgcgagaggg atcgaggcga gaggcgagcc 720
ccgagagtgg aaggaaggac gaggaaacct gcgtgcggag gcgccgcgcg cgcgtgccac 780
ctggctgagc acgggcccga gcttgaggga gctgggggcg cgcgagcgag acgagggcag 840
ggcgagcccg cgcgtggcgg ccgcctcgca acccaaggct cgccctggcc gccgctcttg 900
ctctctttcc tccaccttcg cgtctcacca ctcgaatctc acttcatcca ccatgg 956
<210> 65
<211> 1474
<212> DNA
<213> Rhodotorula graminis strain WP1
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (1469)..(1474)
<223> NcoI cloning site
<400> 65
ggtacccaaa gggagaggag cgggcggagg atggtggtgc cggacagggc gagagggaag 60
gtcgagggag agatgggagc gcgaaaggtc gaggcgggga gagggagggt ggggtcggga 120
ccaagggggc agagaggctc acaaggacgg aggagcttac tccgccttga ccttgcgggt 180
ggcggtggtg ccctcgcgga agctgcggcg cgaggaaggt cgtcagggcc gggcccaggg 240
gaggaacgag gacggcgacg acgacgcacc cgttcttgaa gcggcgcgag acgcccttca 300
ggtgctgcat gcggccagtg ccagtggtgt ggcggcgctt ggccttctgt ccccactcgt 360
ctgcgcggcg agggagaagg cgaggtgagc acgacggcgc gcgagggccg gacgaggctg 420
agagggggac gcacacgagc ggagcttggc cgcggggtag ccgcacgaag cgcacgactt 480
gtgctgcttg tggaacgagc ggcggccgca gcgacggcac agactgtggg accacgaggg 540
tcaaccgggt gctcgcgaga caggagcgcg gcttgtctcg aagcacgggc aaagagagcg 600
ttggacgcac gtgtgactct tggtgtggcg gagaccgaac gaggtggtac ccttcgtctg 660
tggggcgcaa ggaggagtgg gtcagcgtcg ggcctcgagg cgcctgggtc gtcgacctcg 720
cccgctcccg atcctcgcgc cgtcctgctc ctcctctctc caaccctgcg acgtgttgcg 780
gcagcagcag cttgctggga catgtgggga gggcggcaag gcgaggggag gtcgaggtgc 840
gaatgtgggt ggtcgcgctt ggcggggcag catgtcgtcg cggcctcgag ccgggcgggc 900
gacctggtgg ccgggtcgag cgagaggcgt gggagggagt ggcgcaaatg gcgtgcgctc 960
agaggcgggt tgtcgaggcg tcgaggcgga cgaggtcgag gaggtcgagg tgggaagctg 1020
ctgctgctgc tcgggcgtcg tcgccgcgtc ccgagtgccc cgtgcgcgcc cctgctgccg 1080
ctccttgggc cgtcctggtc ccacctgccc gtgccgtcct ccacgagagc gcgagtgggg 1140
ctgtgcgccg ggttgcgctc caactttgcg agagagcgag gacgggggca tggctcgctc 1200
gccggcctcg ggtcgttcga ggggtcgggg gcgggttgcg ggagggtggt gcgaggtggc 1260
gggcttacca ttgtcgcgtc ggagaggggg gtttggcctg cgagaagacg aggagacgag 1320
aggccggggg aggcgaggcg gcgaggcggc gagacggctc ggaccaagcg cgcgccgcca 1380
aagtctgcct cgccgctcgc gctcgcctcc ctcttgctct ccacctcctc ctaggaccac 1440
aaaggcaccc ttgtgtaggc gtaggtcacc atgg 1474
<210> 66
<211> 1327
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (1322)..(1327)
<223> BspHI cloning site
<400> 66
ggtacctgag cgggcgagcc gcgagcgagg agcgttgagg aggaagggag ggagggagga 60
gggagaggga gggaggggac ccccatcttt ctattcattc acaaagacga cggtgcggag 120
gggtccctcg agtgtttggg ctgggcttcg gagtctcgta gcgagcaagt agtgtttctc 180
tccgtttcga cagctcgtat tgtcatttct tgttcattgt cgtttccggc gactgcaggt 240
acgctgattt tcggcggaga cgacaagcac gtgggttgtg agcagcgagt tgagcaagaa 300
aaagcggacg aaggccctcg tcgggggctt caagtcaaga ttctgcggag attctgcgag 360
agactgcaag cgttgaacct gttgagatct cgtcggacga cagcacagtg tccgtctcgc 420
tcaatgcgat aggaagcgag agagaggagg aggatatcgg aggaaggcgt gtttgcgttc 480
gctccaggcg tcgcaagatc cggcgtagag cacaatcgtc gttggttcga cgtttgtagt 540
tcgtcacgag tgagggcgaa gcctggcaag caaagaaggg gacgagcgac tcggcagcta 600
tcgctggagg agggcgactt tgtggcccgt ttccgtcgag ctcgacgcga gtgagcgcag 660
ggtcggtccg aaccgatgcc atggacgcag tgagcgaggc cggatgtgcg atgctgtttc 720
aagcgagcga aggaagggag aaagcgagcg agaggtcctc ctcctgtctt cctcacgcct 780
tccgaaggcc gacaagaggc gtagacgtcg acgagtcaac ggtttgacgt cgctcaggcc 840
tgtagcgggt cgtcggaagc tgggaaagag aggaaccaac gagtaacaag cgcgagagtc 900
tcctcaaggc ggacaattgc ctcgcttcgg tcccggtcga gctcttccag taccagcgag 960
ggcgaaagtc gtcgatgcgt gcgcatccaa ggccaagcgt cgcagtcgag aagagcgaga 1020
gtgaagcgag tgaagcggga gagtgagagc gggtaatccg cgtacttacg agtgggttgt 1080
attccttctt gtaatggcag attacctcga ttggccacgt cacgttccgg gagtgcccgg 1140
gcgtgggcaa aagggcgagc gcggcgcctc tctctcttgc ttcctcagca gagcagctct 1200
cccctcgagt acgtcgacgg gctcactaca gctagcaaca gcaaggctac cacgccagct 1260
acacgccagc tcacccaact cacaccgctc gttgtcgccg cgcgccgcag gaaaactttg 1320
ttcagtc 1327
<210> 67
<211> 1509
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (1504)..(1509)
<223> NcoI cloning site
<400> 67
ggtaccgctc atcatcgagc gagggcagca gcgacctgcg gactggccga agaacttccc 60
cggccagcgc ttcgaggaca aggacattcg cacgcctcgc tctcagtggc ggtacatcaa 120
gctggcgacc acagacgacc tctcgccgac tgaggagaac acgacgtcct gcgccgtccg 180
gtacggcgag gactcgcagc tcgccatctt ccacgttccc ggcaaggggt acttctgcac 240
gcagcagatg tgtcctcgtg cgtcgcgcct gctctctctt tctttatttg ctggctgact 300
cgtgctgact cgcccgaaac ctcagacaag cgcgccttca tcctcgagca tggcatcgtc 360
tcggacgacg ggagtggcca cctctacgtc tcctgcccgc tccacaagcg caacttccgc 420
ctcgacaacg gcgactgcct caacgacgag gagtacaaga tcctcgcgtt tgatgtcaag 480
gaggagaacg gcgacttgct cgttcaggtt cctccgcctg acgagctcga cgctttgatt 540
gggtgcgtct cgcttagccc tctctcaaag acctgagctg acccttctga ttgtccgcag 600
ctcgtcgaag tggatggtgc gcaaagcgac cgccgaagcc ttcggtcgca acgcagcgac 660
agccatcgag tgcgtcccct ccaagcttct gttttccgcg cgcacactag gctgacgaca 720
agtctctgca ggatcgtcgg accgtcaggc gaggttgacg aggacaagaa ggcagcggga 780
acagagtgcg gcgaagcgga taagtcttgc gggacgcaca agctcgagtg gtgattcttg 840
cgggtccgtc acagccaatg tatctatctc tagatgtcct tctcgggtat atcagttgtt 900
cgtgcatcgt agacgtcgtt tagcagctct cgttcagcca cttgcgaagg cccgcttctt 960
cgacgacaag gacggcttcg cttcctttac ctcgtcgtct gagcgttctc aagggaccct 1020
cctacgccct tcttcgcaca ggagcggccg acgaggcagc cttgctggct tatcgtcgct 1080
tccgcctttc atgctcgagc aagtcctcct gcgagtgtcc cgacgtcggc ccgccttgcc 1140
caaggtcgcc gactgtccta tcgcgacact gcgaatgcac tgctgtccgc gccggagact 1200
gtgcggcgcg aattgagggc aaagtcgtgc atttgcgaaa cggtatccgc tcgaagggcc 1260
cacgatagac ctccaccggc ctcaaacttg gcgacagggt cgcttccgac ggcggacagc 1320
aagttaggct ttggcgtcgt cgctgcgatc cgctttgcgg gaccccttat cgcgactgcc 1380
ggattcgatt ggcgatatct ctcgctcgct ggcctcgctg gacagctgga cagtctctgc 1440
agcgtcgaag cgacgtcgat aaagtcagcg acgtcctcgc gaaccaagaa gaatcacccg 1500
ccgccatgg 1509
<210> 68
<211> 1606
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> SpeI cloning site
<220>
<221> misc_feature
<222> (1601)..(1606)
<223> NcoI cloning site
<400> 68
actagtcgcc agggaacgca gagaaggcgg gacgagcgag ggtgagtcgc gcaagtcgac 60
gaagaagccg aggtcaacgt cgagggttgt caagttctgg cagacggaca gcagccacac 120
taatcgctgc cgacttctga tgcatcgtcg cgcgggtgtc agctcctagc cgtggaagga 180
ggaagccagg acgttctcac cgggcgacgc attgcttgtg ccagacctcg aggcgagctt 240
gtcgcgtcga aagaccggcg acctcgtccg gctcaaattc cagcccgagt gattgccctg 300
acaacaaagg cgaaaagctg aagccggtac caaaggtcgg tccatcgaaa gtcgcgctcc 360
gaagactggc gtcgacggat ctgaccatcg ctgcccctcc tgcgtctgct ttgaggcacc 420
ttacagcctg ctcgtctcgt tcggagcctc cgcatccgct tggcaggacc acctcgcgac 480
cagtgacctc ccttgcgatg gctcgccaag tcttgcatac tccggcgacg ttgcggaagg 540
cgcaggtggg gcaggagatg cggagcgttg tgaactgtcc gttgacgagg agtgtcggcg 600
aaagaagggc ggtggtgaga gagtaggtga ggatcttgag gaggagttca ggaggaagtg 660
aggaaaggtc tgccggtgac tggtaaggct gaagcatgat ggcgagtgta gccaagtgat 720
ccgagcgacg atcaagagac gaaggacgag acaacgcttc agcgcgcgaa gagagcgagc 780
gaggaccctc ctggtcgaga ggctatccag tcgccaaccg gtacccatcc agtttgcagg 840
gttgaaacac agctgagagg atcagcgagt ggtagcgcaa actcctaagg cgctgaacgt 900
caaggacagc gagcgtgagc gtgtggaagc gacttgcgaa ggccaaactc gtgtcgcgct 960
ggccaaccgc cgtgccgctt tgacgcgctt ctgcgccctc cgcctattca gagagtatgc 1020
ttcgtcacgg cgtgggcgcc aacatcggcg caggagctgg cgggacggga agaaagccgc 1080
aaccgcggtc ctcgaccttc aacgtcccgg gaggcccgtc cacgactccc agacgtctct 1140
gcttgttgtt ctacgtcgtc gcggcgttgt gcagagtcca gcgcgcgccc gtcgtcgact 1200
tctgacaagc gataaattcc gagaccagcg ggagaaggcg gaacgagagg aggaggcgag 1260
ctggcgtcct tgcgacctcg ttgagcagtt caagcgagca gattgagcag cagtgcgtcg 1320
agtgagccaa ctcacgttct catatcggtc cctgagcgat atcgatgagg cgaaggacga 1380
cgacgagcga actgatctcg cgctctccct cttccccttc actctttcca ctcagaaaca 1440
acacgtgcgt cttctctgaa cgctatcaga caatccagga ccatcgctga ccgcgcgctc 1500
actcgtcgcc tgacttcatc gcccaaccaa cccgctcgtc accgactgga tctctccctc 1560
ccctcacacc acctgttgcg ctgcgatact cctctcacag ccatgg 1606
<210> 69
<211> 1408
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> SpeI cloning site
<220>
<221> misc_feature
<222> (1403)..(1408)
<223> BamHI cloning site
<400> 69
actagtcggc gaagaggagg ggtgttagga gggagagctg tgcgaggggg agaggtcagt 60
ccgagcacga tacgcgagca ggccaagcgg cttcatgttc actccaagct cgatgcggtc 120
gagaagtacg agctccttga cgagtgaaag ggagagaaga gagactcacc aaccccggcg 180
cgaccttcac gaaccgcaac ttcccctcac acgcctcact acacgccctc tcaaacacct 240
cgagcgtcgc gtagtgcgag agcttgaaga ataggtaggc gaggaaggag gaggcgaaga 300
gggcggccat ctagcgaaca agttgggtta gctgggtggg ggaaagggaa gaggggagga 360
agagggggag ggtaccatga tccagcctat gtcgacttgc agttggaggg gcatcctgtg 420
cgcgtactgg cgtcagcggc ggacgacaag gcaggtagac gagagagggg gccggggaac 480
gcactcgcag ttgcgcacgt ccgagaggaa catgtacgag tagccggccc acatgacgct 540
cagcagcgcc gcaagcgtgt agtggaagat gagcctgccc attggcagtc agcgccgacg 600
cggacaaact ctgggtgaag agaaaggaga gaacgcacca cttctcctcc ttcagcactc 660
ccctcgccca gaccgccccc aacacaatac agacgagatc gatagttgcc ccagtcacag 720
cgagcgcgag ttggctgtag acgaggtgtt tgagccgtct gtgggcggga gaggtgcgga 780
tgcgctggat ctgcgcgggg gagagttttg ggacgaagtt ggggtcgtcg gccatggtga 840
gcccgtcgta gtagctgtct gagcgagcct agtagtgcgc tggacgagca gagcccagag 900
tcgagacgag cgtgagcagg agacgaggtt cggagtgtcc gcggagggcg acgagacgac 960
gagcgagctt gggagaagcg cgagcatgtc cagcagcgta gtctcgaggc cgccagcagt 1020
agtagagcac agcaatgagg caggaaggag cgcaagggag ggaaagagcg cgacgaaggg 1080
tcgaggtgat gaagtccaag gacaggggga ccaccctcgc ccgcttctcc ctcgctctcc 1140
ccacgaagtg accacttgta aggctggtaa ttcattccat acagtctaca tacacttgca 1200
gccatccgct tcccctgcga tgccagtttc ggtcaccgtg ggactccgat gcgatgatgc 1260
ggccgagttg gcttcctcga cccgctctca cacgctcata ccagcctctc ccagcctgct 1320
accgctctct ggctctgcca aacacccact cgagcacacc cacccaacca gcgaactcgc 1380
ccagcctttg aaccgcaatg gcggatcc 1408
<210> 70
<211> 1480
<212> DNA
<213> Rhodotorula graminis strain WP1
<220>
<221> misc_feature
<222> (1)..(6)
<223> SpeI cloning site
<220>
<221> misc_feature
<222> (1475)..(1480)
<223> NcoI cloning site
<400> 70
actagtcgtg cagaaggaac ccgaggaggt cagtgcgcgg tcgagagagg aaaaggagag 60
agagacgcac cgagcgaggc attgcgcgtg ccagcgctca gcctgtacct cgcgactgct 120
gatgaaagcg tcgtcgccgc ccgacatgac cgccgcctcg ctcgactggt cgccgacgac 180
gtcgacgccg gaaggaggtg cgccccagcc gcaggcggcg cgcctcaggc tcacgtcgac 240
gtgccggacg ttcgacgctc gccacgggtc gtgctcgagg gctttgacga cgagctcgtc 300
gcgctcggcg ctgccgcacc cattggcgaa cgtgacgtcc tggccgagca cctcctgcgc 360
gatggcgcgc cacaggcggc agacggacgc gacctggcgg taggcgcacg tgcgacaggt 420
gaccgtgagg tgcgcaaact ggccgtggac gaggagaggc tcagagatga gcgaggtcgt 480
gagggcgtgg cggatgacct tgaggaggag ctcgtgcggc acggcagaga gggtcggcat 540
gatggtgtgc ggcgcggtcg gcagtctcga gagagatgtg tagaggaaga acgatgtcgc 600
cagatcggtc gagcaggagc cggtgcgagg cggctcgagg accgtcgcgg tcgaggaccg 660
gtcacggctg gacgatcgag gagacgcgcc cccgtcgagc gcagcggcca gacgcaagcg 720
agcacctttg aggctgtact ccaaaacccg gagcgccggc tcgggagccg tgtcctcgca 780
ggatcctcgg tcgacagcgc cgagtcggag agggccagcc gacctcgggc cgcccgacgc 840
ccggccgcag ctcctccggt ccgacctgca gctcatccca gcagatcgac tttgagagcg 900
aagcccccag gaagctgcct gagcgacctc gaggcttggg aaggtcgccg agccacggct 960
gggagagcga gctccctcac agtcgagacc ggctccaagt cgaatcgcac actcgtagct 1020
gcaccgcaaa agtgtgtgca gagctggagc gagcgaccgc gcgaggcgcg agggtcgcga 1080
gaaagcgggc gagcggtgcg agtgcgcccg agacgccgag agagggcgcg agggcgagcg 1140
ggcctcgcga gccctctgga gcgtgcagag gcggcgggga ggagcagagt gagggaggga 1200
agaccctcca gagctggcag gagccaacgg agcgcggaaa tcagtgagat cgatgcggtt 1260
ctcgagacga ttcgaccgcc ctcgtcgtca acgtcgcgcc ctcgtccctc tcctcttccc 1320
accacctctc cggtacctct acacgagtgc gttctgtccc gagatctgat ctcgacgccg 1380
cacggcactg actgaccgcc cacctcgtct ccctcgcccg tcccacactc tcccttccga 1440
cctcccacct cctcgctcaa cccctctcgc ctcgccatgg 1480
<210> 71
<211> 1532
<212> DNA
<213> Rhodotorula graminis strain WP1
<220>
<221> misc_feature
<222> (1)..(6)
<223> SpeI cloning site
<220>
<221> misc_feature
<222> (1527)..(1532)
<223> NcoI cloning site
<400> 71
actagtcggg aggattgatg atcgggttgc ggtacaaggc gaggccgagg acgagctgga 60
gggcgccgag gacgagggtc gagatgccga gctgcgaggg gacgaggccg ccaatcgagg 120
tcgtgtcaga cggccgtacc gagtgatctc atagcgggtc cgccgacgca gaaagacgaa 180
gagaaactca cgacgagggg gctcagcttg acaaagttga gctttgtcgt gcagccagac 240
gagctgcgat aggagcgagg gtcagtgcgc ccgctctcgg atgaatgcgc agtcgaggag 300
ggacaggggc gcaccaggtg acgatgaaca tgtcgaggac gccgtactgc agcatcttgc 360
ggaagaggta gaaatgccac gccgaagcga aggtgacggc aaactgcgag agggacgagg 420
tcagtgcgag ggtccgcaac agggagaggg cactcgtcac ggaccatcat ccagcccagg 480
tcgatctgta gctgatcagg gaagctgtgc cgggtgcgag agaggtcaat gtcgaagctt 540
ggcagctcgt cgaggaagaa gaggacggcg agggacgcac gccatgaact tgatgtgacg 600
ctcgaagacg accgagtagc cgagccacat cattgtgagg acgccggcgc agacgtagtg 660
gaagatgaac ctgagagggc aagaggtcag tctcgaaacg agggaggaag ccggctcgag 720
caggacgagg cgggcgcaac ggacgcacca tttctcggcg gcgacgatat tgcgagccca 780
cacggccccg agcactagac aggtgatgtt ggcgaccgcg ccgccgacgg cgagagcgag 840
aacgctgtag acgaagtgct tgatgcgtcg gtggattggc gctcgtcgga tcctcgcgat 900
ctgggccttg gtgagcggtg gtggcgggcc aagcggtggg ccagcagctg tgctcatcgc 960
agcagcggtg cggcgcaaga gcgactgtgg agctcgaggg agaggagcgc ggcaggggaa 1020
agcgagaccg aggaggagcg agcgcggaca ggcgaggcgg accggacgtt ccggtgcggc 1080
tcgactggcg tgcgagacga gcaggccgtc gccggaagca gccgtgtccg gcggaaagag 1140
ccaggcgcgc gagcggggcg gagcagacag cggcggtccg agcgcgcggg gcaggttcga 1200
cgaaagtcgg gctcgggtca ggctcgcgcg agcgcatgag atgccgtcga gcgagcccat 1260
gtacagagtc gagcgagaga gcgaagtgcg tggaaggaga gtggtccaag agtggagcgc 1320
cgtggagatg agacagatga tggcgaacct cggccacagc ctctcggtcc tgccacagca 1380
gctctgtgag tctccctgac ccgccagccc gcgcttcaga actcacagac cacctacaca 1440
gactcgcgca ccagctcgaa ccgcgccaga ccaccgcctc gccgcctccc cacctcgact 1500
gcttccgaac ctcacaagct cgaccaccat gg 1532
<210> 72
<211> 1222
<212> DNA
<213> Rhodotorula graminis strain WP1
<220>
<221> misc_feature
<222> (1271)..(1222)
<223> NcoI cloning site
<400> 72
ggtaccgcga ggggaagggc aggagagtcg ccggaacacc gggcaaggag acaaggacaa 60
agagcgagcg cgcacgcacc gtctggcggt cgggctgggt ggggcgggtc cgagtagcgc 120
tgaccgaaga gcgaggagga ggaaaagcga gcgcggcggg cgtgggaagg agggcaagga 180
gggcggcggc gacgccaaaa aaggccagat tcgcggggca gtgatcgagt gccgtccgcg 240
agctcaacca gcgagcgctc tctcgcgcgc agcggtgcgc ttctttcgcc agccgatgcg 300
caccgttcag aagcacgtcg cccgcaccga gagcgcctcc tcgcgagcct gtgaccacct 360
cgtcgacccg cttcccgcgg cttttctcgc cggcctggac cgccgctatc agatcgtgcc 420
catgagacaa gcgactcgtc gaaaggacga cgatctcgta gtactgggtc cctgcgcaac 480
gctcagccgt ccgtccccgt caaagtgctt cggcggaggg gaccgtgcgc gagacgccca 540
agttggcctc ctcaagtcgg tagatccagc ttaacgctat caagggttgc atggtgtagt 600
tggtcatcac gtcagtttaa cattcagttc actgaaggtc ctcagttcaa acctgggtgc 660
gatcaccttt ttggctcggc ggcattgcgc ccttacaccc gcacgggtct acttcccttt 720
gcaagcgacc aagcgaagca tcctctcgct cgtaaagctg ccggcgagga ggtcagacgg 780
gttggcgggc cgtcgaaggt cggctcaccc tcaacgctgc cggctgacca cgccaggcga 840
gctatcattg ctttgaaagc ttcgaaaacg cccaggcatg cacagaaagc cgcccgcgag 900
aggctcaagt tggcgccgag ctgcggtcga gagacgacga cgacgtggga gctccctcgc 960
ctctcctcct ttctctccca ccccatcagc ccaagtgagt cgctcgctct tccgcaaggg 1020
tcagcgcacg cgttgctccg cgacagggca gcgcgtgcgc tcaccagggt cccccgttcg 1080
cccggcgagt tggcactgac gaggtgcctt gcccccctcc gctcccctcc cctttggcct 1140
cctctctcgc acgcacactc tctccctgca ccccttgcac cttcccgaca ctctcccccc 1200
ccttcccacc gtccgaccat gg 1222
<210> 73
<211> 1507
<212> DNA
<213> Rhodotorula graminis strain WP1
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (1502)..(1507)
<223> NcoI cloning site
<400> 73
ggtaccacta cctcgtcacg acccagggtg cgttcccctt ctcgccgcgt ccacagccac 60
gacgttgcga gtccctaacg cgtccgctcc cccgtgcagg tggtcctccg cacggcgcct 120
acacgcagac atcgtcgacg ccgtcgcacg acgcccactt tgccgtccag accctgtcgc 180
gcttcggcct cgcgtacctc ctcgcgtcgt ccaacacgct caaggacacg tggctgagcg 240
tgtgcgcgcc cgccggcgcc aagggccccg aacccgacgt cgacgacctc gagctcgaga 300
agcgcgagca ccgcgacaag tggctcctcg ggcgcatcat gggccagggc aagcaggact 360
cggcgctcgg ggacgcagtc gctgtcgtga gtcctctact ctcggccgtt ctcgagactt 420
ggggtgcgag attgaccttg cgctcccgcc tctcgcttgc agcaattcca caagcacttt 480
ccgcacctgc gctcggcgca cctcttcccc ggctttgtct ttacgtgcgt ccctctcgtt 540
cccccctctc tccacgtgcg ccaccagcct gactcgcccc tcgcctgccc cgtcccgcag 600
caacgccctc gcgtcgacct cgctcgtccc ctcgccgatc ctgtcgctgt acaacctcgt 660
cgggcccctc gcggcgcgca tcctgccctt tggcaacctg cccgagacgt acgccgacgt 720
gcccgtgtac gtcgcggcca acccggcagc gcgcagccaa gggctcgagt actgcaacga 780
gcgcatgaag ccgctcggga gcccggcgtg ggccgagggc gcgacgggcg caaaggtgtg 840
ggacgggctg agggccatga tcgaggagtg agctggtggg cgggcgagcg aggagccgga 900
gaggaggggc ggaacgtgtt tgagaaggtc gcgctttgct cgtcggtcgc gggcgcagcc 960
gtggctgtag ccagtctcgc tttgcagtgt cactcttgta catagctgag caaggcctag 1020
cgtcgcgaga gagctgcgct gtggcgcctg gtcgaggccc gagagcgtcg cgctcagggg 1080
cgagctgctc gcggctcacc aaggggctcg agcggtgcgc gctcgacagg ggaccgagag 1140
ctgcaggaga cagaccggag gaaaaagctc tggcgagcga ggagcggggc cacactgagt 1200
ctggggaagc gacggacgag gatgagcgca tccactcttg agtttcgccg aggcgcgagc 1260
tggcggtcga caaccgagca agctcctcct cttcctccac cacactcgcc cctagcacac 1320
gtgagtctcg ctccctcgcc actgtcgacc agcacacgct cgtccaccgc cctgtgcgcc 1380
ctgtgcggct tgcggtcgag cgaggccgcg ggtcgggtct ctgccacccg aggaaccatc 1440
gatgtcgctg acgcttcgct cctcgtcctc ctcctcctcc cacccgccgc agctacctac 1500
accatgg 1507
<210> 74
<211> 1646
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> AscI cloning site
<220>
<221> misc_feature
<222> (1641)..(1646)
<223> NcoI cloning site
<400> 74
ggcgcgcctg aagctgtaca tcgaggtgga cgacagcgcg ggcaaggatg cgccagcaat 60
cgtttgtggg tcgcttcttt cctcgcagca cgcttttgtc ggctccctga tcagcacaca 120
agctaactaa cgctctggtt tcgctggcag tcatgcacgg ccttggctcg tcaacctcgt 180
tctgggaagc gcccttctcc cgctcaaacc tgtcctcccg cttccgcctc atccgctacg 240
acttcgacgg ccacggtctc tcgcccgtct cgtccctcga cgcagcagat gacggcgcca 300
tgatcccgct cgacgacctc gtcggggact tggcggctgt gatcgagtgg gctggggtgg 360
agaaggttgc gggagttgtt ggacactcga tgagcgggct ggtggcgagc acatttgcgg 420
ccaagtaccc gcagaagctc gacaagctcg gtgagtcgca ttgaaccttc ctccgccgtc 480
tcttctccgc tgacgattcg tcgacttggc cctgcttctc gcgcagtcct cctcggcgca 540
atgcgctctc tgaaccctac cgtccaaagc aacatgctca agcgagccga tacagtcctc 600
gaatccggcc tctcagcaat cgtcgcacaa gtcgtctccg ccgctttgtc cgacaagtca 660
aagcaggact cgcccctctc ggcagcgatg gtgcgaacgc tcgtgcttgg aacggacccg 720
agagggtacg cggcggcgtg tagggcgctt gcgggtgcga aggacccgga ttactcgagc 780
atcaaggccg agacgttggg tgcgttcgct tgttctcctt cctctgcttt tctcccagca 840
actgacgcaa gcgtctgcaa cacagtcgtc gcaggcgagt ttgactacct ctcgaacaag 900
gagacgaccg acgcgctggt caacgacatc ccgggcgcgg agaaggtcca gatggacagt 960
gtcggccact ggcacgccgt cgaggacccc gttggactcg ccaagatcct cgatgggttc 1020
ttcttgcagg ggaaatgagg ttgggaaggg gggatagact ggggagaacg gcaggtgcgt 1080
acgcagcgga cgtcggtcgg gaggactttt tcggggagga tattcgctga ctgactccga 1140
cgtcgctttc ctccttgcag tatcttcaga agggatggga ggaggcgaac tgcaagggta 1200
atgaacgaga caacgccgag ggaggaagcg ccggaactct cgggggcgaa gaaggagtgg 1260
tgtcttcgcc agcgaacagc ttccggggtg ggttggacag cgccagtaga attccagcgt 1320
cgcaacagag ctctagtcga ccgcgatcac ccacaaggac gagagcgggt cgcgccttgt 1380
ccgcttcccc atcctcgtcc tgctcttgct ctcttcccta ccacactctc ccgcttgcgg 1440
gctctctttc tcgcttggcg ctcctgctac cgctactcta gactctccta gtctccctgc 1500
acaaccatcc ctatcccctc cgcctctctc gcacaccccc cacagcttcg ttccccaact 1560
tcacttccga tgccgtgcgt cgcctccctt tcgcctggcg ggcccgcgcc tgcttccgag 1620
gacaactact gattgtggga ccatgg 1646
<210> 75
<211> 1450
<212> DNA
<213> Rhodosporidium toruloides
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (1445)..(1450)
<223> NcoI cloning site
<400> 75
ggtaccggcg cgctagtctg cagaactacg ccctctcaca cccaacttcc gactcgaccg 60
gcggtacgag cacgacctac ttctactgcc tgccatcgac atccgggcgg gtcgctgcct 120
accctgtgcg ttctgcgccc tccctcgtct cgggaggcag tgtctgacag aagctttgcg 180
cgcagtaccc cgtcaagatg caactctacg caacgttcgg cacagaagtc gccaagctcc 240
gcgcatcgcc gcctcaagct ctcgcgctgc ccgacggtgt cgtctattac gaggcggaga 300
agctcgagtt gccggctttg ccagcggcgg tcaaggttga ggtggagacg gagaaggcgg 360
gagtagcggg ggaggacaat gaggcgaagg gtgagatggt gctggtggag actcttacgg 420
tggagcagga ggagattgaa ttgggctcgg gagtcgtgca gattgaggag tcgttgctcg 480
tcaagctgga ggtcagcggc tgatccttcc gttcgttgca aggatcgtct gcatgtttcg 540
cttctctcaa tgacacaacc tggagagcgc tcccgtcagc gagaatcgag gacattccgc 600
agctcgtgag caagcggagg tgcgaggctc cctcgaaagc tgcgcctctt cagacggctt 660
gttctctcct gctctggtgg gctggcctga catgtaatgt gctccgccgc aagtccgtcg 720
tcggtctcaa ttcgacgttg aaagggcata gcgcaaggaa gaaccctctg cggacatgca 780
gaattactgg ctcgcctgct ccttcgtcta ctggaataag tcctgtctcg ttaaagcccc 840
aacgtcgttt ttcgacgttt gtaaggcgca agaggtgcta tgggctacgc aggaagctga 900
gaggacatag aagtcggggg aggaacggcg cagagcggca gttgcggaag catgaggaaa 960
gcgagacggt ccagcatctg cagcgccaat ccgcaatctc ctggttgagc ctgcaccgga 1020
agcgtcggaa cagtatgcgc agagtcgaac gcaagtaaga aagacgcacc ctcacactcg 1080
cttacttcga gccatacaac ggatcaaagc tgcgcgtatc tcggcttgta agggccggaa 1140
agcaacctcg gagatggaca cgtcacatca ccaacttatc gatctcggcc gtcgacgtcg 1200
cagagagggc gagagaagcg gtgaaggagg gaaacaaccc ctcgagagca tgatccgacc 1260
gaatctgcag cgcaggaagc cgttacaagc ccgcctcgag cgcaggtcgg gtccagccgg 1320
gggacgaaac gcgcgaggct gattcgtgag cgaaggaagc cgcatcgaca agttcgctcc 1380
cctttgccct ctttcccatc acccgttctc gccttacccg ctcagaacaa caccagatca 1440
ctcaccatgg 1450
<210> 76
<211> 672
<212> DNA
<213> Ustilago maydis
<220>
<221> misc_feature
<222> (1)..(6)
<223> KpnI cloning site
<220>
<221> misc_feature
<222> (667)..(672)
<223> NcoI cloning site
<400> 76
ggtaccggcg cgctagtcct taaaactgaa ggcgggaaac gacaatctga tccaagctca 60
agctaagctc tagtgattag atcttgctga taggcaggtt tgcttggaga atggggggaa 120
aagactgacc gaagaaacag cgagatctag aagtgataag cggaaagaat ctgacttgct 180
gtgatcagca gccaattttt ttttcgtttt ttttttttca ctccacatcg tcgtgcgtgc 240
acggtctgca tgtgtaaatt gtattcatcg aaagccacag ttgaatacat cagcccgatg 300
tggatttcga aaaccaatta atcttggaat tcacgcgctc agatcagtcc atagagtcga 360
cttcggctgt ttccaagagc ttcttctctg cgaggtggtt gcccgtgttt ctcgctggga 420
aaaaaggatc gattattatt cgcttctacc tcgctcgcac ccttggcctg ctgaaggaaa 480
cagcgccgag actcggtcac ggttgctggg ctccgtgttg atgctgggac ggcgcaaagt 540
ggggcccgcg cactcttcga gccaaggacc tcactcttca agaacaagcg ctgtcgccat 600
cgtcttcttc tttctgctcc accatcgaat ctttctttct cgtttcgaaa ccaaaacact 660
cttccaccat gg 672
<210> 77
<211> 963
<212> DNA
<213> Rhodotorula graminis strain WP1
<220>
<221> misc_feature
<222> (1)..(6)
<223> AscI cloning site
<220>
<221> misc_feature
<222> (958)..(963)
<223> NcoI cloning site
<400> 77
ggcgcgccta cgtctacgtc aagggcaatg ccgacgtgac caaggccatc ggccaggacc 60
tcgccttctt ctcggtccct gtcgagctcg gcgtgcgtcc cgccgctctc tctctctctt 120
tctctcggcc gcgcctcacg tgatccacga cgtcgtactg acccttgcga atgtgcgcgc 180
ccgcagccca acggcgtcga gaaggtgcac ccgctcggcg acctgacggc gttcgagaag 240
gagctcctcg aggcgtgcct cggcgagctg cccgggtcca tctccaaggg cgagtcgttc 300
atccagggct ccaagctctg actcgccggc gcatcgacgg gcgcgagcca caaggcgagg 360
atgtgagagg aggcgtttcc tccaccttgg accccatctg ccgcctccct ttctctctct 420
ttctttccct tcctctctct ctctctctct ctcgttctcc tccttctggg cctctcggac 480
ctcttcctcg ccgtcgactc gtgaaaatgc agtgcgcgtt tctgtacctt gtcctgcgag 540
agagatctgg ttctgcgagg gtgagtcgtt gccttggccg tggcacgcct cgccgcagcg 600
agagagaaga ggccacggtc caggacgacg acgacgagga ggaagcgcaa aaggcgagac 660
accgagtgcc atcgattccc cgctcgaacc tgctcacggc tgtcgaaggc ggtgcgccac 720
ggtgcttgcg ggagcgaaag caagctggcg tcgtcctctt gaactggttc gagtccgtga 780
gggcggcgac gagaactcag gcgaggtgct cgcgtcggaa caagccgggc ttgtggtcga 840
gggagcgaga gcgaggcagc gccgtcgtcg ccgaggcaag agcggcatcg acaagttggc 900
ccgtcgcctc tcgctccctc ttctcctcct cccaccacca cctttctcca gctcgaacca 960
tgg 963
<210> 78
<211> 2148
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> misc_feature
<222> (1)..(6)
<223> AscI cloning site
<220>
<221> misc_feature
<222> (2143)..(2148)
<223> NcoI cloning site
<400> 78
ggcgcgccac catctcctcg tcgcttcttc cctctccttc ggcgcccaca ccgcttcgca 60
gggctcacgg actgctcaca tcgtttgtgt gcgtcgctgt gcatgtccac gcaccactcc 120
cagcccccac gagcgcctca aaagacgcgg acgcagacgc ccgccgaacg acggcacgcc 180
cctcttctca ctagcgcgac gaaccagctg cgacgattcg tgcgcttatg ttagccggac 240
ttctggcttg ctttgcgctg ctgcgtccgt cttgtggtgc ggatcggctc gatgggggtt 300
tgctcgtttg ctgggagacg gtcgcctctc cctcctcctc ttcactcctc gttagctttc 360
tacgctcatt ggttctgcga accatctaca tcacgctcgc tcgtcatgct cgtactacga 420
tcaacacccc tgctcgtcgt gctttccctc ctctccgtcc tctcggccgc gtccagcgac 480
ttgcccagcc aacttccccc gcacgccggt gagtctccca cacttccttg cgaccccaac 540
ccagcatctg acatccgcat cacgcagccc tcccgccttc ccactcctcc ctcttcaccg 600
actcctcctc ctcctcccct gattcctcgt ccctcaaagc cccgcagcct cttcccttca 660
aaatcaagcg cccccgctcg ctcgaacaag tgcagcagaa cctcgggaag aggctggcga 720
agcgcggcga ggaggggagt aagacggaga gggtgccgtt tggtcagagg agtgcgacgg 780
cggcgagtgc gggtggacaa ggtggagcgg ggacggggag ggcgacgcag cgcgttacgg 840
gcggaggaag cagaggtgca ggaggaggcg gagggagtgt cgcggctgct cagcctgtcc 900
cttcgactac ccagacggtc gagacaggct ctaagatcgt ctcgactggt cttctgaccg 960
tagcgtcgcc gtcgacggca gatggaggag gcgggacggt cacccaggtc gagacggcct 1020
cctcaggggt attgatcacc agcacggcgg gagcggcgag ttcagcggcg gcgtcggacg 1080
tcgctagcgc acaggcagcg gaggcgacgt cgagtacgag catgatcagc ggaggagcgg 1140
cggctggcgg gagtttaagc aggatgctgg cgggaggagt tgcgggtgca gccctgatcc 1200
tcctcgtgcg gtgagcaggc gaagcgagga gctcatgtag atacagcata gacagtatat 1260
atcgccagga tagcttgcaa cagccgccgg tcggtttatt ccattgtcct cgaccccatg 1320
cgaaggcgag ctctgctcgt cagctggcca agctggccag cagacgagcg ttggggtggc 1380
ggaacgccaa cggcatggag taaagcagcc gtgaggatga cggaggagct cgggcgaggt 1440
gatggggatt ctagcaggaa cagcagagcg gcgaggagga gaggaaccgg aagcacagtc 1500
tcgtggccgc ttgttgcaga tcccagtgtc gctagagtgc tcgtcgtcat cagagcgagt 1560
gaacaaagcg atgccctgaa gaacgatgag cgaatgagtc gaagcggcgt ctaccggtga 1620
actcggggtg tggcaaatga gcgagacgag gagtgcccgc cagagttgcc acgtcgaccc 1680
cacgtcggaa tcgacgttga tagagtgaac gaagccattg cagaccccag aaggtggcca 1740
tgttgtggaa gcgagggcag gagcgagggg agaaggcgag gaggaggagg ggctggggaa 1800
gcccgtccgg gaatggcgca gctgggtgcc ggggatgtgc gcgagtggcg gaggagtcga 1860
gcgtgagagt tctggaacac ggggcgcgca caagggtcga gggccgtgac gagttcgccg 1920
ggcggtggtc gggctgaggg cgagcgcgcg ttggggacga cgacgcccga cgccctcgct 1980
cttcgtcctc accgcttccc ggagaacttt gctgtactct gcttctccct tcacactctc 2040
acacccactc acacaccctt ccatccacac acaagctatc cgcacacctc tcacacccga 2100
ccccagctcg ccccatcctc ttcgcacccg gctcatcgaa aaccatgg 2148
<210> 79
<211> 1009
<212> DNA
<213> Sporobolomyces roseus
<220>
<221> misc_feature
<222> (1)..(6)
<223> AscI cloning site
<220>
<221> misc_feature
<222> (1004)..(1009)
<223> NcoI cloning site
<400> 79
ggcgcgccga agttatacct cagaggtctc aaaaacgaaa aagtcatgca agaatctcct 60
ttgacgtgag ggttatttct cttcctctag tagtctacga gaatcgcaaa gatcggaaaa 120
ctgatgcatc tttgtgttca cgggttagcg atttgatctt ttcgattccc aaaatcgtat 180
cgttcctgtc gcagggaact acgctcaaag ccggcactct gatcatcacg ggagtgagtt 240
ttgagctctc cctctatgag agtgcaaggt tcgtcgctga tggtgtaatc cgctcatgcc 300
ttcccctcta ccttctcctt tgtccattct ctctactacg gttgtcacat cttccttctc 360
cgacagaccc cgcacggaat tggagcgtac tcgaatcctc cggaattctt caaggacgga 420
gacgtcttca gggtcgagat ctcgggaggc atcgggagtt tggtcaacaa gatcgaatat 480
gaaaagtaga taatccgtta ctcaggtcaa tggtatggct tcgaagatgc tggaatcagc 540
cggaaagcaa agctggagag aaaaatcgag attgcgaaac gtgcgatgtc atttcgtttc 600
gagctcgcaa ccatctcgta tccctctgag ctacatacaa acgtcactac ggcctcggag 660
tgactccctg cgagcggatt gaaggagatc acggtcgaat cagctagacc ttcgcaacgt 720
tttcgcgctc gcacgttctt atcgatctac tgagattgac tcgaaaaagt cttctctcac 780
ggtcgattga actttgaatg aactctcagg ttgcgcgaga gccaatacga gccgaccaga 840
ggcaattcgg agcttcccgg aacgttccaa ggagagggat tttccgagag attacgattg 900
cgagatagaa aaaaggctag ctttcgattt cgagagagat tactttcaag ttcgctgctt 960
ccaactcttg ctccaacccc ctccactcct tctctacaaa acaccatgg 1009
<210> 80
<211> 1858
<212> DNA
<213> Rhodosporidium toruloides
<400> 80
atgggccagc aggcgacgct cgaggagctg tacacacgct cagaaatctc caagatcaag 60
caagtcgagc cagctcttct cctcaccacc ccacaacata ccccgcagcc cacgacagct 120
ctcccacagc acccacagcc tgctgaccag ctcgagtgca tccacagatt tgcgcccttt 180
ggcgtcccgc ggtcgcgccg gctgcagacg ttctccgtct ttgcctggac gacggcactg 240
cccatcctac tcggcgtctt cttcctgctc tggtgcgtca ggcttggcgt ggattgggag 300
tagcgggcga ctcagctgac ttgcgcatcc gctccagctc gttcccaccg ctctggccgg 360
ccgtcatcgc ctacctcacc tgggtctttt tcattgacca ggcgccgact cacggtggac 420
gggcgcagtc ttggctgcgg aagagtcgga tatgggtctg gtttgcagga tattatcccg 480
tcaggtgcgt cgtcccgtct gttgcgcgtc ttgcgacctc gctcacggcc aactcgcccg 540
accggctacc tccgaacttc ccgccaacag cttgatcaag gttcgtccac ctttccttca 600
gcttgagtga tctgtagagg agctgcagga tcaagcccaa cccggggagg acctcggagg 660
acgacgccgc tgacttgctc tcctcctaca gagcgccgac ttgccgcctg accggaagta 720
cgtcttcggc tatcatccgc acggcgtcat aggcatgggc gccatcgcca acttcgcgac 780
cgacgcaacc ggcttctcga cactcttccc cggcttgaac cctcacctcc tcaccctcca 840
aagcaacttc aagctcccgc tctatcgcga gttgctgctc gccctcggca tctgctccgt 900
ctcgatgaag agctgccaga acatcctgcg gcaaggtgcg ccagtcattc cgaacgggcg 960
gtcgagcgtg aactctgggg atgggaagag ctgaccttct gcctcactcc atccatgcag 1020
gtcctggctc ggctctcacc atcgttgtcg gtggcgcagc cgagagcttg agtgcgcatc 1080
ccggaaccgc cgacctcacg ctcaagcgac gaaaaggatt catcaagctc gcgatccggc 1140
aaggcgccga cctcgtgccc gtcttttcgt tcggcgagaa cgacgtgcgt cctctgctcg 1200
acttccgcta gcgaagccct tcgctgacgc tcccggtttc ttcccccaga tcttcggcca 1260
gctgcgaaat gagcgaggga cgcggctgta caagttgcag aagcgtttcc agggcgtatt 1320
cggcttcact ctccgtacgt tgcgccgtgt cgcttcaatc tgtcgagcgt ccagtcgctc 1380
acgcagctac aactcccaca gctctcttct acggtcgggg actcttcaac tgtacgcccg 1440
agtctacgtg actagtctac cgtgggaggc actgaagagc acggctgacg tcccacctct 1500
ccgcgcagat aacgttggct tgatgccgta ccgccatccg atcgtctcgg tcggtgcgtc 1560
ccccctcgtc cctcctgacc tgcgggcttc agctaacaat tctcgacgac atctagtcgg 1620
tcgaccaatc tcggtgcagc agaaggacca cccaacgaca gcggatctcg aagaagtcca 1680
ggcgcggtat atcgcagaac tcaagcggtg cgttccagac gtctaccttt gcccgttgtc 1740
tcagactcgg taagacagat cactgacgct tcggtcactg gccgcgcagc atctgggaag 1800
actacaagga cgcctacgcc aaaagtcgca cgcgggagct caatattatc gcctgacc 1858
<210> 81
<211> 1047
<212> DNA
<213> Rhodosporidium toruloides
<220>
<221> CDS
<222> (1)..(1047)
<400> 81
atg ggc cag cag gcg acg ccc gag gag cta tac aca cgc tca gag atc 48
Met Gly Gln Gln Ala Thr Pro Glu Glu Leu Tyr Thr Arg Ser Glu Ile
1 5 10 15
tcc aag atc aaa ttc gca ccc ttt ggc gtc ccg cgg tcg cgc cgg ctg 96
Ser Lys Ile Lys Phe Ala Pro Phe Gly Val Pro Arg Ser Arg Arg Leu
20 25 30
cag acc ttc tcc gtc ttt gcc tgg acg acg gca ctg ccc atc cta ctc 144
Gln Thr Phe Ser Val Phe Ala Trp Thr Thr Ala Leu Pro Ile Leu Leu
35 40 45
ggc gtc ttc ttc ctc ctc tgc tcg ttc cca ccg ctc tgg ccg gct gtc 192
Gly Val Phe Phe Leu Leu Cys Ser Phe Pro Pro Leu Trp Pro Ala Val
50 55 60
att gcc tac ctc acc tgg gtc ttt ttc att gac cag gcg ccg att cac 240
Ile Ala Tyr Leu Thr Trp Val Phe Phe Ile Asp Gln Ala Pro Ile His
65 70 75 80
ggt gga cgg gcg cag tct tgg ctg cgg aag agt cgg ata tgg gtc tgg 288
Gly Gly Arg Ala Gln Ser Trp Leu Arg Lys Ser Arg Ile Trp Val Trp
85 90 95
ttt gca gga tac tat ccc gtc agc ttg atc aag agc gcc gac ttg ccg 336
Phe Ala Gly Tyr Tyr Pro Val Ser Leu Ile Lys Ser Ala Asp Leu Pro
100 105 110
cct gac cgg aag tac gtc ttt ggc tac cac ccg cac ggc gtc ata ggc 384
Pro Asp Arg Lys Tyr Val Phe Gly Tyr His Pro His Gly Val Ile Gly
115 120 125
atg ggc gcc atc gcc aac ttc gcg acc gac gca acc ggc ttc tcg aca 432
Met Gly Ala Ile Ala Asn Phe Ala Thr Asp Ala Thr Gly Phe Ser Thr
130 135 140
ctc ttc ccc ggc ttg aac cct cac ctc ctc acc ctc caa agc aac ttc 480
Leu Phe Pro Gly Leu Asn Pro His Leu Leu Thr Leu Gln Ser Asn Phe
145 150 155 160
aag ctc ccg ctc tac cgc gag ttg ctg ctc gct ctc ggc ata tgc tcc 528
Lys Leu Pro Leu Tyr Arg Glu Leu Leu Leu Ala Leu Gly Ile Cys Ser
165 170 175
gtc tcg atg aag agc tgt cag aac att ctg cga caa ggt cct ggc tcg 576
Val Ser Met Lys Ser Cys Gln Asn Ile Leu Arg Gln Gly Pro Gly Ser
180 185 190
gct ctc act atc gtc gtc ggt ggc gcc gcc gag agc ttg agt gcg cat 624
Ala Leu Thr Ile Val Val Gly Gly Ala Ala Glu Ser Leu Ser Ala His
195 200 205
ccc gga acc gcc gat ctt acg ctc aag cga cga aaa ggc ttc atc aaa 672
Pro Gly Thr Ala Asp Leu Thr Leu Lys Arg Arg Lys Gly Phe Ile Lys
210 215 220
ctc gcg atc cgg caa ggc gcc gac ctt gtg ccc gtc ttt tcg ttc ggc 720
Leu Ala Ile Arg Gln Gly Ala Asp Leu Val Pro Val Phe Ser Phe Gly
225 230 235 240
gag aac gac atc ttt ggc cag ctg cga aac gag cga gga acg cgg ctg 768
Glu Asn Asp Ile Phe Gly Gln Leu Arg Asn Glu Arg Gly Thr Arg Leu
245 250 255
tac aag ttg cag aag cgt ttc caa ggc gtg ttt ggc ttc acc ctc cct 816
Tyr Lys Leu Gln Lys Arg Phe Gln Gly Val Phe Gly Phe Thr Leu Pro
260 265 270
ctc ttc tac ggc cgg gga ctc ttc aac tac aac gtc gga ttg atg ccg 864
Leu Phe Tyr Gly Arg Gly Leu Phe Asn Tyr Asn Val Gly Leu Met Pro
275 280 285
tat cgc cat ccg atc gtc tct gtc gtc ggt cga cca atc tcg gta gag 912
Tyr Arg His Pro Ile Val Ser Val Val Gly Arg Pro Ile Ser Val Glu
290 295 300
cag aag gac cac ccg acc acg gcg gac ctc gaa gaa gtt cag gcg cgg 960
Gln Lys Asp His Pro Thr Thr Ala Asp Leu Glu Glu Val Gln Ala Arg
305 310 315 320
tat atc gca gaa ctc aag cgc atc tgg gaa gaa tac aag gac gcc tac 1008
Tyr Ile Ala Glu Leu Lys Arg Ile Trp Glu Glu Tyr Lys Asp Ala Tyr
325 330 335
gcc aaa agt cgc acg cgg gag ctc aat att atc gcc tga 1047
Ala Lys Ser Arg Thr Arg Glu Leu Asn Ile Ile Ala
340 345
<210> 82
<211> 348
<212> PRT
<213> Rhodosporidium toruloides
<400> 82
Met Gly Gln Gln Ala Thr Pro Glu Glu Leu Tyr Thr Arg Ser Glu Ile
1 5 10 15
Ser Lys Ile Lys Phe Ala Pro Phe Gly Val Pro Arg Ser Arg Arg Leu
20 25 30
Gln Thr Phe Ser Val Phe Ala Trp Thr Thr Ala Leu Pro Ile Leu Leu
35 40 45
Gly Val Phe Phe Leu Leu Cys Ser Phe Pro Pro Leu Trp Pro Ala Val
50 55 60
Ile Ala Tyr Leu Thr Trp Val Phe Phe Ile Asp Gln Ala Pro Ile His
65 70 75 80
Gly Gly Arg Ala Gln Ser Trp Leu Arg Lys Ser Arg Ile Trp Val Trp
85 90 95
Phe Ala Gly Tyr Tyr Pro Val Ser Leu Ile Lys Ser Ala Asp Leu Pro
100 105 110
Pro Asp Arg Lys Tyr Val Phe Gly Tyr His Pro His Gly Val Ile Gly
115 120 125
Met Gly Ala Ile Ala Asn Phe Ala Thr Asp Ala Thr Gly Phe Ser Thr
130 135 140
Leu Phe Pro Gly Leu Asn Pro His Leu Leu Thr Leu Gln Ser Asn Phe
145 150 155 160
Lys Leu Pro Leu Tyr Arg Glu Leu Leu Leu Ala Leu Gly Ile Cys Ser
165 170 175
Val Ser Met Lys Ser Cys Gln Asn Ile Leu Arg Gln Gly Pro Gly Ser
180 185 190
Ala Leu Thr Ile Val Val Gly Gly Ala Ala Glu Ser Leu Ser Ala His
195 200 205
Pro Gly Thr Ala Asp Leu Thr Leu Lys Arg Arg Lys Gly Phe Ile Lys
210 215 220
Leu Ala Ile Arg Gln Gly Ala Asp Leu Val Pro Val Phe Ser Phe Gly
225 230 235 240
Glu Asn Asp Ile Phe Gly Gln Leu Arg Asn Glu Arg Gly Thr Arg Leu
245 250 255
Tyr Lys Leu Gln Lys Arg Phe Gln Gly Val Phe Gly Phe Thr Leu Pro
260 265 270
Leu Phe Tyr Gly Arg Gly Leu Phe Asn Tyr Asn Val Gly Leu Met Pro
275 280 285
Tyr Arg His Pro Ile Val Ser Val Val Gly Arg Pro Ile Ser Val Glu
290 295 300
Gln Lys Asp His Pro Thr Thr Ala Asp Leu Glu Glu Val Gln Ala Arg
305 310 315 320
Tyr Ile Ala Glu Leu Lys Arg Ile Trp Glu Glu Tyr Lys Asp Ala Tyr
325 330 335
Ala Lys Ser Arg Thr Arg Glu Leu Asn Ile Ile Ala
340 345
<210> 83
<211> 2758
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<400> 83
atgcaccgct cgctcctcgt tgctgctgct gggtccaagc gaccgcgcac tgctgcgtcc 60
agtctcgctg ccgcgcgcag ggcgttccag tctgccgcgc atgtcgagaa caagctgcga 120
ctccaggtca aggaggcgga tgaggagggc gcggcgttgc acgactcgac catctcgaac 180
aaggagtacc tccagcactc gaagaacgag gtgcgtgcgg ttctagctgg ccgggacggg 240
cgggaggcgt tgttggcgaa gtcgggggcc aggagggagt gaggaggacg agggaggcga 300
cggcagctca gggaggggca gtggcaggcg gtaacggtag caagggctac taacagcagt 360
agacgaccct ctttggcccc caaacgcgcg ctgataccct tccgcccgca tcccggtcct 420
cgctcacttt cctcgacacc ctcttgatcc tcccagcgac ccatctggac cgccctgcga 480
ggtcgtgcgc tcctgaacga gccagcactg aacaaaggcg ccggtttcac gcccgaagag 540
cgggacacgt ttgggttgac gggactcttg ccgtgcgttc ttgctcgctt cgcttgaaga 600
agttgtcgag ctgaccttcc gcgaaacaga catgaggtac actcgcttga ccagcaatgc 660
aagcgcgctt attcgcagct gcaagaacgg ccgagtgctt tggcgaaata cacctgtgcg 720
ttcgcggctc ccttcttgcg tccctctcaa catggcttcg acggcgaaag ggtcgtcttt 780
gccttaacga cctcaacaag gttccgcggc ggaaccacat tcgcccatcc gctgacgccg 840
cgcccctctg gcagtcctct cctccctccg cgaccaaaac atcatcctct tctacgccct 900
ctgcctccgc cacctcaacg agctcctccc cgtcatctac actcccaccg tcggcgaagc 960
gatccaaaag tactcgacaa tctggcgcag gccggacggg ttgttcctct cctacgcaca 1020
ccggcacaag atgcgtgaga tgatgatgca ggcgaagagg cctaaggatg tcgacttgat 1080
catcgtcacc gactcggagg gcattctcgg tattggagac caaggtgtgg gagggatctt 1140
gatcgcgcag ggaaaggcga acctctacac cctcggagcc ggtatcgacc cctcgcgcat 1200
cctctcggtc gtcctcgacg tcggaaccga caactctgcg ctcttgaatg atccgctcta 1260
cctcggtttg cgacggaagc gcgtccgtgg tgccgagtac gacaagtttg tcgaccggtt 1320
ttgcgagctc gtgagggagg agtacccgca ggcgttgctg tgagtccgtt cgttcaactg 1380
cgagcggtac tgctgagcct gactttgagc tcgccgtatg cagtcacttt gaggactttg 1440
gcgtctcgaa tgcctcgaag atcctcacga cgtaccggaa caagcagtct gtctgtgcgt 1500
tcttcagcgc tcgatttgtc tccctccaaa agctcactga tccatccgca cagtcaacga 1560
cgacatgcaa ggcaccgcag cagtcgtcct cgccgccctc ctttcggccg tcaaggtcac 1620
gaagagcgag ctcaaggacc aacgcatcgt cgtctttggt ttcggcacgg ctgggtacgg 1680
gatcgcagac gggattcgga acgctttgat gcttgaggcg gggttgtcga gtgaggaggt 1740
tcggaagatc ttctggtgcg tgcgaggacc cgaagcgcga ggcaggcgag actgacgctt 1800
cctgtcgatg tgcaggtgtg tcgaccgacc gggtctgctt acgacggagc actctcccac 1860
cctacgacca ggccaagagc acttcatccg caacgcctcg gaagtctcgt cctgggagcg 1920
cgacgccgaa ggacgaatca gcttgctcga ggtcgtcaag caggccaaac cgacgattct 1980
catcggttgc tcgaccatgt cgggcgcgtt tgatgaggag gtcgtgaggg agatggcgaa 2040
gcatgtcgag cggccgatcg tcttccctct ctcaagtgcg tcgccttccc tttcctgact 2100
ccgagtaacc gctgataaag tcgatgacgt agacccgacg aagctcgccg aggctgaccc 2160
ggccgacatc aacgagtgga cgaacggact ggcgcttatg gcgaccgggt cgccgttccc 2220
gcctgtcaag acgccgcgag gcaaggagca caagatcgcc gaggcgtgag ttggcacata 2280
tccagcattc gtcgacggag cgatggctga catccgcctt ttgcagcaac aacggcctgc 2340
tttaccctgg actcggtctc gggtcagtct agctcttctc agccctcctt gctgtcgcgc 2400
tgacggtgtc tctcgcagtg tcatcgtctc tcgcgcctcg ttcttgacgg agaagatgat 2460
caccgcgggc gttgctgctc tcgccaagat ggtacgtttg cctgctcacc agcgagccat 2520
ccaaacctga ccgtctcgct cacaggctcc cgccctcgac gaccccgacg agtcgctcct 2580
ccctccactc agcgacctcc gtcatgtgtc agtcaaggtc gccacggccg tcgcgaacgc 2640
cgccaaggag gaaggcgtca gccagatcaa gcgcgacgac cccttctccc aggatgaggt 2700
tcgcgcggcg cagtgggacc ctgtctaccg cccgctcgag ctcgtcgacc gtcactag 2758
<210> 84
<211> 2232
<212> DNA
<213> Rhodotorula glutinis ATCC 204091
<220>
<221> CDS
<222> (1)..(2232)
<400> 84
atg cgc ggg agt gaa ggg ttg ccg tcg tcg tgt gtt cgt tca gcg tgt 48
Met Arg Gly Ser Glu Gly Leu Pro Ser Ser Cys Val Arg Ser Ala Cys
1 5 10 15
tcg ttt gcg ggg ttc gct cgt cgt tcc ttc gct tcc tcg tcc tca ctc 96
Ser Phe Ala Gly Phe Ala Arg Arg Ser Phe Ala Ser Ser Ser Ser Leu
20 25 30
gct cgt cgc tgt cct cct cct gct cac cct gct ccc ctc tgc cac gct 144
Ala Arg Arg Cys Pro Pro Pro Ala His Pro Ala Pro Leu Cys His Ala
35 40 45
cac ctt caa ctg aag caa gcg aag aag ccg ggt ctc tcg ctc gac tct 192
His Leu Gln Leu Lys Gln Ala Lys Lys Pro Gly Leu Ser Leu Asp Ser
50 55 60
ggc ctc gtt ccc cac cca gct cac ttc cac tct cac ttg cac tca gta 240
Gly Leu Val Pro His Pro Ala His Phe His Ser His Leu His Ser Val
65 70 75 80
gta ctc gca ccc caa cag cag cac acg gtg agc tcg cat ccc ctc cac 288
Val Leu Ala Pro Gln Gln Gln His Thr Val Ser Ser His Pro Leu His
85 90 95
tcg cgc gac cct ccc atc ccg gcg tgc gac cgc ggc acc cag tcg tgt 336
Ser Arg Asp Pro Pro Ile Pro Ala Cys Asp Arg Gly Thr Gln Ser Cys
100 105 110
gag gct gcc gca ccc tct ccc acc cct ctt tgc aca cag aca cac gat 384
Glu Ala Ala Ala Pro Ser Pro Thr Pro Leu Cys Thr Gln Thr His Asp
115 120 125
ccc gct cgc ccc atc act cgc cgg aac gct ctc ttc ctt cat ccc ccc 432
Pro Ala Arg Pro Ile Thr Arg Arg Asn Ala Leu Phe Leu His Pro Pro
130 135 140
aac tcg ctc tcc ctc tct ctc ttt cct ctc act gag cgc cca aaa gaa 480
Asn Ser Leu Ser Leu Ser Leu Phe Pro Leu Thr Glu Arg Pro Lys Glu
145 150 155 160
ctc ctc atc gag cgc gcc ttg acc cgc ctc cgc tcc atc ccg tcc gac 528
Leu Leu Ile Glu Arg Ala Leu Thr Arg Leu Arg Ser Ile Pro Ser Asp
165 170 175
ctc gag aaa tat acg ttc ctt gcg gga ttg agg tgc agg aac ccg gat 576
Leu Glu Lys Tyr Thr Phe Leu Ala Gly Leu Arg Cys Arg Asn Pro Asp
180 185 190
gtg ttt tat gga ctt gtt ggg ggg aac atg aag gag tgt tgc ggt gtt 624
Val Phe Tyr Gly Leu Val Gly Gly Asn Met Lys Glu Cys Cys Gly Val
195 200 205
ggc gag cgg tac ttg agc gag cgc ggg cgc agg agg aga agg aac ggg 672
Gly Glu Arg Tyr Leu Ser Glu Arg Gly Arg Arg Arg Arg Arg Asn Gly
210 215 220
cga gtt cga gcg gac ttg cgg ttc gcg gcg gct tct tcc ctc aac gct 720
Arg Val Arg Ala Asp Leu Arg Phe Ala Ala Ala Ser Ser Leu Asn Ala
225 230 235 240
tcc ctc gca cga aca act act cac aca ctt cga gac gac gat act gac 768
Ser Leu Ala Arg Thr Thr Thr His Thr Leu Arg Asp Asp Asp Thr Asp
245 250 255
gct ttc acc tct ccc gct cgt ctc acg aac cca cag ccg atc atc tac 816
Ala Phe Thr Ser Pro Ala Arg Leu Thr Asn Pro Gln Pro Ile Ile Tyr
260 265 270
acc ccc gtc atc ggt ctc gcg tgc caa aac tgg tct ctc atc cac cct 864
Thr Pro Val Ile Gly Leu Ala Cys Gln Asn Trp Ser Leu Ile His Pro
275 280 285
ccc ccg ccc gaa tcc gac cca aca atc gaa gcc ctc tac ctc tcc tac 912
Pro Pro Pro Glu Ser Asp Pro Thr Ile Glu Ala Leu Tyr Leu Ser Tyr
290 295 300
tcg gac ctc cct aac ctt ccc tcg ctc atc aaa ggc ctc aag acc cgt 960
Ser Asp Leu Pro Asn Leu Pro Ser Leu Ile Lys Gly Leu Lys Thr Arg
305 310 315 320
ctc ccg cac aac cag atg caa atc tcc gtc gtc act gat ggt tcg cgc 1008
Leu Pro His Asn Gln Met Gln Ile Ser Val Val Thr Asp Gly Ser Arg
325 330 335
gtt ctt gga ctt gga gat ctc gga gtt gga ggg atg ggt atc tcc caa 1056
Val Leu Gly Leu Gly Asp Leu Gly Val Gly Gly Met Gly Ile Ser Gln
340 345 350
ggg aaa ctc tcg ctt tac gtc gca gca gga ggg gtc aat cct aaa gcc 1104
Gly Lys Leu Ser Leu Tyr Val Ala Ala Gly Gly Val Asn Pro Lys Ala
355 360 365
acc ctt ccc atc gct atc gac ttc gga acc gat aac gag aag tta ctt 1152
Thr Leu Pro Ile Ala Ile Asp Phe Gly Thr Asp Asn Glu Lys Leu Leu
370 375 380
gcg gat ccg ctg tat gtc ggt cag agg atg agg agg ttg agt gag gag 1200
Ala Asp Pro Leu Tyr Val Gly Gln Arg Met Arg Arg Leu Ser Glu Glu
385 390 395 400
aaa tgc ctc gag ttt atg gac gtc ttc atg agg tgt atg cac gag acg 1248
Lys Cys Leu Glu Phe Met Asp Val Phe Met Arg Cys Met His Glu Thr
405 410 415
ttc ccg aat atg gtc atc caa cac gaa gac tgg cag acc ccg ctc gcg 1296
Phe Pro Asn Met Val Ile Gln His Glu Asp Trp Gln Thr Pro Leu Ala
420 425 430
ttc ccg ctc ttg cac aag aac cgc gac ttg tat ccg tgc ttt aat gac 1344
Phe Pro Leu Leu His Lys Asn Arg Asp Leu Tyr Pro Cys Phe Asn Asp
435 440 445
gac atc caa ggc aca ggc gcc gtc gtc cta gcc ggc gcg atc cgc gcc 1392
Asp Ile Gln Gly Thr Gly Ala Val Val Leu Ala Gly Ala Ile Arg Ala
450 455 460
ttc cac ctg aac ggc gtc gcg ctc aag gac caa aag att ctc ttc ttc 1440
Phe His Leu Asn Gly Val Ala Leu Lys Asp Gln Lys Ile Leu Phe Phe
465 470 475 480
ggc gcg ggt tcg tcg ggc gtt gga gtc gcc gag acg att tgc aag tac 1488
Gly Ala Gly Ser Ser Gly Val Gly Val Ala Glu Thr Ile Cys Lys Tyr
485 490 495
ttt gag ctg cag ggg atg agt gag cag gag gcc aag agc aag ttt tgg 1536
Phe Glu Leu Gln Gly Met Ser Glu Gln Glu Ala Lys Ser Lys Phe Trp
500 505 510
ctt gtt gac tcg aag ggc ctc gtc gcg cac aac cga ggc gac acc ctc 1584
Leu Val Asp Ser Lys Gly Leu Val Ala His Asn Arg Gly Asp Thr Leu
515 520 525
ccc tcg cac aag aaa tac ctc gcg cgc tcc gag ccc gac gcg ccg aaa 1632
Pro Ser His Lys Lys Tyr Leu Ala Arg Ser Glu Pro Asp Ala Pro Lys
530 535 540
ctc cgc tcg ctc aag gag gtc gtt gag cat gtg cag ccg act gcg ttg 1680
Leu Arg Ser Leu Lys Glu Val Val Glu His Val Gln Pro Thr Ala Leu
545 550 555 560
ttg ggg ttg tcg act gtt ggt ggg acg ttt acg aag gag atc ctc gag 1728
Leu Gly Leu Ser Thr Val Gly Gly Thr Phe Thr Lys Glu Ile Leu Glu
565 570 575
tcg atg gca acg tac aac aag cgc cct atc gtc ttc gcc ctc tcg aac 1776
Ser Met Ala Thr Tyr Asn Lys Arg Pro Ile Val Phe Ala Leu Ser Asn
580 585 590
ccc gtc gcc caa gcc gaa tgc acc ttc gaa gaa gcg atc gaa gga aca 1824
Pro Val Ala Gln Ala Glu Cys Thr Phe Glu Glu Ala Ile Glu Gly Thr
595 600 605
gac ggc cgc gtc ttg tat gcc agc ggg agc ccg ttc gat ccg gtc gag 1872
Asp Gly Arg Val Leu Tyr Ala Ser Gly Ser Pro Phe Asp Pro Val Glu
610 615 620
tac aag gag aag agg tat gag cct ggt cag ggg aac aat atg tac atc 1920
Tyr Lys Glu Lys Arg Tyr Glu Pro Gly Gln Gly Asn Asn Met Tyr Ile
625 630 635 640
ttc cct gga ctc ggg atc gga gcg atc ctc gcg cgc gtc tcc aag atc 1968
Phe Pro Gly Leu Gly Ile Gly Ala Ile Leu Ala Arg Val Ser Lys Ile
645 650 655
ccc gaa gaa ctc gtc cac gct tcg gcg cag gga ctc gcc gac tcg ctc 2016
Pro Glu Glu Leu Val His Ala Ser Ala Gln Gly Leu Ala Asp Ser Leu
660 665 670
acg ccg gag gag acg gct cgt cat ctc ctt tac ccg gac atc gag cgc 2064
Thr Pro Glu Glu Thr Ala Arg His Leu Leu Tyr Pro Asp Ile Glu Arg
675 680 685
atc cgc gaa gtc tcg atc aag atc gcc gtg acc gtc atc caa gcc gct 2112
Ile Arg Glu Val Ser Ile Lys Ile Ala Val Thr Val Ile Gln Ala Ala
690 695 700
caa aag ctc ggc gtc gac cgc aac gaa gag ctg cgc ggc aag tcg agt 2160
Gln Lys Leu Gly Val Asp Arg Asn Glu Glu Leu Arg Gly Lys Ser Ser
705 710 715 720
gcc gag att gag gcg tac gtc agg aag ggg atg tac cac ccg ctt ttg 2208
Ala Glu Ile Glu Ala Tyr Val Arg Lys Gly Met Tyr His Pro Leu Leu
725 730 735
gag gcg gag cag cag gcg cag tag 2232
Glu Ala Glu Gln Gln Ala Gln
740
<210> 85
<211> 743
<212> PRT
<213> Rhodotorula glutinis ATCC 204091
<400> 85
Met Arg Gly Ser Glu Gly Leu Pro Ser Ser Cys Val Arg Ser Ala Cys
1 5 10 15
Ser Phe Ala Gly Phe Ala Arg Arg Ser Phe Ala Ser Ser Ser Ser Leu
20 25 30
Ala Arg Arg Cys Pro Pro Pro Ala His Pro Ala Pro Leu Cys His Ala
35 40 45
His Leu Gln Leu Lys Gln Ala Lys Lys Pro Gly Leu Ser Leu Asp Ser
50 55 60
Gly Leu Val Pro His Pro Ala His Phe His Ser His Leu His Ser Val
65 70 75 80
Val Leu Ala Pro Gln Gln Gln His Thr Val Ser Ser His Pro Leu His
85 90 95
Ser Arg Asp Pro Pro Ile Pro Ala Cys Asp Arg Gly Thr Gln Ser Cys
100 105 110
Glu Ala Ala Ala Pro Ser Pro Thr Pro Leu Cys Thr Gln Thr His Asp
115 120 125
Pro Ala Arg Pro Ile Thr Arg Arg Asn Ala Leu Phe Leu His Pro Pro
130 135 140
Asn Ser Leu Ser Leu Ser Leu Phe Pro Leu Thr Glu Arg Pro Lys Glu
145 150 155 160
Leu Leu Ile Glu Arg Ala Leu Thr Arg Leu Arg Ser Ile Pro Ser Asp
165 170 175
Leu Glu Lys Tyr Thr Phe Leu Ala Gly Leu Arg Cys Arg Asn Pro Asp
180 185 190
Val Phe Tyr Gly Leu Val Gly Gly Asn Met Lys Glu Cys Cys Gly Val
195 200 205
Gly Glu Arg Tyr Leu Ser Glu Arg Gly Arg Arg Arg Arg Arg Asn Gly
210 215 220
Arg Val Arg Ala Asp Leu Arg Phe Ala Ala Ala Ser Ser Leu Asn Ala
225 230 235 240
Ser Leu Ala Arg Thr Thr Thr His Thr Leu Arg Asp Asp Asp Thr Asp
245 250 255
Ala Phe Thr Ser Pro Ala Arg Leu Thr Asn Pro Gln Pro Ile Ile Tyr
260 265 270
Thr Pro Val Ile Gly Leu Ala Cys Gln Asn Trp Ser Leu Ile His Pro
275 280 285
Pro Pro Pro Glu Ser Asp Pro Thr Ile Glu Ala Leu Tyr Leu Ser Tyr
290 295 300
Ser Asp Leu Pro Asn Leu Pro Ser Leu Ile Lys Gly Leu Lys Thr Arg
305 310 315 320
Leu Pro His Asn Gln Met Gln Ile Ser Val Val Thr Asp Gly Ser Arg
325 330 335
Val Leu Gly Leu Gly Asp Leu Gly Val Gly Gly Met Gly Ile Ser Gln
340 345 350
Gly Lys Leu Ser Leu Tyr Val Ala Ala Gly Gly Val Asn Pro Lys Ala
355 360 365
Thr Leu Pro Ile Ala Ile Asp Phe Gly Thr Asp Asn Glu Lys Leu Leu
370 375 380
Ala Asp Pro Leu Tyr Val Gly Gln Arg Met Arg Arg Leu Ser Glu Glu
385 390 395 400
Lys Cys Leu Glu Phe Met Asp Val Phe Met Arg Cys Met His Glu Thr
405 410 415
Phe Pro Asn Met Val Ile Gln His Glu Asp Trp Gln Thr Pro Leu Ala
420 425 430
Phe Pro Leu Leu His Lys Asn Arg Asp Leu Tyr Pro Cys Phe Asn Asp
435 440 445
Asp Ile Gln Gly Thr Gly Ala Val Val Leu Ala Gly Ala Ile Arg Ala
450 455 460
Phe His Leu Asn Gly Val Ala Leu Lys Asp Gln Lys Ile Leu Phe Phe
465 470 475 480
Gly Ala Gly Ser Ser Gly Val Gly Val Ala Glu Thr Ile Cys Lys Tyr
485 490 495
Phe Glu Leu Gln Gly Met Ser Glu Gln Glu Ala Lys Ser Lys Phe Trp
500 505 510
Leu Val Asp Ser Lys Gly Leu Val Ala His Asn Arg Gly Asp Thr Leu
515 520 525
Pro Ser His Lys Lys Tyr Leu Ala Arg Ser Glu Pro Asp Ala Pro Lys
530 535 540
Leu Arg Ser Leu Lys Glu Val Val Glu His Val Gln Pro Thr Ala Leu
545 550 555 560
Leu Gly Leu Ser Thr Val Gly Gly Thr Phe Thr Lys Glu Ile Leu Glu
565 570 575
Ser Met Ala Thr Tyr Asn Lys Arg Pro Ile Val Phe Ala Leu Ser Asn
580 585 590
Pro Val Ala Gln Ala Glu Cys Thr Phe Glu Glu Ala Ile Glu Gly Thr
595 600 605
Asp Gly Arg Val Leu Tyr Ala Ser Gly Ser Pro Phe Asp Pro Val Glu
610 615 620
Tyr Lys Glu Lys Arg Tyr Glu Pro Gly Gln Gly Asn Asn Met Tyr Ile
625 630 635 640
Phe Pro Gly Leu Gly Ile Gly Ala Ile Leu Ala Arg Val Ser Lys Ile
645 650 655
Pro Glu Glu Leu Val His Ala Ser Ala Gln Gly Leu Ala Asp Ser Leu
660 665 670
Thr Pro Glu Glu Thr Ala Arg His Leu Leu Tyr Pro Asp Ile Glu Arg
675 680 685
Ile Arg Glu Val Ser Ile Lys Ile Ala Val Thr Val Ile Gln Ala Ala
690 695 700
Gln Lys Leu Gly Val Asp Arg Asn Glu Glu Leu Arg Gly Lys Ser Ser
705 710 715 720
Ala Glu Ile Glu Ala Tyr Val Arg Lys Gly Met Tyr His Pro Leu Leu
725 730 735
Glu Ala Glu Gln Gln Ala Gln
740
<210> 86
<211> 3875
<212> DNA
<213> Rhodosporidium toruloides
<400> 86
gggccgaggg caagcagtgg atcgctgagc gcgcgggcaa gcaggttcag gtgcgtttcc 60
ctcttccgcc tcgcctcctt tttttgcaga aagaacacgc gcactgacac tgcgaacgaa 120
cgcgcacgcg caggtcgaga agacgacagg cacgctcaac aacttcatcg tcgagccgtt 180
ctgcccgcac ccttcggacg ccgagtacta catctgcatc aactcggtcc gcgagggcga 240
cgtgatcctg ttcacgcacg agggcggtgt cgacgtcggc gatgtcgacg ccaaggcgct 300
cacgctcctc gtccccgtcg gcggcgagct cccctcgcgc gacgagatcc gctcccagct 360
cctcaagcac gtcactggcg ccgagcgcca ggaggccctc atcgactaca tcatccgcct 420
ctactcggtc tacgtcgacc tccactttgc ctacctcgag atcaaccctc tcgtcgccgt 480
cgagaacccc tcgactggca agaccgacat cttctacctc gacatggccg ccaagctcga 540
ccagacggcc gagtacgtcg tcggtcccaa gtgggccatc gcgcgcgacc cgtccatcat 600
caaccccgcc gctgcaccca tgtcgaacgg caagatttcg gccgacaagg gcccgcccat 660
gttctggccc cctcccttcg gtcgtgactt gaccaaggag gaggcctaca tcgccaagct 720
cgacggctcg accggcgcat ccctcaagct caccgtcttg aacgccgagg gacgcatctg 780
gacgatggtt gccggtggtg gtgcttcggt cgtctactcg gacgccatcg ccgcgcacgg 840
attcgcgcac gagctcgcca actacggcga gtactcgggc gcgcccaccc agacccagac 900
ttacgagtac gccaagacca tccgtgcgtt tccgccttct ctcttccgct ctcagacagc 960
ctcgctgacg ctccgtaacg ctcgcagtcg acctgatgac ccgcggcacg cccaacccgc 1020
agggcaagct cctcttcatc ggcggcggta tcgccaactt caccaacgtc gccgcgacct 1080
tcaagggcat catcacggcg ctcaaggagt accagcaccg tctccaggag cacaaggtcc 1140
gcatctttgt ccgccgcggc ggccccaact accaggaggg cctcaaggcc atgcgcctcc 1200
tcggcgagac gctcggcgtc gagatccagg tctttggccc cgaaacccac atcacctcga 1260
tcgtcccgct cggcctcggc ttgatcaagt cggtcgacga cgccctcaag gttcccggcg 1320
cccgctccgc cgccgacgcg accggcaccc tcacccccgt tcccggctcg cccaagtcgc 1380
gcgccgccca gctcccgacc ggcgcgtcga cgccctcgcg cccgcagccc caggacaaca 1440
tcgtcagctt ctcggacaag atccacgcgc ccgactcggg ccgcccgtgg taccgcccct 1500
ttgacgagac gacgcgctcg atcgtctacg gtctccagcc ccgcgcgatc cagggcatgc 1560
tcgactttga ctttgcctgc ggccgcgaga cgccctcggt cgccgccatg gtctacccct 1620
tcggcggcca ccacgtccag aagttctact ggggcaccaa ggagaccctc ctccccgtct 1680
tcacctcgat gaaggaggcc gtcgccaagt gccccgacgc cgacgtcgtc gtcaactttg 1740
cgtcatcgcg ctcggtctac cagtcgaccc tcgaggcgct cgagttcccc cagatcaagg 1800
ccatcgccct catcgccgag ggtgtccccg agcgccacgc ccgcgagatc ctccacctcg 1860
ccaagaagaa ggaggtcatc atcatcggtc ccgcgacggt cggcggcatc aagcccggct 1920
gcttccgtat cggcaacacg ggcggcatga acgagaacat cctctcgtcc aagctttacc 1980
gcgccggttc cgtcggctac gtctccaagt ccggaggcat gtcgaacgag ctcaacaaca 2040
tcctttcgct cacgaccgac ggcgcgtacg agggcatcgc catcggcggt gaccgctacc 2100
cgggcaccac cttcatcgac caccttctcc gctacgaggc cgacccgaac tgcaagatgc 2160
tcgtcctcct cggagaggtc ggcggtgtcg aggagtaccg cgtcatcgag gccgtcaagt 2220
cgggccagat caagaagccc atcgtcgcgt gggccatcgg cacctgcgcc aagatgtttg 2280
cgaccgacgt ccagttcggc cacgccggtt ccatggccaa ctcggacctc gagaccgccg 2340
aggccaagaa caacgccatg cgcgccgccg gcttcatcgt ccccccgacc ttcgaggagc 2400
tcccgcaggt cctcgccgag acctaccaga agctcgtcgg cgacggcacg atccagccca 2460
agcccgaggt tcctccccct cagatcccga tggactacaa ctgggctcag acgctcggca 2520
tggtccgcaa gcccgccgcc ttcatctcga ccatctcgga cgagcgcggc caggagctcc 2580
tctacgccgg catgcccatc tccaaggtct tcgaggagga catcggcatc ggcggcgtcg 2640
tctcgctcct ctggttcaag cgccgcctcc ccgcctacgc gaccaagttc ctcgagatgg 2700
tcctcatgct cacggccgac cacggtcccg ccgtctcggg cgccatgacc accgtcatca 2760
ccacccgtgc cggcaaggac ctcgtctcgt cgctcgtcgc cggtctcctc accatcggcg 2820
accgcttcgg tggcgcgctc gacggcgccg cgcaggagtt cacgcgcgct ttcgaggccg 2880
gcctcacgcc ccgcgagttt gtcgactcga tgcgcaaggc caacaagctc atccccggta 2940
tcggccacaa ggtcaagtcc aaggccaacc cggacaagcg cgtcgagctc gtcaagaact 3000
acgtcttcaa gcacttcccg tcgacaaagc tcctcgagta cgctctcgct gtcgaggacg 3060
tcacgagcgc gaagaaggac accctcatcc tcaacgtcga cggaggtgag ccttcgcttg 3120
ctcctcctcg acttatcctt gtactgacgc ttcctcctcg tccagcaatc gccgtggcct 3180
tctgcgattt gctcaaaaag tagcgctctt tcactgcgtc aaaaacagga cgaaggagaa 3240
ctgacccgcc ccgcttcctt cccgcagctc gggcgcgttc accaccgagg aggctgccga 3300
gtacatgaag gtgtgtcctt gctcttctat agcgttcgtg cgaccgaacg aaggagcgtg 3360
gaacgttcag aggcgcggga aggaagagga ggggagacgg ggcgaacgga attgctgacc 3420
tcgcgccttc gcagatcgga accttgaacg gtctcttcgt tcttggtcgc tcgatcggct 3480
tcattgccca ccacctcgac cagaagcgcc tcaagcagcc gctctaccgt cacccggccg 3540
agtgcgctcc atctcctcct tgccatctcg acctttccgc tgacctgctc ctcctccgca 3600
gcgacatctt catccagccc ttcaacaccg accgcatcct cgtccagcag cgccagtaaa 3660
tggcgcacgc tcgatgcgtc accgccgtgg ccgcctagac ttcgctcgca tccgcctctc 3720
tcgttctcgc ctcgttcgac attagagttc ccttcttgtt ttcgcgtcgt tccgtctttc 3780
ttgctttctt gttctttctc tttttctcgc tctctttccg gcagtacctc ctctcatggg 3840
ctctcagtcg tcgtgcatca aagtttcgtt tctcc 3875
<210> 87
<211> 3489
<212> DNA
<213> Rhodosporidium toruloides
<400> 87
gggccgaggg caagcagtgg atcgctgagc gcgcgggcaa gcaggttcag gtgcgtttcc 60
ctcttccgcc tcgcctcctt tttttgcaga aagaacacgc gcactgacac tgcgaacgaa 120
cgcgcacgcg caggtcgaga agacgacagg cacgctcaac aacttcatcg tcgagccgtt 180
ctgcccgcac ccttcggacg ccgagtacta catctgcatc aactcggtcc gcgagggcga 240
cgtgatcctg ttcacgcacg agggcggtgt cgacgtcggc gatgtcgacg ccaaggcgct 300
cacgctcctc gtccccgtcg gcggcgagct cccctcgcgc gacgagatcc gctcccagct 360
cctcaagcac gtcactggcg ccgagcgcca ggaggccctc atcgactaca tcatccgcct 420
ctactcggtc tacgtcgacc tccactttgc ctacctcgag atcaaccctc tcgtcgccgt 480
cgagaacccc tcgactggca agaccgacat cttctacctc gacatggccg ccaagctcga 540
ccagacggcc gagtacgtcg tcggtcccaa gtgggccatc gcgcgcgacc cgtccatcat 600
caaccccgcc gctgcaccca tgtcgaacgg caagatttcg gccgacaagg gcccgcccat 660
gttctggccc cctcccttcg gtcgtgactt gaccaaggag gaggcctaca tcgccaagct 720
cgacggctcg accggcgcat ccctcaagct caccgtcttg aacgccgagg gacgcatctg 780
gacgatggtt gccggtggtg gtgcttcggt cgtctactcg gacgccatcg ccgcgcacgg 840
attcgcgcac gagctcgcca actacggcga gtactcgggc gcgcccaccc agacccagac 900
ttacgagtac gccaagacca tcctcgacct gatgacccgc ggcacgccca acccgcaggg 960
caagctcctc ttcatcggcg gcggtatcgc caacttcacc aacgtcgccg cgaccttcaa 1020
gggcatcatc acggcgctca aggagtacca gcaccgtctc caggagcaca aggtccgcat 1080
ctttgtccgc cgcggcggcc ccaactacca ggagggcctc aaggccatgc gcctcctcgg 1140
cgagacgctc ggcgtcgaga tccaggtctt tggccccgaa acccacatca cctcgatcgt 1200
cccgctcggc ctcggcttga tcaagtcggt cgacgacgcc ctcaaggttc ccggcgcccg 1260
ctccgccgcc gacgcgaccg gcaccctcac ccccgttccc ggctcgccca agtcgcgcgc 1320
cgcccagctc ccgaccggcg cgtcgacgcc ctcgcgcccg cagccccagg acaacatcgt 1380
cagcttctcg gacaagatcc acgcgcccga ctcgggccgc ccgtggtacc gcccctttga 1440
cgagacgacg cgctcgatcg tctacggtct ccagccccgc gcgatccagg gcatgctcga 1500
ctttgacttt gcctgcggcc gcgagacgcc ctcggtcgcc gccatggtct accccttcgg 1560
cggccaccac gtccagaagt tctactgggg caccaaggag accctcctcc ccgtcttcac 1620
ctcgatgaag gaggccgtcg ccaagtgccc cgacgccgac gtcgtcgtca actttgcgtc 1680
atcgcgctcg gtctaccagt cgaccctcga ggcgctcgag ttcccccaga tcaaggccat 1740
cgccctcatc gccgagggtg tccccgagcg ccacgcccgc gagatcctcc acctcgccaa 1800
gaagaaggag gtcatcatca tcggtcccgc gacggtcggc ggcatcaagc ccggctgctt 1860
ccgtatcggc aacacgggcg gcatgaacga gaacatcctc tcgtccaagc tttaccgcgc 1920
cggttccgtc ggctacgtct ccaagtccgg aggcatgtcg aacgagctca acaacatcct 1980
ttcgctcacg accgacggcg cgtacgaggg catcgccatc ggcggtgacc gctacccggg 2040
caccaccttc atcgaccacc ttctccgcta cgaggccgac ccgaactgca agatgctcgt 2100
cctcctcgga gaggtcggcg gtgtcgagga gtaccgcgtc atcgaggccg tcaagtcggg 2160
ccagatcaag aagcccatcg tcgcgtgggc catcggcacc tgcgccaaga tgtttgcgac 2220
cgacgtccag ttcggccacg ccggttccat ggccaactcg gacctcgaga ccgccgaggc 2280
caagaacaac gccatgcgcg ccgccggctt catcgtcccc ccgaccttcg aggagctccc 2340
gcaggtcctc gccgagacct accagaagct cgtcggcgac ggcacgatcc agcccaagcc 2400
cgaggttcct ccccctcaga tcccgatgga ctacaactgg gctcagacgc tcggcatggt 2460
ccgcaagccc gccgccttca tctcgaccat ctcggacgag cgcggccagg agctcctcta 2520
cgccggcatg cccatctcca aggtcttcga ggaggacatc ggcatcggcg gcgtcgtctc 2580
gctcctctgg ttcaagcgcc gcctccccgc ctacgcgacc aagttcctcg agatggtcct 2640
catgctcacg gccgaccacg gtcccgccgt ctcgggcgcc atgaccaccg tcatcaccac 2700
ccgtgccggc aaggacctcg tctcgtcgct cgtcgccggt ctcctcacca tcggcgaccg 2760
cttcggtggc gcgctcgacg gcgccgcgca ggagttcacg cgcgctttcg aggccggcct 2820
cacgccccgc gagtttgtcg actcgatgcg caaggccaac aagctcatcc ccggtatcgg 2880
ccacaaggtc aagtccaagg ccaacccgga caagcgcgtc gagctcgtca agaactacgt 2940
cttcaagcac ttcccgtcga caaagctcct cgagtacgct ctcgctgtcg aggacgtcac 3000
gagcgcgaag aaggacaccc tcatcctcaa ccaatcgccg tggccttctg cgatttgctc 3060
aaaaactcgg gcgcgttcac caccgaggag gctgccgagt acatgaagat cggaaccttg 3120
aacggtctct tcgttcttgg tcgctcgatc ggcttcattg cccaccacct cgaccagaag 3180
cgcctcaagc agccgctcta ccgtcacccg gccgacgaca tcttcatcca gcccttcaac 3240
accgaccgca tcctcgtcca gcagcgccag taaatggcgc acgctcgatg cgtcaccgcc 3300
gtggccgcct agacttcgct cgcatccgcc tctctcgttc tcgcctcgtt cgacattaga 3360
gttcccttct tgttttcgcg tcgttccgtc tttcttgctt tcttgttctt tctctttttc 3420
tcgctctctt tccggcagta cctcctctca tgggctctca gtcgtcgtgc atcaaagttt 3480
cgtttctcc 3489
<210> 88
<211> 1148
<212> PRT
<213> Rhodosporidium toruloides
<400> 88
Met Ser Ala Lys Pro Ile Arg Glu Tyr Asp Ala Lys Leu Leu Leu Ala
1 5 10 15
Tyr His Leu Ala Arg Ala Pro Thr Ala Gly Ser Lys Ala Val Ala Arg
20 25 30
Asp Gly Phe Gln Ser Pro Glu Val Lys Val Ala Gln Val Ser Trp Asp
35 40 45
Pro Glu Thr Asn Gln Val Thr Pro Asp Ala Ala Leu Pro His Trp Val
50 55 60
Phe Thr Glu Lys Leu Val Val Lys Pro Asp Gln Leu Ile Lys Arg Arg
65 70 75 80
Gly Lys Ala Gly Leu Leu Ala Leu Asn Lys Thr Trp Ala Glu Gly Lys
85 90 95
Gln Trp Ile Ala Glu Arg Ala Gly Lys Gln Val Gln Val Glu Lys Thr
100 105 110
Thr Gly Thr Leu Asn Asn Phe Ile Val Glu Pro Phe Cys Pro His Pro
115 120 125
Ser Asp Ala Glu Tyr Tyr Ile Cys Ile Asn Ser Val Arg Glu Gly Asp
130 135 140
Val Ile Leu Phe Thr His Glu Gly Gly Val Asp Val Gly Asp Val Asp
145 150 155 160
Ala Lys Ala Leu Thr Leu Leu Val Pro Val Gly Gly Glu Leu Pro Ser
165 170 175
Arg Asp Glu Ile Arg Ser Gln Leu Leu Lys His Val Thr Gly Ala Glu
180 185 190
Arg Gln Glu Ala Leu Ile Asp Tyr Ile Ile Arg Leu Tyr Ser Val Tyr
195 200 205
Val Asp Leu His Phe Ala Tyr Leu Glu Ile Asn Pro Leu Val Ala Val
210 215 220
Glu Asn Pro Ser Thr Gly Lys Thr Asp Ile Phe Tyr Leu Asp Met Ala
225 230 235 240
Ala Lys Leu Asp Gln Thr Ala Glu Tyr Val Val Gly Pro Lys Trp Ala
245 250 255
Ile Ala Arg Asp Pro Ser Ile Ile Asn Pro Ala Ala Ala Pro Met Ser
260 265 270
Asn Gly Lys Ile Ser Ala Asp Lys Gly Pro Pro Met Phe Trp Pro Pro
275 280 285
Pro Phe Gly Arg Asp Leu Thr Lys Glu Glu Ala Tyr Ile Ala Lys Leu
290 295 300
Asp Gly Ser Thr Gly Ala Ser Leu Lys Leu Thr Val Leu Asn Ala Glu
305 310 315 320
Gly Arg Ile Trp Thr Met Val Ala Gly Gly Gly Ala Ser Val Val Tyr
325 330 335
Ser Asp Ala Ile Ala Ala His Gly Phe Ala His Glu Leu Ala Asn Tyr
340 345 350
Gly Glu Tyr Ser Gly Ala Pro Thr Gln Thr Gln Thr Tyr Glu Tyr Ala
355 360 365
Lys Thr Ile Leu Asp Leu Met Thr Arg Gly Thr Pro Asn Pro Gln Gly
370 375 380
Lys Leu Leu Phe Ile Gly Gly Gly Ile Ala Asn Phe Thr Asn Val Ala
385 390 395 400
Ala Thr Phe Lys Gly Ile Ile Thr Ala Leu Lys Glu Tyr Gln His Arg
405 410 415
Leu Gln Glu His Lys Val Arg Ile Phe Val Arg Arg Gly Gly Pro Asn
420 425 430
Tyr Gln Glu Gly Leu Lys Ala Met Arg Leu Leu Gly Glu Thr Leu Gly
435 440 445
Val Glu Ile Gln Val Phe Gly Pro Glu Thr His Ile Thr Ser Ile Val
450 455 460
Pro Leu Gly Leu Gly Leu Ile Lys Ser Val Asp Asp Ala Leu Lys Val
465 470 475 480
Pro Gly Ala Arg Ala Ala Ala Asp Ala Thr Gly Thr Leu Thr Pro Val
485 490 495
Pro Gly Ser Pro Lys Ser Arg Ala Ala Gln Leu Pro Thr Gly Ala Ser
500 505 510
Thr Pro Ser Arg Gln Gln Pro Gln Asp Asn Ile Val Ser Phe Ser Asp
515 520 525
Lys Val His Ala Pro Asp Ser Gly Arg Pro Trp Tyr Arg Pro Phe Asp
530 535 540
Glu Thr Thr Arg Ser Ile Val Tyr Gly Leu Gln Pro Arg Ala Ile Gln
545 550 555 560
Gly Met Leu Asp Phe Asp Phe Ala Cys Gly Arg Glu Thr Pro Ser Val
565 570 575
Ala Ala Met Val Tyr Pro Phe Gly Gly His His Val Gln Lys Phe Tyr
580 585 590
Trp Gly Thr Lys Glu Thr Leu Leu Pro Val Phe Thr Ser Met Lys Glu
595 600 605
Ala Val Ala Lys Cys Pro Asp Ala Asp Val Val Val Asn Phe Ala Ser
610 615 620
Ser Arg Ser Val Tyr Gln Ser Thr Leu Glu Ala Leu Glu Phe Pro Gln
625 630 635 640
Ile Lys Ala Ile Ala Leu Ile Ala Glu Gly Val Pro Glu Arg His Ala
645 650 655
Arg Glu Ile Leu His Leu Ala Lys Lys Lys Glu Val Ile Ile Ile Gly
660 665 670
Pro Ala Thr Val Gly Gly Ile Lys Pro Gly Cys Phe Arg Ile Gly Asn
675 680 685
Thr Gly Gly Met Asn Glu Asn Ile Leu Ser Ser Lys Leu Tyr Arg Ala
690 695 700
Gly Ser Val Gly Tyr Val Ser Lys Ser Gly Gly Met Ser Asn Glu Leu
705 710 715 720
Asn Asn Ile Leu Ser Leu Thr Thr Asp Gly Ala Tyr Glu Gly Ile Ala
725 730 735
Ile Gly Gly Asp Arg Tyr Pro Gly Thr Thr Phe Ile Asp His Leu Leu
740 745 750
Arg Tyr Glu Ala Asp Pro Asn Cys Lys Met Leu Val Leu Leu Gly Glu
755 760 765
Val Gly Gly Val Glu Glu Tyr Arg Val Ile Glu Ala Val Lys Ser Gly
770 775 780
Gln Ile Lys Lys Pro Ile Val Ala Trp Ala Ile Gly Thr Cys Ala Lys
785 790 795 800
Met Phe Ala Thr Asp Val Gln Phe Gly His Ala Gly Ser Met Ala Asn
805 810 815
Ser Asp Leu Glu Thr Ala Glu Ala Lys Asn Asn Ala Met Arg Ala Ala
820 825 830
Gly Phe Ile Val Pro Pro Thr Phe Glu Glu Leu Pro Gln Val Leu Ala
835 840 845
Glu Thr Tyr Gln Lys Leu Val Gly Asp Gly Thr Ile Gln Pro Lys Pro
850 855 860
Glu Val Pro Pro Pro Gln Ile Pro Met Asp Tyr Asn Trp Ala Gln Thr
865 870 875 880
Leu Gly Met Val Arg Lys Pro Ala Ala Phe Ile Ser Thr Ile Ser Asp
885 890 895
Glu Arg Gly Gln Glu Leu Leu Tyr Ala Gly Met Pro Ile Ser Lys Val
900 905 910
Phe Glu Glu Asp Ile Gly Ile Gly Gly Val Val Ser Leu Leu Trp Phe
915 920 925
Lys Arg Arg Leu Pro Ala Tyr Ala Thr Lys Phe Leu Glu Met Val Leu
930 935 940
Met Leu Thr Ala Asp His Gly Pro Ala Val Ser Gly Ala Met Thr Thr
945 950 955 960
Val Ile Thr Thr Arg Ala Gly Lys Asp Leu Val Ser Ser Leu Val Ala
965 970 975
Gly Leu Leu Thr Ile Gly Asp Arg Phe Gly Gly Ala Leu Asp Gly Ala
980 985 990
Ala Gln Glu Phe Thr Arg Ala Phe Glu Ala Gly Leu Thr Pro Arg Glu
995 1000 1005
Phe Val Asp Ser Met Arg Lys Ala Asn Lys Leu Ile Pro Gly Ile
1010 1015 1020
Gly His Lys Val Lys Ser Lys Ala Asn Pro Asp Lys Arg Val Glu
1025 1030 1035
Leu Val Lys Asn Tyr Val Phe Lys His Phe Pro Ser Ala Lys Leu
1040 1045 1050
Leu Glu Tyr Ala Leu Ala Val Glu Asp Val Thr Ser Ala Lys Lys
1055 1060 1065
Asp Thr Leu Ile Leu Asn Gln Ser Arg Phe Leu Ser Ser Gly Ala
1070 1075 1080
Phe Thr Ala Glu Glu Ala Ala Glu Tyr Met Lys Ile Gly Thr Leu
1085 1090 1095
Asn Gly Leu Phe Val Leu Gly Arg Ser Ile Gly Phe Ile Ala His
1100 1105 1110
His Leu Asp Gln Lys Arg Leu Lys Gln Pro Leu Tyr Arg His Pro
1115 1120 1125
Ala Asp Asp Ile Phe Ile Gln Pro Phe Asn Thr Asp Arg Ile Leu
1130 1135 1140
Val Gln Gln Arg Gln
1145
<210> 89
<211> 7222
<212> DNA
<213> Rhodosporidium toruloides
<400> 89
accacactct cccgcttgcg ggctctcttt ctcgcttggc gctcctgcta ccgctactct 60
agactctcct agtctccctg cacaaccatc cctatcccct ccgcctctct cgcacacccc 120
ccacagcttc gttccccaac ttcacttccg atgccgtgcg tcgcctccct ttcgcctggc 180
gggcccgcgc ctgcttccga ggacaactac tgattgtggg atcatgcgac gacaggttct 240
ctggcgaggc gaaggcggtc aacggatcgc actcggtcga cgaggcgccg aagaacccca 300
agtacgacca tgggcgggtc gtaaagtacc tcggtgcgtt tcatggggtc ctttgagcac 360
tctggagact ttctggaccg gcggacgggc agttgggctg accttgcatt tcctctcctc 420
ctcgacttcg ctcacctcga caggcggcaa ctcgctcgag tctgcgcccc cttccaaggt 480
cgctgactgg gtcagggagc gtggtggaca caccgtcatc acaaaggtgc gctcactcgg 540
acccctcttg caccgcccct cccgaccgtc cccaactcgc tcgtcctgac ccaactcgcg 600
ctccgcacag atcctgatcg ccaacaatgg tatcgctgca gtcaaggaga tccgctcggt 660
gcgcaagtgg gcgtacgaga cgttcggaag cgagcgcgcg atcgagttta ccgtcatggc 720
gaccccggag gacctcaagg tcaatgcaga ctacatccgc atggccgacc agtacgtcga 780
ggtacccggt ggaaccaaca acaacaacta cgcgaacgtc gatgtcatcg ttgatgttgc 840
cgagcgcgct ggcgtccacg ccgtctgggc aggatggttc gtcccgtttc cttcccgctc 900
ccttcagcgg aagtgcccgt gctaacttgc cttcgacgga cacaggggcc acgcttccga 960
gaacccccgc cttcccgagt cgctcgccgc ctcgaagcac aagatcgtct tcatcggtcc 1020
tcccggctcc gccatgcgct cgctcggcga caagatctcg tcgaccatcg ttgcgcagca 1080
cgcccaggtt ccgtgcatgg actggtccgg acagggcgtc gaccaggtca cccagtcgcc 1140
cgagggctac gttaccgtcg ccgacgacgt ctaccagcag gcctgtgtgc acgacgccga 1200
cgagggtctc gcccgcgcgt cgaggatcgg ataccccgtc atgatcaagg cgtccgaggg 1260
aggaggagga aagggtattc gcaaggtcga gaaggagcag gacttcaagc aggctttcca 1320
ggccgtcctc accgaggttc ccggtgcgtc gattcgcttt cagtttgaag catccttagc 1380
tgactcgccg cttccatagg ctcgcccgtc ttcatcatga agctcgctgg cgctgctcgc 1440
cacctcgagg ttcaggttct cgccgaccag tacggcaacg ccatctcgct cttcggccgt 1500
gactgctcgg tccagcgtcg ccaccagaag atcatcgaag aggcgcccgt caccatcgcc 1560
aagcccgaca cgttcgagca gatggaaaag tcggccgtcc gccttgccaa gctcgtcggt 1620
tacgtctcgg cgggaaccgt cgagttcctc tactcggctg ccgacgacaa gttcgccttc 1680
ctcgagctca acccgcgtct ccaggttgag cacccgacga ccgagatggt ctcgggcgtc 1740
aaccttcccg ccgctcagct ccaggtcgcc atgggtgtcc ctctccaccg catccgcgac 1800
atccgcaccc tctacggcaa ggcacccaac ggcagcagcg agatcgactt cgacttcgag 1860
aatcccgaat cggccaagac gcagcgcaag ccctcgccga agggtcacgt cgtcgccgtc 1920
cgtatcacgg ctgagaaccc tgatgccggc ttcaagccgt cgatgggtac cctccaggag 1980
ctcaacttcc gttcgagcac gaacgtctgg ggttacttct ccgtcggcag cgccggtggt 2040
ctccacgagt ttgccgactc gcagtttggc cacatctttg cgtacggctc ggaccgttcc 2100
gagtcgcgca agaacatggt cgtcgcgctc aaggagctct cgattcgcgg tgacttccgc 2160
acgaccgtcg agtacctcat caagcttctc gagaccgatg cgttcgagca gaacacgatt 2220
acgaccgcct ggctcgacag cctcatctcg gctcgcctga ccgccgagag gcccgacacg 2280
actctcgcca tcatctgcgg cgccgtcacg aaggctcacc tcgcttccga ggccaacatc 2340
gccgagtaca agcgcatcct cgagaagggt cagagccccg ccaaggagct cctggccacc 2400
gtcgtccctc tcgagttcgt cctcgaggac gtcaagtacc gcgcgaccgc ctcgcgctcg 2460
tcgccttcga gctggtccat ctacgtcaac ggctcgaacg tctcggtcgg catccgccct 2520
ctcgccgatg gcggtctcct catcctcctc gacggccgct catacacctg ctacgccaag 2580
gaggaggtcg gcgcgcttcg cctctcaatc gactcgagga ctgtcctcat cgctcaggag 2640
aacgacccca cccagcttcg ctcgccatcg cccggcaagc tcgtccgcta cttcatcgag 2700
tcgggcgagc acatctcaaa gggcgaggcg tacgccgaga tcgaggtcat gaaggtgcgt 2760
cgccccggca ttccctgctc gtgttcgtct cgctgacgct cgatgcctcg cagatgatca 2820
tgcccctcat cgctgctgag gacggtatcg cgcagttcat caagcagccg ggagcgacgc 2880
tcgaggctgg cgacatcctg ggtatcttgt cgctcgacga cccgagccgc gtccaccacg 2940
ccaagccgtt cgacggccag cttccggccc ttggcttgcc ttccatcatc ggcaacaagc 3000
cgcaccagcg cttcgcctac ctcaaggacg tcctctcgaa catcctcatg ggctacgaca 3060
accaggctgt gatgcagtcg agcatcaagg agctcatctc ggtccttcgc aaccccgagc 3120
tcccgtacgg cgaggccaac gccgtcctct caacgctttc gggccgtatc cccgccaagc 3180
tcgagcagac cctccgccag tacattgacc aagctcacga gtctggcgcc gagttcccgt 3240
ccgccaagtg ccgcaaggcg atcgacacga ccctcgagca gctccgccct gccgaggcgc 3300
agaccgtccg caacttcctc gtcgcgttcg acgacatcgt ctaccgctac cgctcgggcc 3360
tcaagcacca cgagtggtcg acgctcgccg gcatctttgc cgcgtacgcc gagacggaga 3420
agccgttcag cggcaaggac ggcgacgtcg tcctcgagct ccgcgacgcc caccgcgact 3480
cgctcgactc ggtcgtcaag atcgtcctct cgcactacaa ggccgcctcg aagaactcgc 3540
tcgtcctcgc cctcctcgac attgtcaagg actcggactc ggtcccgctc atcgagcagg 3600
tcgtcagccc cgcgctcaag gaccttgccg acctcgattc gaaggccacg accaaggtcg 3660
ccctcaaggc tcgcgaggtg ctcatccaca tccagctccc ctcgctcgac gagcgcctcg 3720
gccagcttga gcagatcctc aaggcctcgg tcacgcccac cgtctacggc gagcctggcc 3780
acgatcgcac tcctcgcggg gaggtcctca aggatgtcat cgactcgcgc ttcaccgtct 3840
tcgacgttct cccgagcttc ttccagcacc aggaccactg ggtctcgctt gccgcgctcg 3900
acacctacgt ccgccgcgct taccgctcgt acaatctcct caacatcgag cacattgagg 3960
ccgatgctgc cgaggacgag cccgcgaccg tcgcctggtc gttccgcatg cgtaaggctg 4020
cgtccgaatc cgagccgcct acgcccacga ccggcctcac ctcgcagcgc accgcctcgt 4080
actcggactt gaccttcctc ctcaacaacg cccagtccga gccgatccgc tacggcgcga 4140
tgttctcggt ccgctcgctc gaccgcttcc gccaggagct cggcaccgtc ctccgccact 4200
tccccgactc gaacaagggc aagctccagc agcagcctgc cgcttcgtcg agccaggagc 4260
agtggaacgt catcaatgtt gcgctcacgg tccccgccag cgcgcaggtc gacgaggacg 4320
ctctccgtgc cgacttcgcc gctcacgtca acgcgatgag cgccgagatc gacgctcgcg 4380
gcatgcgccg cctcaccctc ctcatctgcc gcgagggcca gtacccgtcc tactacaccg 4440
tccgcaagca ggacggcacc tggaaggagc tcgagacgat ccgcgacatc gagcctgccc 4500
tcgccttcca gctcgagctc ggccgcctct ccaacttcca cctcgagccg tgcccagtcg 4560
agaaccgcca ggttcacgtc tactacgcga ccgccaaggg caactcgtcc gactgccgct 4620
tcttcgtccg tgcgctcgtc cgccctggcc gcctccgcgg caacatgaag acggccgact 4680
accttgtgtc cgaggccgac cgcctcgtca ccgacgtcct cgactcgctc gaggtcgcca 4740
gctcgcagcg ccgcgccgcc gacggcaacc acatctcggt gcgtactcgc agccttgccc 4800
tccctcgttc tcgcgctgac tcgctctcgt ttcgcagctc aacttcctgt actcgctccg 4860
tctcgacttt gacgaggttc aggctgccct cgccggcttc atcgaccgcc acggcaagcg 4920
cttctggcgt ctccgcgtca ccggcgccga gatccgcatc gtcctcgagg acgcgcaggg 4980
caacatccag cccatccgcg ccatcattga gaacgtctcg ggcttcgtcg tcaagtacga 5040
ggcgtaccgc gaggtcacga ctgacaaggg ccaggtcatc ctcaagtcga tcggtccgca 5100
gggcgccctg caccttcagc cggtcaactt cccctacccg accaaggagt ggcttcagcc 5160
gaagcgctac aaggcccacg tcgtcggcac gacgtacgtc tacgacttcc ccgacctttt 5220
ccgccaggct atccgcaagc agtggaaggc cgccggcaag accgcgcccg ccgagctcct 5280
cgtcgccaag gagctcgtcc tcgacgaatt cggcaagccc caggaggtcg cccgcccgcc 5340
tggcactaac aacatcggca tggtcggctg gatctacaca atcttcacgc ccgagtaccc 5400
gaccggccgc cgcgtcgtcg tcatcgccaa cgacatcacc ttcaagattg gttcgttcgg 5460
ccccgaggag gaccgctact tcttcgccgt cacgcagctc gcgcgccagc ttggcttgcc 5520
gcgcgtctac ctctcggcca actcgggcgc tcgcctcggt attgccgagg agctcgtcga 5580
cttgttcagc gtcgcgtggg tcgacagctc gcggccggag aagggcttca agtacctcta 5640
cctcaccgcc gagaagctcg gcgagctcaa gaacaagggc gagaagagcg tcattacgaa 5700
gcgcatcgag gacgagggcg agacgcgcta ccagatcacc gacatcatcg gcttgcagga 5760
gggtctcggt gtcgagtcgc tcaagggctc tggcctcatc gccggtgaga cctcgcgcgc 5820
gtacgacgac atcttcacga tcacgctcgt caccgcccgc tcggtcggta tcggtgcgta 5880
cctcgtccgc ctcggccagc gtgccgtcca ggtcgagggc cagccgatca tcctcaccgg 5940
tgccggcgcg ctcaacaagg tcctcggtcg cgaggtctac tcgtccaact tgcagctcgg 6000
cggcacgcaa atcatgtaca agaacggtgt ctcgcacttg acggccgcca acgacctcga 6060
gggtgtcctc agcatcgtcc agtggctcgc ctttgtcccc gagcaccgcg gcgcgcctct 6120
cccgatcatg ccttcgcccg tcgacccgtg ggaccgctcg atcgactaca cgcccatcaa 6180
gggcgcgtac gacccgcgct ggttcctcgc cggcaagacg gacgaggccg acggccgctg 6240
gctctctggc ttcttcgaca agggctcgtt ccaggagacg ctctcgggct gggcgcagac 6300
cgtcgtcgtc ggccgcgctc gcctcggcgg catccccatg ggcgctatcg cagtcgaaac 6360
ccgcaccatc gagcgcatcg ttcccgccga ccccgccaac cctctctcga acgagcagaa 6420
gatcatggag gccggtcagg tgcgcaacaa cctccacttt gtccgaaacc tccactcgtg 6480
tccaaactcc tcgtcgccaa cttctcctcg acctttctcg tcctcctcga cagctgctga 6540
cctgcttcca ccttgtacca tgcgcaggtc tggtacccga acagctcgtt caagacggga 6600
caggcgatct tcgacttcaa ccgcgagggt ctcccgctca tcatcttcgc caactggcgc 6660
ggcttctcgg gcggccagca ggacatgttt gacgaggtcc tcaagcgcgg ctcgctcatc 6720
gtcgacggtc tctcggcgta caagcagccc gtcttcgtct acatcgtccc gaacggcgaa 6780
cttcgcggcg gtgcttgggt cgtcctcgac ccgtcgatca acgccgaggg catgatggag 6840
atgtacgtcg atgagactgc tcgcgccggt gtcctcgagc ccgagggcat cgtcgagatc 6900
aagctccgca aggacaagct cctcgccctc atggaccgcc tcgacccgac ctaccacgcc 6960
ctccgcgtca agtcgaccga cgtctcgctc tcgcccgccg acgctgcgca ggccaagacc 7020
gagctcgccg cgcgcgagaa gcagctcatg ccgatctacc agcaggtcgc gctccagttc 7080
gccgactcgc acgacaaggc cggccgcatc ctcagcaagg gctgcgcgcg cgaggcgctc 7140
gagtggtcga acgctcgtcg ctacttctac gcccgcctcc gccgccgtgt cgccgaggag 7200
gccgccgtca agcgcctggg cg 7222
<210> 90
<211> 6702
<212> DNA
<213> Rhodosporidium toruloides
<220>
<221> CDS
<222> (1)..(6702)
<400> 90
atg cca ttc tct ggc gag gcg aag gcg gtc aac gga tcg cac tcg gtc 48
Met Pro Phe Ser Gly Glu Ala Lys Ala Val Asn Gly Ser His Ser Val
1 5 10 15
gac gag gcg ccg aag aac ccc aag tac gac cat ggg cgg gtc gta aag 96
Asp Glu Ala Pro Lys Asn Pro Lys Tyr Asp His Gly Arg Val Val Lys
20 25 30
tac ctc ggc ggc aac tcg ctc gaa tct gcg ccc cct tcc aag gtc gcc 144
Tyr Leu Gly Gly Asn Ser Leu Glu Ser Ala Pro Pro Ser Lys Val Ala
35 40 45
gac tgg gtc agg gag cgt ggt gga cac acc gtc atc aca aag atc ctc 192
Asp Trp Val Arg Glu Arg Gly Gly His Thr Val Ile Thr Lys Ile Leu
50 55 60
atc gcc aac aat ggt atc gcc gca gtc aag gag atc cgc tcg gtg cgc 240
Ile Ala Asn Asn Gly Ile Ala Ala Val Lys Glu Ile Arg Ser Val Arg
65 70 75 80
aag tgg gcg tac gag acg ttc gga agc gag cgc gcg atc gag ttt acc 288
Lys Trp Ala Tyr Glu Thr Phe Gly Ser Glu Arg Ala Ile Glu Phe Thr
85 90 95
gtc atg gcg acc ccg gag gac ctc aag gtc aac gca gac tac atc cgc 336
Val Met Ala Thr Pro Glu Asp Leu Lys Val Asn Ala Asp Tyr Ile Arg
100 105 110
atg gcc gat cag tac gtc gag gtt ccc ggt gga acc aac aac aac aac 384
Met Ala Asp Gln Tyr Val Glu Val Pro Gly Gly Thr Asn Asn Asn Asn
115 120 125
tac gcc aac gtc gat gtc atc gtc gat gtt gcc gag cgc gca ggc gtc 432
Tyr Ala Asn Val Asp Val Ile Val Asp Val Ala Glu Arg Ala Gly Val
130 135 140
cac gcc gtc tgg gca gga tgg ggc cac gcc tcc gag aac ccc cgc ctt 480
His Ala Val Trp Ala Gly Trp Gly His Ala Ser Glu Asn Pro Arg Leu
145 150 155 160
ccc gag tcg ctc gcc gcc tcg aag cac aag atc gtc ttc atc ggt cct 528
Pro Glu Ser Leu Ala Ala Ser Lys His Lys Ile Val Phe Ile Gly Pro
165 170 175
ccc ggc tcc gcc atg cgc tcg ctc gga gac aag atc tcg tcg acc atc 576
Pro Gly Ser Ala Met Arg Ser Leu Gly Asp Lys Ile Ser Ser Thr Ile
180 185 190
gtc gcg cag cac gcc cag gtt ccg tgc atg gac tgg tcc ggc cag ggc 624
Val Ala Gln His Ala Gln Val Pro Cys Met Asp Trp Ser Gly Gln Gly
195 200 205
gtc gac caa gtc acc cag tcg ccc gag ggc tac gtt act gtc gcc gac 672
Val Asp Gln Val Thr Gln Ser Pro Glu Gly Tyr Val Thr Val Ala Asp
210 215 220
gac gtc tac cag cag gcc tgt gtg cac gac gcc gac gag ggt ctc gcc 720
Asp Val Tyr Gln Gln Ala Cys Val His Asp Ala Asp Glu Gly Leu Ala
225 230 235 240
cgc gcg tcg agg atc gga tac ccc gtc atg atc aag gcg tcc gag gga 768
Arg Ala Ser Arg Ile Gly Tyr Pro Val Met Ile Lys Ala Ser Glu Gly
245 250 255
gga gga gga aag ggt att cgc aag gtc gag aag gag cag gac ttt aag 816
Gly Gly Gly Lys Gly Ile Arg Lys Val Glu Lys Glu Gln Asp Phe Lys
260 265 270
cag gcc ttc cag gct gtc ctc acc gag gtt ccc ggc tcg ccc gtc ttt 864
Gln Ala Phe Gln Ala Val Leu Thr Glu Val Pro Gly Ser Pro Val Phe
275 280 285
atc atg aag ctc gcc ggc gca gct cgc cac ctc gag gtc cag gtt ctc 912
Ile Met Lys Leu Ala Gly Ala Ala Arg His Leu Glu Val Gln Val Leu
290 295 300
gcc gac cag tac ggc aac gcc atc tcg ctc ttc ggc cgt gac tgc tcg 960
Ala Asp Gln Tyr Gly Asn Ala Ile Ser Leu Phe Gly Arg Asp Cys Ser
305 310 315 320
gtt cag cgt cgc cac cag aag atc atc gaa gag gcg ccc gtc acc atc 1008
Val Gln Arg Arg His Gln Lys Ile Ile Glu Glu Ala Pro Val Thr Ile
325 330 335
gcc aag ccc gac acg ttc gag cag atg gaa aag tcg gcc gtc cgc ctt 1056
Ala Lys Pro Asp Thr Phe Glu Gln Met Glu Lys Ser Ala Val Arg Leu
340 345 350
gcc aag ctc gtc ggc tac gtc tcg gcg ggt acc gtc gag ttc ctc tac 1104
Ala Lys Leu Val Gly Tyr Val Ser Ala Gly Thr Val Glu Phe Leu Tyr
355 360 365
tcg gct gcc gac gac aag ttt gcc ttc ctc gag ctc aac ccg cgt ctc 1152
Ser Ala Ala Asp Asp Lys Phe Ala Phe Leu Glu Leu Asn Pro Arg Leu
370 375 380
cag gtc gag cac ccg acc acc gag atg gtt tcg ggc gtc aac ctt ccc 1200
Gln Val Glu His Pro Thr Thr Glu Met Val Ser Gly Val Asn Leu Pro
385 390 395 400
gcc gcc cag ctc cag gtc gct atg ggt gtt ccc ctc cat cgc atc cgc 1248
Ala Ala Gln Leu Gln Val Ala Met Gly Val Pro Leu His Arg Ile Arg
405 410 415
gac atc cgc acg ctc tac ggc aag gca ccc aac ggc agc agc gag atc 1296
Asp Ile Arg Thr Leu Tyr Gly Lys Ala Pro Asn Gly Ser Ser Glu Ile
420 425 430
gat ttc gac ttc gag aac ccc gag tcg gcc aag acg cag cgc aag ccc 1344
Asp Phe Asp Phe Glu Asn Pro Glu Ser Ala Lys Thr Gln Arg Lys Pro
435 440 445
tcg ccg aag ggt cac gtc gtt gcc gta cgt atc acg gct gag aac cct 1392
Ser Pro Lys Gly His Val Val Ala Val Arg Ile Thr Ala Glu Asn Pro
450 455 460
gac gcc ggc ttc aag ccg tcc atg ggt act ctc caa gag ctc aac ttc 1440
Asp Ala Gly Phe Lys Pro Ser Met Gly Thr Leu Gln Glu Leu Asn Phe
465 470 475 480
cgc tcg agc acg aac gtc tgg ggt tac ttc tcc gtc ggc agc gcc ggt 1488
Arg Ser Ser Thr Asn Val Trp Gly Tyr Phe Ser Val Gly Ser Ala Gly
485 490 495
gga ctg cac gag ttt gcc gac tcg cag ttc ggc cac atc ttt gcg tac 1536
Gly Leu His Glu Phe Ala Asp Ser Gln Phe Gly His Ile Phe Ala Tyr
500 505 510
ggc tcg gac cgt tcc gag tcg cgc aag aac atg gtc gtc gcg ctc aag 1584
Gly Ser Asp Arg Ser Glu Ser Arg Lys Asn Met Val Val Ala Leu Lys
515 520 525
gag ctc tcg att cgc ggt gac ttc cgc acg acc gtc gag tac ctc atc 1632
Glu Leu Ser Ile Arg Gly Asp Phe Arg Thr Thr Val Glu Tyr Leu Ile
530 535 540
aag ctt ctc gag acg gac gcg ttc gag cag aac acg atc acg acc gcg 1680
Lys Leu Leu Glu Thr Asp Ala Phe Glu Gln Asn Thr Ile Thr Thr Ala
545 550 555 560
tgg ctc gac agc ctc atc tcg gct cgc ctg acc gcc gag agg ccc gac 1728
Trp Leu Asp Ser Leu Ile Ser Ala Arg Leu Thr Ala Glu Arg Pro Asp
565 570 575
acg act ctc gcc atc atc tgc ggc gcc gtt acc aag gcc cac ctc gct 1776
Thr Thr Leu Ala Ile Ile Cys Gly Ala Val Thr Lys Ala His Leu Ala
580 585 590
tcc gag gcc aac atc gcc gag tac aag cgc atc ctc gag aag ggt cag 1824
Ser Glu Ala Asn Ile Ala Glu Tyr Lys Arg Ile Leu Glu Lys Gly Gln
595 600 605
agc ccc gcc aag gag ctc ctc gcc acc gtc gtc ccg ctc gag ttc gtc 1872
Ser Pro Ala Lys Glu Leu Leu Ala Thr Val Val Pro Leu Glu Phe Val
610 615 620
ctc gag gac gtc aag tac cgc gcg acc gcc tcg cgc tcg tcg cct tcg 1920
Leu Glu Asp Val Lys Tyr Arg Ala Thr Ala Ser Arg Ser Ser Pro Ser
625 630 635 640
agc tgg tcc atc tac gtc aac ggc tcg aac gtc tcc gtc ggc atc cgc 1968
Ser Trp Ser Ile Tyr Val Asn Gly Ser Asn Val Ser Val Gly Ile Arg
645 650 655
cct ctc gcc gac ggc ggt ctc ctc atc ctc ctt gac ggc cgc tcg tac 2016
Pro Leu Ala Asp Gly Gly Leu Leu Ile Leu Leu Asp Gly Arg Ser Tyr
660 665 670
acc tgc tac gcc aag gag gag gtc ggc gcg ctc cgc ctc tcg atc gac 2064
Thr Cys Tyr Ala Lys Glu Glu Val Gly Ala Leu Arg Leu Ser Ile Asp
675 680 685
tcg agg acc gtc ctc att gct cag gag aac gac ccc acc cag ctt cgc 2112
Ser Arg Thr Val Leu Ile Ala Gln Glu Asn Asp Pro Thr Gln Leu Arg
690 695 700
tcg cct tca ccc ggc aag ctc gtc cgc tac ttc atc gag tcc ggc gag 2160
Ser Pro Ser Pro Gly Lys Leu Val Arg Tyr Phe Ile Glu Ser Gly Glu
705 710 715 720
cac atc tcg aag ggc gag gcg tac gct gag atc gag gtc atg aag atg 2208
His Ile Ser Lys Gly Glu Ala Tyr Ala Glu Ile Glu Val Met Lys Met
725 730 735
atc atg ccc ctc atc gct gcc gag gac ggt atc gcg caa ttc atc aag 2256
Ile Met Pro Leu Ile Ala Ala Glu Asp Gly Ile Ala Gln Phe Ile Lys
740 745 750
cag ccg gga gcg acg ctc gag gcc ggc gac atc ctc ggt atc ttg tcg 2304
Gln Pro Gly Ala Thr Leu Glu Ala Gly Asp Ile Leu Gly Ile Leu Ser
755 760 765
ctc gac gac ccg agc cgc gtc cac cac gcc aag ccg ttc gat ggc cag 2352
Leu Asp Asp Pro Ser Arg Val His His Ala Lys Pro Phe Asp Gly Gln
770 775 780
ctt ccc gcc ctt ggc ttg ccc tcc atc gtc ggc aac aag ccg cac cag 2400
Leu Pro Ala Leu Gly Leu Pro Ser Ile Val Gly Asn Lys Pro His Gln
785 790 795 800
cgc ttc gcc tac ctc aaa gac gtg ctc tca aac atc ctc atg ggc tac 2448
Arg Phe Ala Tyr Leu Lys Asp Val Leu Ser Asn Ile Leu Met Gly Tyr
805 810 815
gac aac cag gcc gtc atg cag tcg agc atc aag gag ctc atc tcg gtt 2496
Asp Asn Gln Ala Val Met Gln Ser Ser Ile Lys Glu Leu Ile Ser Val
820 825 830
ctt cgc aac ccc gag ctc ccc tac ggc gag gcc aac gct gtc ctc tcg 2544
Leu Arg Asn Pro Glu Leu Pro Tyr Gly Glu Ala Asn Ala Val Leu Ser
835 840 845
acg ctt tcg ggt cgc atc ccc gcc aag ctc gag cag acc ctc cgc cag 2592
Thr Leu Ser Gly Arg Ile Pro Ala Lys Leu Glu Gln Thr Leu Arg Gln
850 855 860
tac atc gac cag gct cac gag tct ggc gcc gag ttc ccg tcc gcc aag 2640
Tyr Ile Asp Gln Ala His Glu Ser Gly Ala Glu Phe Pro Ser Ala Lys
865 870 875 880
tgc cgc aag gcg atc gac acg acc ctt gag cag ctc cgc ccc gcc gag 2688
Cys Arg Lys Ala Ile Asp Thr Thr Leu Glu Gln Leu Arg Pro Ala Glu
885 890 895
gcg cag act gtc cgc aac ttc ctc gtc gcg ttc gac gac atc gtc tac 2736
Ala Gln Thr Val Arg Asn Phe Leu Val Ala Phe Asp Asp Ile Val Tyr
900 905 910
cgc tac cgc tcg ggc ctc aag cac cac gag tgg tca acg ctc gcc ggc 2784
Arg Tyr Arg Ser Gly Leu Lys His His Glu Trp Ser Thr Leu Ala Gly
915 920 925
atc ttt gcc gcg tac gcc gag acg gag aag ccg ttc agc ggc aag gac 2832
Ile Phe Ala Ala Tyr Ala Glu Thr Glu Lys Pro Phe Ser Gly Lys Asp
930 935 940
ggc gac gtc gtc ctc gag ctc cgc gac gcc cac cgc gac tcg ctc gac 2880
Gly Asp Val Val Leu Glu Leu Arg Asp Ala His Arg Asp Ser Leu Asp
945 950 955 960
tcg gtc gtc aag atc gtt ctc tcg cac tac aag gct gcc tcg aag aac 2928
Ser Val Val Lys Ile Val Leu Ser His Tyr Lys Ala Ala Ser Lys Asn
965 970 975
tcg ctt gtc ctt gcg ctc ctc gac atc gtc aag gac tcg gac gcg gtt 2976
Ser Leu Val Leu Ala Leu Leu Asp Ile Val Lys Asp Ser Asp Ala Val
980 985 990
ccg ctc atc gag cag gtc gtc agc cct gcg ctc aag gac ctc gcc gac 3024
Pro Leu Ile Glu Gln Val Val Ser Pro Ala Leu Lys Asp Leu Ala Asp
995 1000 1005
ctc gac tcg aag gcc acg act aag gtc gcc ctg aag gcc cgc gag 3069
Leu Asp Ser Lys Ala Thr Thr Lys Val Ala Leu Lys Ala Arg Glu
1010 1015 1020
gtg ctc atc cac atc cag ctc ccc tcg ctc gac gag cgc ctc gga 3114
Val Leu Ile His Ile Gln Leu Pro Ser Leu Asp Glu Arg Leu Gly
1025 1030 1035
cag ctc gag cag att ctc aag gcc tcg gtg acg ccc acc gtt tac 3159
Gln Leu Glu Gln Ile Leu Lys Ala Ser Val Thr Pro Thr Val Tyr
1040 1045 1050
ggc gag ccc ggc cac gac cgc act cct cgc ggt gaa gtc ctt aag 3204
Gly Glu Pro Gly His Asp Arg Thr Pro Arg Gly Glu Val Leu Lys
1055 1060 1065
gac gtc atc gac tcg cgc ttc acc gtc ttt gac gtt ctc ccg agc 3249
Asp Val Ile Asp Ser Arg Phe Thr Val Phe Asp Val Leu Pro Ser
1070 1075 1080
ttc ttc cag cac cag gac cac tgg gtc tcg ctc gcc gcg ctc gac 3294
Phe Phe Gln His Gln Asp His Trp Val Ser Leu Ala Ala Leu Asp
1085 1090 1095
acc tac gtc cgc cgc gcc tac cgc tcg tac aac ctc ctc aac atc 3339
Thr Tyr Val Arg Arg Ala Tyr Arg Ser Tyr Asn Leu Leu Asn Ile
1100 1105 1110
gag cac atc gag gcc gat gcc gcc gag gac gag ccc gcg acg gtt 3384
Glu His Ile Glu Ala Asp Ala Ala Glu Asp Glu Pro Ala Thr Val
1115 1120 1125
gcc tgg tcg ttc cgc atg cgc aag gct gcg tcc gag tct gag ccg 3429
Ala Trp Ser Phe Arg Met Arg Lys Ala Ala Ser Glu Ser Glu Pro
1130 1135 1140
ccc acg ccc acg acc ggc ctc acg tcg cag cgc acc gcc tcg tac 3474
Pro Thr Pro Thr Thr Gly Leu Thr Ser Gln Arg Thr Ala Ser Tyr
1145 1150 1155
tcg gac ttg acg ttc ctc ctc aac aac gcc cag tcc gag ccg atc 3519
Ser Asp Leu Thr Phe Leu Leu Asn Asn Ala Gln Ser Glu Pro Ile
1160 1165 1170
cgc tac ggc gcg atg ttc tcg gtc cgc tcg ctc gac cgc ttc cgc 3564
Arg Tyr Gly Ala Met Phe Ser Val Arg Ser Leu Asp Arg Phe Arg
1175 1180 1185
cag gag ctc ggt acc gtc ctc cga cac ttc ccc gac tcg aac aag 3609
Gln Glu Leu Gly Thr Val Leu Arg His Phe Pro Asp Ser Asn Lys
1190 1195 1200
ggc aag ctc cag cag cag cct gcc gcg tcg tcg agc cag gag cag 3654
Gly Lys Leu Gln Gln Gln Pro Ala Ala Ser Ser Ser Gln Glu Gln
1205 1210 1215
tgg aac gtc atc aac gtc gcg ctc acg gtc ccc gcc agc gcg cag 3699
Trp Asn Val Ile Asn Val Ala Leu Thr Val Pro Ala Ser Ala Gln
1220 1225 1230
gtc gac gag gac gct ctc cgc gcc gac ttt gcc gct cac gtg aac 3744
Val Asp Glu Asp Ala Leu Arg Ala Asp Phe Ala Ala His Val Asn
1235 1240 1245
gcg atg agc gcc gag atc gac gct cgc ggc atg cgc cgc ctc acc 3789
Ala Met Ser Ala Glu Ile Asp Ala Arg Gly Met Arg Arg Leu Thr
1250 1255 1260
ctc ctc atc tgc cgc gag ggc cag tac ccg tcc tac tac acc gtc 3834
Leu Leu Ile Cys Arg Glu Gly Gln Tyr Pro Ser Tyr Tyr Thr Val
1265 1270 1275
cgc aag cag gac ggc acc tgg aag gag ctc gag acg atc cgc gac 3879
Arg Lys Gln Asp Gly Thr Trp Lys Glu Leu Glu Thr Ile Arg Asp
1280 1285 1290
atc gag ccc gcc ctc gcc ttc cag ctc gag ttg ggc cgc ctc tcc 3924
Ile Glu Pro Ala Leu Ala Phe Gln Leu Glu Leu Gly Arg Leu Ser
1295 1300 1305
aac ttc cac ctc gag ccg tgc ccc gtt gag aac cgc cag gtc cac 3969
Asn Phe His Leu Glu Pro Cys Pro Val Glu Asn Arg Gln Val His
1310 1315 1320
gtc tac tac gcg acc gcc aag ggc aac tcg tcc gac tgc cgc ttc 4014
Val Tyr Tyr Ala Thr Ala Lys Gly Asn Ser Ser Asp Cys Arg Phe
1325 1330 1335
ttc gtc cgc gca ctc gtc cgc cct ggc cgt ctc cgc ggt aac atg 4059
Phe Val Arg Ala Leu Val Arg Pro Gly Arg Leu Arg Gly Asn Met
1340 1345 1350
aag acg gcc gac tac ctc gtc tcc gag gct gac cgc ctc gtc acc 4104
Lys Thr Ala Asp Tyr Leu Val Ser Glu Ala Asp Arg Leu Val Thr
1355 1360 1365
gat gtc ctc gac tcg ctc gag gtc gcc agc tcg cag cgc cgc gct 4149
Asp Val Leu Asp Ser Leu Glu Val Ala Ser Ser Gln Arg Arg Ala
1370 1375 1380
gcc gac ggc aac cac atc tcg ctc aac ttc ctg tac tct ctc cgt 4194
Ala Asp Gly Asn His Ile Ser Leu Asn Phe Leu Tyr Ser Leu Arg
1385 1390 1395
ctc gac ttt gac gag gtc cag gct gcc ctc gcc ggc ttc atc gac 4239
Leu Asp Phe Asp Glu Val Gln Ala Ala Leu Ala Gly Phe Ile Asp
1400 1405 1410
cgc cac ggc aag cgc ttc tgg cgt ctc cgc gtc acc ggc gcc gag 4284
Arg His Gly Lys Arg Phe Trp Arg Leu Arg Val Thr Gly Ala Glu
1415 1420 1425
atc cgc atc gtc ctc gag gac gcg cag ggc aac att cag ccc atc 4329
Ile Arg Ile Val Leu Glu Asp Ala Gln Gly Asn Ile Gln Pro Ile
1430 1435 1440
cgc gcc atc atc gag aac gtc tcg ggt ttc gtc gtc aag tac gag 4374
Arg Ala Ile Ile Glu Asn Val Ser Gly Phe Val Val Lys Tyr Glu
1445 1450 1455
gcg tac cgc gag gtc acg acc gac aag ggc cag gtc atc ctc aag 4419
Ala Tyr Arg Glu Val Thr Thr Asp Lys Gly Gln Val Ile Leu Lys
1460 1465 1470
tcg atc ggt ccg cag ggc gcg ttg cac ctt cag ccg gtc aac ttc 4464
Ser Ile Gly Pro Gln Gly Ala Leu His Leu Gln Pro Val Asn Phe
1475 1480 1485
ccc tac ccg acc aag gag tgg ctt cag ccg aag cgc tac aag gcc 4509
Pro Tyr Pro Thr Lys Glu Trp Leu Gln Pro Lys Arg Tyr Lys Ala
1490 1495 1500
cac gtc gtc ggc acg acg tac gtc tac gac ttc ccc gac ctt ttc 4554
His Val Val Gly Thr Thr Tyr Val Tyr Asp Phe Pro Asp Leu Phe
1505 1510 1515
cgc cag gca atc cgc aag cag tgg aag gcg gcc ggc aag act gcg 4599
Arg Gln Ala Ile Arg Lys Gln Trp Lys Ala Ala Gly Lys Thr Ala
1520 1525 1530
ccc gcc gag ctc ctc gtc gcc aag gag ctc gtc ctc gac gag ttc 4644
Pro Ala Glu Leu Leu Val Ala Lys Glu Leu Val Leu Asp Glu Phe
1535 1540 1545
ggc aag cct cag gag gtc gcc cgc ccg cct ggc acc aac aat atc 4689
Gly Lys Pro Gln Glu Val Ala Arg Pro Pro Gly Thr Asn Asn Ile
1550 1555 1560
ggc atg gtc ggc tgg atc tac acg atc ttc acg ccc gaa tac ccc 4734
Gly Met Val Gly Trp Ile Tyr Thr Ile Phe Thr Pro Glu Tyr Pro
1565 1570 1575
tct ggc cgc cgc gtc gtc gtc atc gcg aac gac atc acg ttc aag 4779
Ser Gly Arg Arg Val Val Val Ile Ala Asn Asp Ile Thr Phe Lys
1580 1585 1590
att ggt tcg ttc ggc ccg gag gag gac cgc tac ttc ttc gcc gtc 4824
Ile Gly Ser Phe Gly Pro Glu Glu Asp Arg Tyr Phe Phe Ala Val
1595 1600 1605
acg cag ctc gcg cgc caa ctt ggc ttg ccg cgc gtc tac ctc tcg 4869
Thr Gln Leu Ala Arg Gln Leu Gly Leu Pro Arg Val Tyr Leu Ser
1610 1615 1620
gcc aac tcg ggt gct cgt ctc ggc att gcc gag gag ctc gtc gac 4914
Ala Asn Ser Gly Ala Arg Leu Gly Ile Ala Glu Glu Leu Val Asp
1625 1630 1635
ttg ttc agc gtc gcg tgg gtc gac agc tcg cgg ccg gag aag ggc 4959
Leu Phe Ser Val Ala Trp Val Asp Ser Ser Arg Pro Glu Lys Gly
1640 1645 1650
ttc aag tac ctc tac cta acc gcc gag aag ctc ggc gag ctc aag 5004
Phe Lys Tyr Leu Tyr Leu Thr Ala Glu Lys Leu Gly Glu Leu Lys
1655 1660 1665
aac aag ggc gag aag agc gtc atc acg aag cgc atc gag gac gag 5049
Asn Lys Gly Glu Lys Ser Val Ile Thr Lys Arg Ile Glu Asp Glu
1670 1675 1680
ggc gag acg cgc tac cag atc acc gac atc atc ggc ttg cag gag 5094
Gly Glu Thr Arg Tyr Gln Ile Thr Asp Ile Ile Gly Leu Gln Glu
1685 1690 1695
ggt ctc ggt gtc gag tcg ctc aag ggc tct ggc ctc atc gcc ggt 5139
Gly Leu Gly Val Glu Ser Leu Lys Gly Ser Gly Leu Ile Ala Gly
1700 1705 1710
gag acg tcg cgc gcg tac gac gac atc ttc acg atc acg ctc gtc 5184
Glu Thr Ser Arg Ala Tyr Asp Asp Ile Phe Thr Ile Thr Leu Val
1715 1720 1725
acc gcc cgc tcg gtc ggt atc ggt gcg tac ctc gtc cgc ctc ggc 5229
Thr Ala Arg Ser Val Gly Ile Gly Ala Tyr Leu Val Arg Leu Gly
1730 1735 1740
cag cgt gcc gtc cag gtc gag ggc cag ccg atc atc ctc acc ggt 5274
Gln Arg Ala Val Gln Val Glu Gly Gln Pro Ile Ile Leu Thr Gly
1745 1750 1755
gcc ggc gcg ctc aac aag gtc ctc ggt cgc gag gtg tac tcg tcc 5319
Ala Gly Ala Leu Asn Lys Val Leu Gly Arg Glu Val Tyr Ser Ser
1760 1765 1770
aac ttg cag ctc ggc ggc acg cag atc atg tac aag aac ggt gtc 5364
Asn Leu Gln Leu Gly Gly Thr Gln Ile Met Tyr Lys Asn Gly Val
1775 1780 1785
tcg cac ttg acg gcc gcc aac gac ctc gag ggt gtc ctc agc atc 5409
Ser His Leu Thr Ala Ala Asn Asp Leu Glu Gly Val Leu Ser Ile
1790 1795 1800
gtc cag tgg ctc gcc ttc gtc ccc gag cac cgc ggc gcg cct ctc 5454
Val Gln Trp Leu Ala Phe Val Pro Glu His Arg Gly Ala Pro Leu
1805 1810 1815
ccg atc atg cct tcg ccc gtc gac ccg tgg gac cgc tcg atc gac 5499
Pro Ile Met Pro Ser Pro Val Asp Pro Trp Asp Arg Ser Ile Asp
1820 1825 1830
tac acg ccc atc aag ggc gcg tac gac ccg cgc tgg ttc ctc gcc 5544
Tyr Thr Pro Ile Lys Gly Ala Tyr Asp Pro Arg Trp Phe Leu Ala
1835 1840 1845
ggc aag acg gac gag gcc gac ggt cgc tgg ctc tct ggc ttc ttc 5589
Gly Lys Thr Asp Glu Ala Asp Gly Arg Trp Leu Ser Gly Phe Phe
1850 1855 1860
gac aag ggc tcg ttc cag gag acg ctc tcg ggc tgg gcg cag acc 5634
Asp Lys Gly Ser Phe Gln Glu Thr Leu Ser Gly Trp Ala Gln Thr
1865 1870 1875
gtc gtc gtc ggt cgc gct cgc ctc ggc ggc atc ccc atg ggc gcc 5679
Val Val Val Gly Arg Ala Arg Leu Gly Gly Ile Pro Met Gly Ala
1880 1885 1890
atc gcg gtc gag acc cgc acc atc gag cgc gtc gtg ccc gcc gac 5724
Ile Ala Val Glu Thr Arg Thr Ile Glu Arg Val Val Pro Ala Asp
1895 1900 1905
cct gcc aac cct ctc tcg aac gag cag aag atc atg gag gcc ggt 5769
Pro Ala Asn Pro Leu Ser Asn Glu Gln Lys Ile Met Glu Ala Gly
1910 1915 1920
cag gtc tgg tat ccc aac agc tcg ttc aag acg gga cag gcg atc 5814
Gln Val Trp Tyr Pro Asn Ser Ser Phe Lys Thr Gly Gln Ala Ile
1925 1930 1935
ttc gac ttc aac cgc gag ggt ctc ccg ctc atc atc ttc gcc aac 5859
Phe Asp Phe Asn Arg Glu Gly Leu Pro Leu Ile Ile Phe Ala Asn
1940 1945 1950
tgg cgc ggc ttc tcg ggc ggc cag cag gac atg ttc gac gag gtc 5904
Trp Arg Gly Phe Ser Gly Gly Gln Gln Asp Met Phe Asp Glu Val
1955 1960 1965
ctc aag cgc ggt tcg ctc att gtc gac ggt ctc tcg gcg tac aag 5949
Leu Lys Arg Gly Ser Leu Ile Val Asp Gly Leu Ser Ala Tyr Lys
1970 1975 1980
cag ccc gtc ttc gtc tac atc gtc ccg aac ggc gaa ctt cgc ggc 5994
Gln Pro Val Phe Val Tyr Ile Val Pro Asn Gly Glu Leu Arg Gly
1985 1990 1995
ggt gct tgg gtc gtc ctc gac ccg tcg atc aac gcc gag ggc atg 6039
Gly Ala Trp Val Val Leu Asp Pro Ser Ile Asn Ala Glu Gly Met
2000 2005 2010
atg gag atg tac gtc gac gag act gct cgc gcc ggt gtc ctc gag 6084
Met Glu Met Tyr Val Asp Glu Thr Ala Arg Ala Gly Val Leu Glu
2015 2020 2025
ccc gag ggc atc gtc gag atc aag ctc cgc aag gac aag ctc ctc 6129
Pro Glu Gly Ile Val Glu Ile Lys Leu Arg Lys Asp Lys Leu Leu
2030 2035 2040
gcc ctc atg gac cgc ctc gac ccg acc tac cac gcc ctc cgc gtc 6174
Ala Leu Met Asp Arg Leu Asp Pro Thr Tyr His Ala Leu Arg Val
2045 2050 2055
aag tcg acc gac gct tcg ctc tcg ccc gcc gac gcc gcg cag gcc 6219
Lys Ser Thr Asp Ala Ser Leu Ser Pro Ala Asp Ala Ala Gln Ala
2060 2065 2070
aag acc gag ctc gcc gcg cgc gag aag cag ctc atg ccg atc tac 6264
Lys Thr Glu Leu Ala Ala Arg Glu Lys Gln Leu Met Pro Ile Tyr
2075 2080 2085
cag cag gtc gcg ctc cag ttc gcc gac tcg cac gac aag gcc ggc 6309
Gln Gln Val Ala Leu Gln Phe Ala Asp Ser His Asp Lys Ala Gly
2090 2095 2100
cgc atc ctc agc aag ggc tgc gcg cgc gag gcc ctc gag tgg tcg 6354
Arg Ile Leu Ser Lys Gly Cys Ala Arg Glu Ala Leu Glu Trp Ser
2105 2110 2115
aac gct cgt cgc tac ttc tac gcc cgc ctc cgc cgc cgt ctc gcc 6399
Asn Ala Arg Arg Tyr Phe Tyr Ala Arg Leu Arg Arg Arg Leu Ala
2120 2125 2130
gag gag gcc gcc gtc aag cgt ctc ggc gac gcc gac ccg acc ctc 6444
Glu Glu Ala Ala Val Lys Arg Leu Gly Asp Ala Asp Pro Thr Leu
2135 2140 2145
tcg cgc gac gag cgc ctc gcc atc gtc cac gac gcc gtc ggc cag 6489
Ser Arg Asp Glu Arg Leu Ala Ile Val His Asp Ala Val Gly Gln
2150 2155 2160
ggt gtc gac ctc aac aac gac ctc gct gct gcc gcc gcg ttc gag 6534
Gly Val Asp Leu Asn Asn Asp Leu Ala Ala Ala Ala Ala Phe Glu
2165 2170 2175
cag ggc gcc gcc gcc atc acc gag cgc gtc aag ctc gcg cgc gcg 6579
Gln Gly Ala Ala Ala Ile Thr Glu Arg Val Lys Leu Ala Arg Ala
2180 2185 2190
acg acc gtc gcc tcg act ctc gcg cag ctc gcg cag gac gac aag 6624
Thr Thr Val Ala Ser Thr Leu Ala Gln Leu Ala Gln Asp Asp Lys
2195 2200 2205
gag gct ttc gcc gcc tcg ctc cag cag gtc ctc ggc gac aag ctc 6669
Glu Ala Phe Ala Ala Ser Leu Gln Gln Val Leu Gly Asp Lys Leu
2210 2215 2220
acc gcc gcc gac ctc gcc cgc atc ctc gcc tag 6702
Thr Ala Ala Asp Leu Ala Arg Ile Leu Ala
2225 2230
<210> 91
<211> 2233
<212> PRT
<213> Rhodosporidium toruloides
<400> 91
Met Pro Phe Ser Gly Glu Ala Lys Ala Val Asn Gly Ser His Ser Val
1 5 10 15
Asp Glu Ala Pro Lys Asn Pro Lys Tyr Asp His Gly Arg Val Val Lys
20 25 30
Tyr Leu Gly Gly Asn Ser Leu Glu Ser Ala Pro Pro Ser Lys Val Ala
35 40 45
Asp Trp Val Arg Glu Arg Gly Gly His Thr Val Ile Thr Lys Ile Leu
50 55 60
Ile Ala Asn Asn Gly Ile Ala Ala Val Lys Glu Ile Arg Ser Val Arg
65 70 75 80
Lys Trp Ala Tyr Glu Thr Phe Gly Ser Glu Arg Ala Ile Glu Phe Thr
85 90 95
Val Met Ala Thr Pro Glu Asp Leu Lys Val Asn Ala Asp Tyr Ile Arg
100 105 110
Met Ala Asp Gln Tyr Val Glu Val Pro Gly Gly Thr Asn Asn Asn Asn
115 120 125
Tyr Ala Asn Val Asp Val Ile Val Asp Val Ala Glu Arg Ala Gly Val
130 135 140
His Ala Val Trp Ala Gly Trp Gly His Ala Ser Glu Asn Pro Arg Leu
145 150 155 160
Pro Glu Ser Leu Ala Ala Ser Lys His Lys Ile Val Phe Ile Gly Pro
165 170 175
Pro Gly Ser Ala Met Arg Ser Leu Gly Asp Lys Ile Ser Ser Thr Ile
180 185 190
Val Ala Gln His Ala Gln Val Pro Cys Met Asp Trp Ser Gly Gln Gly
195 200 205
Val Asp Gln Val Thr Gln Ser Pro Glu Gly Tyr Val Thr Val Ala Asp
210 215 220
Asp Val Tyr Gln Gln Ala Cys Val His Asp Ala Asp Glu Gly Leu Ala
225 230 235 240
Arg Ala Ser Arg Ile Gly Tyr Pro Val Met Ile Lys Ala Ser Glu Gly
245 250 255
Gly Gly Gly Lys Gly Ile Arg Lys Val Glu Lys Glu Gln Asp Phe Lys
260 265 270
Gln Ala Phe Gln Ala Val Leu Thr Glu Val Pro Gly Ser Pro Val Phe
275 280 285
Ile Met Lys Leu Ala Gly Ala Ala Arg His Leu Glu Val Gln Val Leu
290 295 300
Ala Asp Gln Tyr Gly Asn Ala Ile Ser Leu Phe Gly Arg Asp Cys Ser
305 310 315 320
Val Gln Arg Arg His Gln Lys Ile Ile Glu Glu Ala Pro Val Thr Ile
325 330 335
Ala Lys Pro Asp Thr Phe Glu Gln Met Glu Lys Ser Ala Val Arg Leu
340 345 350
Ala Lys Leu Val Gly Tyr Val Ser Ala Gly Thr Val Glu Phe Leu Tyr
355 360 365
Ser Ala Ala Asp Asp Lys Phe Ala Phe Leu Glu Leu Asn Pro Arg Leu
370 375 380
Gln Val Glu His Pro Thr Thr Glu Met Val Ser Gly Val Asn Leu Pro
385 390 395 400
Ala Ala Gln Leu Gln Val Ala Met Gly Val Pro Leu His Arg Ile Arg
405 410 415
Asp Ile Arg Thr Leu Tyr Gly Lys Ala Pro Asn Gly Ser Ser Glu Ile
420 425 430
Asp Phe Asp Phe Glu Asn Pro Glu Ser Ala Lys Thr Gln Arg Lys Pro
435 440 445
Ser Pro Lys Gly His Val Val Ala Val Arg Ile Thr Ala Glu Asn Pro
450 455 460
Asp Ala Gly Phe Lys Pro Ser Met Gly Thr Leu Gln Glu Leu Asn Phe
465 470 475 480
Arg Ser Ser Thr Asn Val Trp Gly Tyr Phe Ser Val Gly Ser Ala Gly
485 490 495
Gly Leu His Glu Phe Ala Asp Ser Gln Phe Gly His Ile Phe Ala Tyr
500 505 510
Gly Ser Asp Arg Ser Glu Ser Arg Lys Asn Met Val Val Ala Leu Lys
515 520 525
Glu Leu Ser Ile Arg Gly Asp Phe Arg Thr Thr Val Glu Tyr Leu Ile
530 535 540
Lys Leu Leu Glu Thr Asp Ala Phe Glu Gln Asn Thr Ile Thr Thr Ala
545 550 555 560
Trp Leu Asp Ser Leu Ile Ser Ala Arg Leu Thr Ala Glu Arg Pro Asp
565 570 575
Thr Thr Leu Ala Ile Ile Cys Gly Ala Val Thr Lys Ala His Leu Ala
580 585 590
Ser Glu Ala Asn Ile Ala Glu Tyr Lys Arg Ile Leu Glu Lys Gly Gln
595 600 605
Ser Pro Ala Lys Glu Leu Leu Ala Thr Val Val Pro Leu Glu Phe Val
610 615 620
Leu Glu Asp Val Lys Tyr Arg Ala Thr Ala Ser Arg Ser Ser Pro Ser
625 630 635 640
Ser Trp Ser Ile Tyr Val Asn Gly Ser Asn Val Ser Val Gly Ile Arg
645 650 655
Pro Leu Ala Asp Gly Gly Leu Leu Ile Leu Leu Asp Gly Arg Ser Tyr
660 665 670
Thr Cys Tyr Ala Lys Glu Glu Val Gly Ala Leu Arg Leu Ser Ile Asp
675 680 685
Ser Arg Thr Val Leu Ile Ala Gln Glu Asn Asp Pro Thr Gln Leu Arg
690 695 700
Ser Pro Ser Pro Gly Lys Leu Val Arg Tyr Phe Ile Glu Ser Gly Glu
705 710 715 720
His Ile Ser Lys Gly Glu Ala Tyr Ala Glu Ile Glu Val Met Lys Met
725 730 735
Ile Met Pro Leu Ile Ala Ala Glu Asp Gly Ile Ala Gln Phe Ile Lys
740 745 750
Gln Pro Gly Ala Thr Leu Glu Ala Gly Asp Ile Leu Gly Ile Leu Ser
755 760 765
Leu Asp Asp Pro Ser Arg Val His His Ala Lys Pro Phe Asp Gly Gln
770 775 780
Leu Pro Ala Leu Gly Leu Pro Ser Ile Val Gly Asn Lys Pro His Gln
785 790 795 800
Arg Phe Ala Tyr Leu Lys Asp Val Leu Ser Asn Ile Leu Met Gly Tyr
805 810 815
Asp Asn Gln Ala Val Met Gln Ser Ser Ile Lys Glu Leu Ile Ser Val
820 825 830
Leu Arg Asn Pro Glu Leu Pro Tyr Gly Glu Ala Asn Ala Val Leu Ser
835 840 845
Thr Leu Ser Gly Arg Ile Pro Ala Lys Leu Glu Gln Thr Leu Arg Gln
850 855 860
Tyr Ile Asp Gln Ala His Glu Ser Gly Ala Glu Phe Pro Ser Ala Lys
865 870 875 880
Cys Arg Lys Ala Ile Asp Thr Thr Leu Glu Gln Leu Arg Pro Ala Glu
885 890 895
Ala Gln Thr Val Arg Asn Phe Leu Val Ala Phe Asp Asp Ile Val Tyr
900 905 910
Arg Tyr Arg Ser Gly Leu Lys His His Glu Trp Ser Thr Leu Ala Gly
915 920 925
Ile Phe Ala Ala Tyr Ala Glu Thr Glu Lys Pro Phe Ser Gly Lys Asp
930 935 940
Gly Asp Val Val Leu Glu Leu Arg Asp Ala His Arg Asp Ser Leu Asp
945 950 955 960
Ser Val Val Lys Ile Val Leu Ser His Tyr Lys Ala Ala Ser Lys Asn
965 970 975
Ser Leu Val Leu Ala Leu Leu Asp Ile Val Lys Asp Ser Asp Ala Val
980 985 990
Pro Leu Ile Glu Gln Val Val Ser Pro Ala Leu Lys Asp Leu Ala Asp
995 1000 1005
Leu Asp Ser Lys Ala Thr Thr Lys Val Ala Leu Lys Ala Arg Glu
1010 1015 1020
Val Leu Ile His Ile Gln Leu Pro Ser Leu Asp Glu Arg Leu Gly
1025 1030 1035
Gln Leu Glu Gln Ile Leu Lys Ala Ser Val Thr Pro Thr Val Tyr
1040 1045 1050
Gly Glu Pro Gly His Asp Arg Thr Pro Arg Gly Glu Val Leu Lys
1055 1060 1065
Asp Val Ile Asp Ser Arg Phe Thr Val Phe Asp Val Leu Pro Ser
1070 1075 1080
Phe Phe Gln His Gln Asp His Trp Val Ser Leu Ala Ala Leu Asp
1085 1090 1095
Thr Tyr Val Arg Arg Ala Tyr Arg Ser Tyr Asn Leu Leu Asn Ile
1100 1105 1110
Glu His Ile Glu Ala Asp Ala Ala Glu Asp Glu Pro Ala Thr Val
1115 1120 1125
Ala Trp Ser Phe Arg Met Arg Lys Ala Ala Ser Glu Ser Glu Pro
1130 1135 1140
Pro Thr Pro Thr Thr Gly Leu Thr Ser Gln Arg Thr Ala Ser Tyr
1145 1150 1155
Ser Asp Leu Thr Phe Leu Leu Asn Asn Ala Gln Ser Glu Pro Ile
1160 1165 1170
Arg Tyr Gly Ala Met Phe Ser Val Arg Ser Leu Asp Arg Phe Arg
1175 1180 1185
Gln Glu Leu Gly Thr Val Leu Arg His Phe Pro Asp Ser Asn Lys
1190 1195 1200
Gly Lys Leu Gln Gln Gln Pro Ala Ala Ser Ser Ser Gln Glu Gln
1205 1210 1215
Trp Asn Val Ile Asn Val Ala Leu Thr Val Pro Ala Ser Ala Gln
1220 1225 1230
Val Asp Glu Asp Ala Leu Arg Ala Asp Phe Ala Ala His Val Asn
1235 1240 1245
Ala Met Ser Ala Glu Ile Asp Ala Arg Gly Met Arg Arg Leu Thr
1250 1255 1260
Leu Leu Ile Cys Arg Glu Gly Gln Tyr Pro Ser Tyr Tyr Thr Val
1265 1270 1275
Arg Lys Gln Asp Gly Thr Trp Lys Glu Leu Glu Thr Ile Arg Asp
1280 1285 1290
Ile Glu Pro Ala Leu Ala Phe Gln Leu Glu Leu Gly Arg Leu Ser
1295 1300 1305
Asn Phe His Leu Glu Pro Cys Pro Val Glu Asn Arg Gln Val His
1310 1315 1320
Val Tyr Tyr Ala Thr Ala Lys Gly Asn Ser Ser Asp Cys Arg Phe
1325 1330 1335
Phe Val Arg Ala Leu Val Arg Pro Gly Arg Leu Arg Gly Asn Met
1340 1345 1350
Lys Thr Ala Asp Tyr Leu Val Ser Glu Ala Asp Arg Leu Val Thr
1355 1360 1365
Asp Val Leu Asp Ser Leu Glu Val Ala Ser Ser Gln Arg Arg Ala
1370 1375 1380
Ala Asp Gly Asn His Ile Ser Leu Asn Phe Leu Tyr Ser Leu Arg
1385 1390 1395
Leu Asp Phe Asp Glu Val Gln Ala Ala Leu Ala Gly Phe Ile Asp
1400 1405 1410
Arg His Gly Lys Arg Phe Trp Arg Leu Arg Val Thr Gly Ala Glu
1415 1420 1425
Ile Arg Ile Val Leu Glu Asp Ala Gln Gly Asn Ile Gln Pro Ile
1430 1435 1440
Arg Ala Ile Ile Glu Asn Val Ser Gly Phe Val Val Lys Tyr Glu
1445 1450 1455
Ala Tyr Arg Glu Val Thr Thr Asp Lys Gly Gln Val Ile Leu Lys
1460 1465 1470
Ser Ile Gly Pro Gln Gly Ala Leu His Leu Gln Pro Val Asn Phe
1475 1480 1485
Pro Tyr Pro Thr Lys Glu Trp Leu Gln Pro Lys Arg Tyr Lys Ala
1490 1495 1500
His Val Val Gly Thr Thr Tyr Val Tyr Asp Phe Pro Asp Leu Phe
1505 1510 1515
Arg Gln Ala Ile Arg Lys Gln Trp Lys Ala Ala Gly Lys Thr Ala
1520 1525 1530
Pro Ala Glu Leu Leu Val Ala Lys Glu Leu Val Leu Asp Glu Phe
1535 1540 1545
Gly Lys Pro Gln Glu Val Ala Arg Pro Pro Gly Thr Asn Asn Ile
1550 1555 1560
Gly Met Val Gly Trp Ile Tyr Thr Ile Phe Thr Pro Glu Tyr Pro
1565 1570 1575
Ser Gly Arg Arg Val Val Val Ile Ala Asn Asp Ile Thr Phe Lys
1580 1585 1590
Ile Gly Ser Phe Gly Pro Glu Glu Asp Arg Tyr Phe Phe Ala Val
1595 1600 1605
Thr Gln Leu Ala Arg Gln Leu Gly Leu Pro Arg Val Tyr Leu Ser
1610 1615 1620
Ala Asn Ser Gly Ala Arg Leu Gly Ile Ala Glu Glu Leu Val Asp
1625 1630 1635
Leu Phe Ser Val Ala Trp Val Asp Ser Ser Arg Pro Glu Lys Gly
1640 1645 1650
Phe Lys Tyr Leu Tyr Leu Thr Ala Glu Lys Leu Gly Glu Leu Lys
1655 1660 1665
Asn Lys Gly Glu Lys Ser Val Ile Thr Lys Arg Ile Glu Asp Glu
1670 1675 1680
Gly Glu Thr Arg Tyr Gln Ile Thr Asp Ile Ile Gly Leu Gln Glu
1685 1690 1695
Gly Leu Gly Val Glu Ser Leu Lys Gly Ser Gly Leu Ile Ala Gly
1700 1705 1710
Glu Thr Ser Arg Ala Tyr Asp Asp Ile Phe Thr Ile Thr Leu Val
1715 1720 1725
Thr Ala Arg Ser Val Gly Ile Gly Ala Tyr Leu Val Arg Leu Gly
1730 1735 1740
Gln Arg Ala Val Gln Val Glu Gly Gln Pro Ile Ile Leu Thr Gly
1745 1750 1755
Ala Gly Ala Leu Asn Lys Val Leu Gly Arg Glu Val Tyr Ser Ser
1760 1765 1770
Asn Leu Gln Leu Gly Gly Thr Gln Ile Met Tyr Lys Asn Gly Val
1775 1780 1785
Ser His Leu Thr Ala Ala Asn Asp Leu Glu Gly Val Leu Ser Ile
1790 1795 1800
Val Gln Trp Leu Ala Phe Val Pro Glu His Arg Gly Ala Pro Leu
1805 1810 1815
Pro Ile Met Pro Ser Pro Val Asp Pro Trp Asp Arg Ser Ile Asp
1820 1825 1830
Tyr Thr Pro Ile Lys Gly Ala Tyr Asp Pro Arg Trp Phe Leu Ala
1835 1840 1845
Gly Lys Thr Asp Glu Ala Asp Gly Arg Trp Leu Ser Gly Phe Phe
1850 1855 1860
Asp Lys Gly Ser Phe Gln Glu Thr Leu Ser Gly Trp Ala Gln Thr
1865 1870 1875
Val Val Val Gly Arg Ala Arg Leu Gly Gly Ile Pro Met Gly Ala
1880 1885 1890
Ile Ala Val Glu Thr Arg Thr Ile Glu Arg Val Val Pro Ala Asp
1895 1900 1905
Pro Ala Asn Pro Leu Ser Asn Glu Gln Lys Ile Met Glu Ala Gly
1910 1915 1920
Gln Val Trp Tyr Pro Asn Ser Ser Phe Lys Thr Gly Gln Ala Ile
1925 1930 1935
Phe Asp Phe Asn Arg Glu Gly Leu Pro Leu Ile Ile Phe Ala Asn
1940 1945 1950
Trp Arg Gly Phe Ser Gly Gly Gln Gln Asp Met Phe Asp Glu Val
1955 1960 1965
Leu Lys Arg Gly Ser Leu Ile Val Asp Gly Leu Ser Ala Tyr Lys
1970 1975 1980
Gln Pro Val Phe Val Tyr Ile Val Pro Asn Gly Glu Leu Arg Gly
1985 1990 1995
Gly Ala Trp Val Val Leu Asp Pro Ser Ile Asn Ala Glu Gly Met
2000 2005 2010
Met Glu Met Tyr Val Asp Glu Thr Ala Arg Ala Gly Val Leu Glu
2015 2020 2025
Pro Glu Gly Ile Val Glu Ile Lys Leu Arg Lys Asp Lys Leu Leu
2030 2035 2040
Ala Leu Met Asp Arg Leu Asp Pro Thr Tyr His Ala Leu Arg Val
2045 2050 2055
Lys Ser Thr Asp Ala Ser Leu Ser Pro Ala Asp Ala Ala Gln Ala
2060 2065 2070
Lys Thr Glu Leu Ala Ala Arg Glu Lys Gln Leu Met Pro Ile Tyr
2075 2080 2085
Gln Gln Val Ala Leu Gln Phe Ala Asp Ser His Asp Lys Ala Gly
2090 2095 2100
Arg Ile Leu Ser Lys Gly Cys Ala Arg Glu Ala Leu Glu Trp Ser
2105 2110 2115
Asn Ala Arg Arg Tyr Phe Tyr Ala Arg Leu Arg Arg Arg Leu Ala
2120 2125 2130
Glu Glu Ala Ala Val Lys Arg Leu Gly Asp Ala Asp Pro Thr Leu
2135 2140 2145
Ser Arg Asp Glu Arg Leu Ala Ile Val His Asp Ala Val Gly Gln
2150 2155 2160
Gly Val Asp Leu Asn Asn Asp Leu Ala Ala Ala Ala Ala Phe Glu
2165 2170 2175
Gln Gly Ala Ala Ala Ile Thr Glu Arg Val Lys Leu Ala Arg Ala
2180 2185 2190
Thr Thr Val Ala Ser Thr Leu Ala Gln Leu Ala Gln Asp Asp Lys
2195 2200 2205
Glu Ala Phe Ala Ala Ser Leu Gln Gln Val Leu Gly Asp Lys Leu
2210 2215 2220
Thr Ala Ala Asp Leu Ala Arg Ile Leu Ala
2225 2230
<210> 92
<211> 1925
<212> DNA
<213> Rhodosporidium toruloides
<400> 92
accgcgtcca aggagaagga gcacgatgtc ctcgcctctt ccgactccga ggacgagcac 60
aaccaggacc cgctcaaggc cctcgagaac gagtacccgc cgttcgtcgt gcccaactac 120
tccatcaagg agttgctcgg cgccatcccc gcccactgct tcgagcgcag cgccctccgc 180
tcgtcgctgt acgtcctcgg cgactttgcc atgctcgcgg gtctcgggta cgccgcgtcg 240
cacatcgacc cggccttttc gttcgacggc ggcaaggtgc tcagcggctg ggccggattc 300
gcagccaagt gggccctctg gtccgcctac tgggtcctcg ccggctgggt cggaacgggc 360
gtctggatcc tcggacacga gtgcggccac caggcgttct cgacgtccaa gacgatcaac 420
aacacgatgg gcctcttcct ccactcattc gttctcgtcc cgtaccactc gtggcgcatc 480
tcgcacgcca agcaccacgc cgccacggga cacatgactc gcgacgaggt ctttgtcccg 540
cgcaccgcgt cgttccgcaa ccccaagccc accggcaaga aactccgcgt ctcgcacaac 600
atcgagctcg acgagctcct cgaggatgcg cccctgtacc gcctcggctg gcttctcgtc 660
cagcagctct tcggctggcc cgcgtacctc ttctcgaacg catcgggaca gctctggtac 720
cccaagtgga ccaaccactt cgacccgtcg tcgctcgtct ttgacgcccg ccaccgcggt 780
caggtcctcg tctcggacgc cttcctcgca ggcatggtcg gcctcctcgt cgcctttggc 840
caggtcgtcg gactcgcagg cgtcgtcaag tactacttca ttccttacct ctttgtgtga 900
gtcccgactt ccgatcctcc gaacttgctt tcctcctcct tgatctttcg ccgagttccc 960
gctgactctc ccggacctcc ctgcgcgatc acagcaacca ctggctcgtc atgatcacct 1020
acctccagca cacggacccc tcgctccctc actacaacgc cggcatgtgg aacttccagc 1080
gcggcgcgct ctgcaccatg gaccgcaaca tgctcggccc cgtcggtcct tacctgatgc 1140
acggcatctg cgagacccac ggtgcgttgc agtctcgcgc gaatctcgtt tagttgggcg 1200
gcagtggctg actttgcttc tcctccctct cttcgtccct tctcactccc ttcctccctt 1260
cccacgcttc ctacgacccc tctccacatt ccacacatcg atgtgcacca cccgcagtcg 1320
cgcaccacct ctcgtccaag atcccgcact accacgcgtg ggaggcgaca gaggcgctca 1380
agaacttcct tggcgagcac tacaactaca ccgatgaggg gatgttcagg tcgctctgga 1440
aggcgtatag gcagtgccgg ttcgtcgcag cttctctggg ccttgtttga acctttctag 1500
acgtcgacga tcatattctg acctcccgct cgcctttcct cgcttcacgc agctacgtcg 1560
acgatgaggg cgacgtcctc ttctaccgcg acgcctacgg ccgcgcacgc cgcgtcgccg 1620
tccccgccga ggtcccctcc gactcgggcg tcgagggact ctagacgatg ccctagagcg 1680
agactctttc cttcaccttc cccgcttctc gtagatccct ctctttggtt caagacgcga 1740
ctcctctcgt cgtcgtcggt agatttctcc ttccctctcc ctcctcctca ccactctcct 1800
ctcgagtcgc tcgagtcgtc gttctgttcg gtctgtgccg tgtgccctcc cttcttcgag 1860
tgaagggagg acgcaaaggt ctttgtgtga agcaattatc cactctctac gctcctttta 1920
cacct 1925
<210> 93
<211> 1107
<212> DNA
<213> Rhodosporidium toruloides
<220>
<221> CDS
<222> (1)..(1107)
<400> 93
atg ctc gcg ggt ctc ggg tac gcc gcg tcg cac atc gac ccg gcc ttt 48
Met Leu Ala Gly Leu Gly Tyr Ala Ala Ser His Ile Asp Pro Ala Phe
1 5 10 15
tcg ttc gac ggc ggc aag gtg ctc agc ggc tgg gcc gga ttc gca gcc 96
Ser Phe Asp Gly Gly Lys Val Leu Ser Gly Trp Ala Gly Phe Ala Ala
20 25 30
aag tgg gcc ctc tgg tcc gcc tac tgg gtc ctc gcc ggc tgg gtc gga 144
Lys Trp Ala Leu Trp Ser Ala Tyr Trp Val Leu Ala Gly Trp Val Gly
35 40 45
acg ggc gtc tgg atc ctc gga cac gag tgc ggc cac cag gcg ttc tcg 192
Thr Gly Val Trp Ile Leu Gly His Glu Cys Gly His Gln Ala Phe Ser
50 55 60
acg tcc aag acg atc aac aac acg atg ggc ctc ttc ctc cac tca ttc 240
Thr Ser Lys Thr Ile Asn Asn Thr Met Gly Leu Phe Leu His Ser Phe
65 70 75 80
gtt ctc gtc ccg tac cac tcg tgg cgc atc tcg cac gcc aag cac cac 288
Val Leu Val Pro Tyr His Ser Trp Arg Ile Ser His Ala Lys His His
85 90 95
gcc gcc acg gga cac atg act cgc gac gag gtc ttt gtc ccg cgc acc 336
Ala Ala Thr Gly His Met Thr Arg Asp Glu Val Phe Val Pro Arg Thr
100 105 110
gcg tcg ttc cgc aac ccc aag ccc acc ggc aag aaa ctc cgc gtc tcg 384
Ala Ser Phe Arg Asn Pro Lys Pro Thr Gly Lys Lys Leu Arg Val Ser
115 120 125
cac aac atc gag ctc gac gag ctc ctc gag gat gcg ccc ctg tac cgc 432
His Asn Ile Glu Leu Asp Glu Leu Leu Glu Asp Ala Pro Leu Tyr Arg
130 135 140
ctc ggc tgg ctt ctc gtc cag cag ctc ttc ggc tgg ccc gcg tac ctc 480
Leu Gly Trp Leu Leu Val Gln Gln Leu Phe Gly Trp Pro Ala Tyr Leu
145 150 155 160
ttc tcg aac gca tcg gga cag ctc tgg tac ccc aag tgg acc aac cac 528
Phe Ser Asn Ala Ser Gly Gln Leu Trp Tyr Pro Lys Trp Thr Asn His
165 170 175
ttc gac ccg tcg tcg ctc gtc ttt gac gcc cgc cac cgc ggt cag gtc 576
Phe Asp Pro Ser Ser Leu Val Phe Asp Ala Arg His Arg Gly Gln Val
180 185 190
ctc gtc tcg gac gcc ttc ctc gca ggc atg gtc ggc ctc ctc gtc gcc 624
Leu Val Ser Asp Ala Phe Leu Ala Gly Met Val Gly Leu Leu Val Ala
195 200 205
ttt ggc cag gtc gtc gga ctc gca ggc gtc gtc aag tac tac ttc att 672
Phe Gly Gln Val Val Gly Leu Ala Gly Val Val Lys Tyr Tyr Phe Ile
210 215 220
cct tac ctc ttt gtc aac cac tgg ctc gtc atg atc acc tac ctc cag 720
Pro Tyr Leu Phe Val Asn His Trp Leu Val Met Ile Thr Tyr Leu Gln
225 230 235 240
cac acg gac ccc tcg ctc cct cac tac aac gcc ggc atg tgg aac ttc 768
His Thr Asp Pro Ser Leu Pro His Tyr Asn Ala Gly Met Trp Asn Phe
245 250 255
cag cgc ggc gcg ctc tgc acc atg gac cgc aac atg ctc ggc ccc gtc 816
Gln Arg Gly Ala Leu Cys Thr Met Asp Arg Asn Met Leu Gly Pro Val
260 265 270
ggt cct tac ctg atg cac ggc atc tgc gag acc cac gtc gcg cac cac 864
Gly Pro Tyr Leu Met His Gly Ile Cys Glu Thr His Val Ala His His
275 280 285
ctc tcg tcc aag atc ccg cac tac cac gcg tgg gag gcg aca gag gcg 912
Leu Ser Ser Lys Ile Pro His Tyr His Ala Trp Glu Ala Thr Glu Ala
290 295 300
ctc aag aac ttc ctt ggc gag cac tac aac tac acc gat gag ggg atg 960
Leu Lys Asn Phe Leu Gly Glu His Tyr Asn Tyr Thr Asp Glu Gly Met
305 310 315 320
ttc agg tcg ctc tgg aag gcg tat agg cag tgc cgc tac gtc gac gat 1008
Phe Arg Ser Leu Trp Lys Ala Tyr Arg Gln Cys Arg Tyr Val Asp Asp
325 330 335
gag ggc gac gtc ctc ttc tac cgc gac gcc tac ggc cgc gca cgc cgc 1056
Glu Gly Asp Val Leu Phe Tyr Arg Asp Ala Tyr Gly Arg Ala Arg Arg
340 345 350
gtc gcc gtc ccc gcc gag gtc ccc tcc gac tcg ggc gtc gag gga ctc 1104
Val Ala Val Pro Ala Glu Val Pro Ser Asp Ser Gly Val Glu Gly Leu
355 360 365
tag 1107
<210> 94
<211> 368
<212> PRT
<213> Rhodosporidium toruloides
<400> 94
Met Leu Ala Gly Leu Gly Tyr Ala Ala Ser His Ile Asp Pro Ala Phe
1 5 10 15
Ser Phe Asp Gly Gly Lys Val Leu Ser Gly Trp Ala Gly Phe Ala Ala
20 25 30
Lys Trp Ala Leu Trp Ser Ala Tyr Trp Val Leu Ala Gly Trp Val Gly
35 40 45
Thr Gly Val Trp Ile Leu Gly His Glu Cys Gly His Gln Ala Phe Ser
50 55 60
Thr Ser Lys Thr Ile Asn Asn Thr Met Gly Leu Phe Leu His Ser Phe
65 70 75 80
Val Leu Val Pro Tyr His Ser Trp Arg Ile Ser His Ala Lys His His
85 90 95
Ala Ala Thr Gly His Met Thr Arg Asp Glu Val Phe Val Pro Arg Thr
100 105 110
Ala Ser Phe Arg Asn Pro Lys Pro Thr Gly Lys Lys Leu Arg Val Ser
115 120 125
His Asn Ile Glu Leu Asp Glu Leu Leu Glu Asp Ala Pro Leu Tyr Arg
130 135 140
Leu Gly Trp Leu Leu Val Gln Gln Leu Phe Gly Trp Pro Ala Tyr Leu
145 150 155 160
Phe Ser Asn Ala Ser Gly Gln Leu Trp Tyr Pro Lys Trp Thr Asn His
165 170 175
Phe Asp Pro Ser Ser Leu Val Phe Asp Ala Arg His Arg Gly Gln Val
180 185 190
Leu Val Ser Asp Ala Phe Leu Ala Gly Met Val Gly Leu Leu Val Ala
195 200 205
Phe Gly Gln Val Val Gly Leu Ala Gly Val Val Lys Tyr Tyr Phe Ile
210 215 220
Pro Tyr Leu Phe Val Asn His Trp Leu Val Met Ile Thr Tyr Leu Gln
225 230 235 240
His Thr Asp Pro Ser Leu Pro His Tyr Asn Ala Gly Met Trp Asn Phe
245 250 255
Gln Arg Gly Ala Leu Cys Thr Met Asp Arg Asn Met Leu Gly Pro Val
260 265 270
Gly Pro Tyr Leu Met His Gly Ile Cys Glu Thr His Val Ala His His
275 280 285
Leu Ser Ser Lys Ile Pro His Tyr His Ala Trp Glu Ala Thr Glu Ala
290 295 300
Leu Lys Asn Phe Leu Gly Glu His Tyr Asn Tyr Thr Asp Glu Gly Met
305 310 315 320
Phe Arg Ser Leu Trp Lys Ala Tyr Arg Gln Cys Arg Tyr Val Asp Asp
325 330 335
Glu Gly Asp Val Leu Phe Tyr Arg Asp Ala Tyr Gly Arg Ala Arg Arg
340 345 350
Val Ala Val Pro Ala Glu Val Pro Ser Asp Ser Gly Val Glu Gly Leu
355 360 365
<210> 95
<211> 1371
<212> DNA
<213> Phytophthora infestans
<220>
<221> CDS
<222> (1)..(1371)
<400> 95
atg gtg gac ggc ccc aag acc aag cgc aag att tcg tgg cag gag gtc 48
Met Val Asp Gly Pro Lys Thr Lys Arg Lys Ile Ser Trp Gln Glu Val
1 5 10 15
aag cag cac gcc tcg tac gac aac gcg tgg atc gtc atc cac cac aag 96
Lys Gln His Ala Ser Tyr Asp Asn Ala Trp Ile Val Ile His His Lys
20 25 30
gtc tac gac atc tcc aag tgg gac gcg cac ccc ggc ggc atg gtc atg 144
Val Tyr Asp Ile Ser Lys Trp Asp Ala His Pro Gly Gly Met Val Met
35 40 45
ctc tcg cag gcg ggc gag gac gcg acc gac atc ttc acc gtc tgc cac 192
Leu Ser Gln Ala Gly Glu Asp Ala Thr Asp Ile Phe Thr Val Cys His
50 55 60
ccc acc tcg tcg tgg aag cag ctt gag cag ttc tac atc ggc gac gtg 240
Pro Thr Ser Ser Trp Lys Gln Leu Glu Gln Phe Tyr Ile Gly Asp Val
65 70 75 80
gac gag tcc acc gcg acc gtc aac gag gac ctc tcg gag gag gag aag 288
Asp Glu Ser Thr Ala Thr Val Asn Glu Asp Leu Ser Glu Glu Glu Lys
85 90 95
gcc aag aag gcg aag acc gac gag ttc atc tcg gcg tac cgc cgc ctc 336
Ala Lys Lys Ala Lys Thr Asp Glu Phe Ile Ser Ala Tyr Arg Arg Leu
100 105 110
cgc atc aag atc aag ggc atg ggc ctt tac gac gcg tcg atg gtc ttc 384
Arg Ile Lys Ile Lys Gly Met Gly Leu Tyr Asp Ala Ser Met Val Phe
115 120 125
tac gcg tgg aag atc ctc tcg acc ttc ggc ctc tgg atg gcg tcg gcg 432
Tyr Ala Trp Lys Ile Leu Ser Thr Phe Gly Leu Trp Met Ala Ser Ala
130 135 140
gcg atc tgc tgg cac ttc gac tcg tgg ccc atg tac atg ctt gcg gcg 480
Ala Ile Cys Trp His Phe Asp Ser Trp Pro Met Tyr Met Leu Ala Ala
145 150 155 160
tgc gtc atg gga ctt ttc tgg cag cag tcg ggc tgg ctc gcg cac gac 528
Cys Val Met Gly Leu Phe Trp Gln Gln Ser Gly Trp Leu Ala His Asp
165 170 175
gtg ctc cac cac cag gtc tgg gac aac cac atg atc ggc aac gtc atg 576
Val Leu His His Gln Val Trp Asp Asn His Met Ile Gly Asn Val Met
180 185 190
ggc gtc atc atc gga gac gtc tgg atg ggc ttc tcc gtc cag tgg tgg 624
Gly Val Ile Ile Gly Asp Val Trp Met Gly Phe Ser Val Gln Trp Trp
195 200 205
aag aac aag cac aac ttc cac cac gcg gtc ccc aac ctc atc ggt gac 672
Lys Asn Lys His Asn Phe His His Ala Val Pro Asn Leu Ile Gly Asp
210 215 220
gag aag acg aag tac ctc ggc gac ccc gac atc gac acc atg ccc ctc 720
Glu Lys Thr Lys Tyr Leu Gly Asp Pro Asp Ile Asp Thr Met Pro Leu
225 230 235 240
ctc gcg tgg tcg aag cac atg gcg tcg aag gcg tac gag tcg tcg tgg 768
Leu Ala Trp Ser Lys His Met Ala Ser Lys Ala Tyr Glu Ser Ser Trp
245 250 255
ggc ccc ttc ttc gtc ggc cac cag gcg gtc atc tac ttc ccc ctc ctc 816
Gly Pro Phe Phe Val Gly His Gln Ala Val Ile Tyr Phe Pro Leu Leu
260 265 270
ctc ttc gcg cgc ttc tcg tgg ctc ctc cag tcg tac tac tac gtc ttc 864
Leu Phe Ala Arg Phe Ser Trp Leu Leu Gln Ser Tyr Tyr Tyr Val Phe
275 280 285
aag ggc ttc gcg ttc ggc aag tac gac ccc gtg gac ctc ccc aac ggc 912
Lys Gly Phe Ala Phe Gly Lys Tyr Asp Pro Val Asp Leu Pro Asn Gly
290 295 300
gag aag gtc ggc ctc atg ctc cac tac atc tgg aac gtc atg ctc ccc 960
Glu Lys Val Gly Leu Met Leu His Tyr Ile Trp Asn Val Met Leu Pro
305 310 315 320
gtc gtc acc gga atg tcc gtc gcg cag ggc ctc gcg ttc ttc atg ctt 1008
Val Val Thr Gly Met Ser Val Ala Gln Gly Leu Ala Phe Phe Met Leu
325 330 335
gcg cag atg tcg tgc ggc ggc ttc ctc gcg gcg gtc ttc tcg gtc ggc 1056
Ala Gln Met Ser Cys Gly Gly Phe Leu Ala Ala Val Phe Ser Val Gly
340 345 350
cac aac ggc atg tcg gtc tac gag cgc gag gac aag ccc gac ttc tgg 1104
His Asn Gly Met Ser Val Tyr Glu Arg Glu Asp Lys Pro Asp Phe Trp
355 360 365
cag ctc cag gtc acc acc acc cgc aac atc acc ccc ggc ttc ttc atg 1152
Gln Leu Gln Val Thr Thr Thr Arg Asn Ile Thr Pro Gly Phe Phe Met
370 375 380
gac tgg ttc tgc ggc ggc ctc aac tac cag atc gag cac cac ctc ttc 1200
Asp Trp Phe Cys Gly Gly Leu Asn Tyr Gln Ile Glu His His Leu Phe
385 390 395 400
ccc atg atg ccc cgc cac aac ctc cag aag gtc aac ccc ctc gtc aag 1248
Pro Met Met Pro Arg His Asn Leu Gln Lys Val Asn Pro Leu Val Lys
405 410 415
tcg ctc tgc aag cag tac gac gtg cgc ttc cac gag acg ggc ttc tac 1296
Ser Leu Cys Lys Gln Tyr Asp Val Arg Phe His Glu Thr Gly Phe Tyr
420 425 430
cgg ggc ctc gtc gag gtc gtg gac gag ctt gcg gac atc tcg aag gag 1344
Arg Gly Leu Val Glu Val Val Asp Glu Leu Ala Asp Ile Ser Lys Glu
435 440 445
ttc ctc ctt gag ttc ccc gcg atg tag 1371
Phe Leu Leu Glu Phe Pro Ala Met
450 455
<210> 96
<211> 456
<212> PRT
<213> Phytophthora infestans
<400> 96
Met Val Asp Gly Pro Lys Thr Lys Arg Lys Ile Ser Trp Gln Glu Val
1 5 10 15
Lys Gln His Ala Ser Tyr Asp Asn Ala Trp Ile Val Ile His His Lys
20 25 30
Val Tyr Asp Ile Ser Lys Trp Asp Ala His Pro Gly Gly Met Val Met
35 40 45
Leu Ser Gln Ala Gly Glu Asp Ala Thr Asp Ile Phe Thr Val Cys His
50 55 60
Pro Thr Ser Ser Trp Lys Gln Leu Glu Gln Phe Tyr Ile Gly Asp Val
65 70 75 80
Asp Glu Ser Thr Ala Thr Val Asn Glu Asp Leu Ser Glu Glu Glu Lys
85 90 95
Ala Lys Lys Ala Lys Thr Asp Glu Phe Ile Ser Ala Tyr Arg Arg Leu
100 105 110
Arg Ile Lys Ile Lys Gly Met Gly Leu Tyr Asp Ala Ser Met Val Phe
115 120 125
Tyr Ala Trp Lys Ile Leu Ser Thr Phe Gly Leu Trp Met Ala Ser Ala
130 135 140
Ala Ile Cys Trp His Phe Asp Ser Trp Pro Met Tyr Met Leu Ala Ala
145 150 155 160
Cys Val Met Gly Leu Phe Trp Gln Gln Ser Gly Trp Leu Ala His Asp
165 170 175
Val Leu His His Gln Val Trp Asp Asn His Met Ile Gly Asn Val Met
180 185 190
Gly Val Ile Ile Gly Asp Val Trp Met Gly Phe Ser Val Gln Trp Trp
195 200 205
Lys Asn Lys His Asn Phe His His Ala Val Pro Asn Leu Ile Gly Asp
210 215 220
Glu Lys Thr Lys Tyr Leu Gly Asp Pro Asp Ile Asp Thr Met Pro Leu
225 230 235 240
Leu Ala Trp Ser Lys His Met Ala Ser Lys Ala Tyr Glu Ser Ser Trp
245 250 255
Gly Pro Phe Phe Val Gly His Gln Ala Val Ile Tyr Phe Pro Leu Leu
260 265 270
Leu Phe Ala Arg Phe Ser Trp Leu Leu Gln Ser Tyr Tyr Tyr Val Phe
275 280 285
Lys Gly Phe Ala Phe Gly Lys Tyr Asp Pro Val Asp Leu Pro Asn Gly
290 295 300
Glu Lys Val Gly Leu Met Leu His Tyr Ile Trp Asn Val Met Leu Pro
305 310 315 320
Val Val Thr Gly Met Ser Val Ala Gln Gly Leu Ala Phe Phe Met Leu
325 330 335
Ala Gln Met Ser Cys Gly Gly Phe Leu Ala Ala Val Phe Ser Val Gly
340 345 350
His Asn Gly Met Ser Val Tyr Glu Arg Glu Asp Lys Pro Asp Phe Trp
355 360 365
Gln Leu Gln Val Thr Thr Thr Arg Asn Ile Thr Pro Gly Phe Phe Met
370 375 380
Asp Trp Phe Cys Gly Gly Leu Asn Tyr Gln Ile Glu His His Leu Phe
385 390 395 400
Pro Met Met Pro Arg His Asn Leu Gln Lys Val Asn Pro Leu Val Lys
405 410 415
Ser Leu Cys Lys Gln Tyr Asp Val Arg Phe His Glu Thr Gly Phe Tyr
420 425 430
Arg Gly Leu Val Glu Val Val Asp Glu Leu Ala Asp Ile Ser Lys Glu
435 440 445
Phe Leu Leu Glu Phe Pro Ala Met
450 455
<210> 97
<211> 1374
<212> DNA
<213> Mortierella alpina
<220>
<221> CDS
<222> (1)..(1374)
<400> 97
atg gcg gcg gcg ccc tcg gtc cgc acc ttc acc cgc gcg gag atc ctc 48
Met Ala Ala Ala Pro Ser Val Arg Thr Phe Thr Arg Ala Glu Ile Leu
1 5 10 15
aac gcc gag gcc ctc aac gag ggc aag aag gac gcg gag gcg ccg ttc 96
Asn Ala Glu Ala Leu Asn Glu Gly Lys Lys Asp Ala Glu Ala Pro Phe
20 25 30
ctt atg atc atc gac aac aag gtc tac gac gtg cgc gag ttc gtc ccc 144
Leu Met Ile Ile Asp Asn Lys Val Tyr Asp Val Arg Glu Phe Val Pro
35 40 45
gac cac ccc ggc ggc tcg gtc atc ctc acc cac gtc ggc aag gac ggc 192
Asp His Pro Gly Gly Ser Val Ile Leu Thr His Val Gly Lys Asp Gly
50 55 60
acc gac gtg ttc gac acc ttc cac ccc gag gcg gcg tgg gag acg ctc 240
Thr Asp Val Phe Asp Thr Phe His Pro Glu Ala Ala Trp Glu Thr Leu
65 70 75 80
gcg aac ttc tac gtc ggc gac att gac gag tcc gac cgc gcg atc aag 288
Ala Asn Phe Tyr Val Gly Asp Ile Asp Glu Ser Asp Arg Ala Ile Lys
85 90 95
aac gac gac ttc gcg gcg gag gtc cgc aag ctc cgc acc ctc ttc cag 336
Asn Asp Asp Phe Ala Ala Glu Val Arg Lys Leu Arg Thr Leu Phe Gln
100 105 110
tcg ctc ggc tac tac gac tcg tcg aag gcg tac tac gcg ttc aag gtg 384
Ser Leu Gly Tyr Tyr Asp Ser Ser Lys Ala Tyr Tyr Ala Phe Lys Val
115 120 125
tcg ttc aac ctc tgc atc tgg ggc ctc tcg acc ttc atc gtc gcg aag 432
Ser Phe Asn Leu Cys Ile Trp Gly Leu Ser Thr Phe Ile Val Ala Lys
130 135 140
tgg ggc cag acc tcg acc ctc gcg aac gtc ctc tcg gcc gcg ctc ctc 480
Trp Gly Gln Thr Ser Thr Leu Ala Asn Val Leu Ser Ala Ala Leu Leu
145 150 155 160
ggc ctt ttc tgg cag cag tgc ggc tgg ctc gcc cac gac ttc ctc cac 528
Gly Leu Phe Trp Gln Gln Cys Gly Trp Leu Ala His Asp Phe Leu His
165 170 175
cac cag gtc ttc cag gac cgc ttc tgg ggc gac ctc ttc ggc gcg ttc 576
His Gln Val Phe Gln Asp Arg Phe Trp Gly Asp Leu Phe Gly Ala Phe
180 185 190
ctc ggc ggc gtc tgc cag ggc ttc tcg tcg tcg tgg tgg aag gac aag 624
Leu Gly Gly Val Cys Gln Gly Phe Ser Ser Ser Trp Trp Lys Asp Lys
195 200 205
cac aac acc cac cac gcg gcg ccc aac gtc cac ggc gag gac ccc gac 672
His Asn Thr His His Ala Ala Pro Asn Val His Gly Glu Asp Pro Asp
210 215 220
atc gac acc cac ccc ctc ctc acc tgg tcg gag cac gcg ctt gag atg 720
Ile Asp Thr His Pro Leu Leu Thr Trp Ser Glu His Ala Leu Glu Met
225 230 235 240
ttc tcg gac gtg ccc gac gag gag ctt acc cgc atg tgg tcg cgc ttc 768
Phe Ser Asp Val Pro Asp Glu Glu Leu Thr Arg Met Trp Ser Arg Phe
245 250 255
atg gtc ctc aac cag acc tgg ttc tac ttc ccc atc ctc tcg ttc gcg 816
Met Val Leu Asn Gln Thr Trp Phe Tyr Phe Pro Ile Leu Ser Phe Ala
260 265 270
cgc ctc tcg tgg tgc ctc cag tcc atc atg ttc gtc ctc ccc aac ggc 864
Arg Leu Ser Trp Cys Leu Gln Ser Ile Met Phe Val Leu Pro Asn Gly
275 280 285
cag gcg cac aag ccc tcg ggc gcg cgc gtc ccc atc tcg ctc gtc gag 912
Gln Ala His Lys Pro Ser Gly Ala Arg Val Pro Ile Ser Leu Val Glu
290 295 300
cag ctc tcg ctc gcg atg cac tgg acc tgg tac ttg gcg acc atg ttc 960
Gln Leu Ser Leu Ala Met His Trp Thr Trp Tyr Leu Ala Thr Met Phe
305 310 315 320
ctc ttc atc aag gac ccc gtc aac atg atc gtc tac ttc ctc gtc tcg 1008
Leu Phe Ile Lys Asp Pro Val Asn Met Ile Val Tyr Phe Leu Val Ser
325 330 335
cag gcg gtc tgc ggc aac ctc ctc gcg atc gtc ttc tcc ctc aac cac 1056
Gln Ala Val Cys Gly Asn Leu Leu Ala Ile Val Phe Ser Leu Asn His
340 345 350
aac gga atg ccc gtc atc tcg aag gag gag gcg gtg gac atg gac ttc 1104
Asn Gly Met Pro Val Ile Ser Lys Glu Glu Ala Val Asp Met Asp Phe
355 360 365
ttc acc aag cag atc atc acc ggc cgc gac gtg cac ccc ggc ctc ttc 1152
Phe Thr Lys Gln Ile Ile Thr Gly Arg Asp Val His Pro Gly Leu Phe
370 375 380
gcg gac tgg ttc acc ggc ggc ctc aac tac cag atc gag cac cac ctc 1200
Ala Asp Trp Phe Thr Gly Gly Leu Asn Tyr Gln Ile Glu His His Leu
385 390 395 400
ttc ccc tcg atg ccc cgc cac aac ttc tcg aag atc cag ccc gcc gtc 1248
Phe Pro Ser Met Pro Arg His Asn Phe Ser Lys Ile Gln Pro Ala Val
405 410 415
gag acg ctc tgc aag aag tac ggc gtc cgc tac cac acc acc ggc atg 1296
Glu Thr Leu Cys Lys Lys Tyr Gly Val Arg Tyr His Thr Thr Gly Met
420 425 430
atc gag ggc acc gcc gag gtc ttc tcg cgc ctt aac gag gtg tcc aag 1344
Ile Glu Gly Thr Ala Glu Val Phe Ser Arg Leu Asn Glu Val Ser Lys
435 440 445
gcg gcg tcg aag atg ggc aag gcg cag tag 1374
Ala Ala Ser Lys Met Gly Lys Ala Gln
450 455
<210> 98
<211> 457
<212> PRT
<213> Mortierella alpina
<400> 98
Met Ala Ala Ala Pro Ser Val Arg Thr Phe Thr Arg Ala Glu Ile Leu
1 5 10 15
Asn Ala Glu Ala Leu Asn Glu Gly Lys Lys Asp Ala Glu Ala Pro Phe
20 25 30
Leu Met Ile Ile Asp Asn Lys Val Tyr Asp Val Arg Glu Phe Val Pro
35 40 45
Asp His Pro Gly Gly Ser Val Ile Leu Thr His Val Gly Lys Asp Gly
50 55 60
Thr Asp Val Phe Asp Thr Phe His Pro Glu Ala Ala Trp Glu Thr Leu
65 70 75 80
Ala Asn Phe Tyr Val Gly Asp Ile Asp Glu Ser Asp Arg Ala Ile Lys
85 90 95
Asn Asp Asp Phe Ala Ala Glu Val Arg Lys Leu Arg Thr Leu Phe Gln
100 105 110
Ser Leu Gly Tyr Tyr Asp Ser Ser Lys Ala Tyr Tyr Ala Phe Lys Val
115 120 125
Ser Phe Asn Leu Cys Ile Trp Gly Leu Ser Thr Phe Ile Val Ala Lys
130 135 140
Trp Gly Gln Thr Ser Thr Leu Ala Asn Val Leu Ser Ala Ala Leu Leu
145 150 155 160
Gly Leu Phe Trp Gln Gln Cys Gly Trp Leu Ala His Asp Phe Leu His
165 170 175
His Gln Val Phe Gln Asp Arg Phe Trp Gly Asp Leu Phe Gly Ala Phe
180 185 190
Leu Gly Gly Val Cys Gln Gly Phe Ser Ser Ser Trp Trp Lys Asp Lys
195 200 205
His Asn Thr His His Ala Ala Pro Asn Val His Gly Glu Asp Pro Asp
210 215 220
Ile Asp Thr His Pro Leu Leu Thr Trp Ser Glu His Ala Leu Glu Met
225 230 235 240
Phe Ser Asp Val Pro Asp Glu Glu Leu Thr Arg Met Trp Ser Arg Phe
245 250 255
Met Val Leu Asn Gln Thr Trp Phe Tyr Phe Pro Ile Leu Ser Phe Ala
260 265 270
Arg Leu Ser Trp Cys Leu Gln Ser Ile Met Phe Val Leu Pro Asn Gly
275 280 285
Gln Ala His Lys Pro Ser Gly Ala Arg Val Pro Ile Ser Leu Val Glu
290 295 300
Gln Leu Ser Leu Ala Met His Trp Thr Trp Tyr Leu Ala Thr Met Phe
305 310 315 320
Leu Phe Ile Lys Asp Pro Val Asn Met Ile Val Tyr Phe Leu Val Ser
325 330 335
Gln Ala Val Cys Gly Asn Leu Leu Ala Ile Val Phe Ser Leu Asn His
340 345 350
Asn Gly Met Pro Val Ile Ser Lys Glu Glu Ala Val Asp Met Asp Phe
355 360 365
Phe Thr Lys Gln Ile Ile Thr Gly Arg Asp Val His Pro Gly Leu Phe
370 375 380
Ala Asp Trp Phe Thr Gly Gly Leu Asn Tyr Gln Ile Glu His His Leu
385 390 395 400
Phe Pro Ser Met Pro Arg His Asn Phe Ser Lys Ile Gln Pro Ala Val
405 410 415
Glu Thr Leu Cys Lys Lys Tyr Gly Val Arg Tyr His Thr Thr Gly Met
420 425 430
Ile Glu Gly Thr Ala Glu Val Phe Ser Arg Leu Asn Glu Val Ser Lys
435 440 445
Ala Ala Ser Lys Met Gly Lys Ala Gln
450 455
<210> 99
<211> 1408
<212> DNA
<213> Rhodosporidium toruloides
<400> 99
atgacctcgt acgccgccca tcctcgcgcg tcgtccttcc tcgcctcgtt cgcagacggt 60
cccaagccac caacaccgac aggcatcccc gcgcccctcg cttcgacgta cgacttgttc 120
ttgaaccccg tcacgcccct cgctttcgga ctcgtctact tcgcgacggc caaaaccctc 180
tcccacttcc agaacggcaa gaaccgcatc aagggcaagg gctgggacgt cgctgtcctc 240
gtgcacaata tcctccttgc ggtctactcg gcgtggacgt tcctcggaac cgcgccgcag 300
atcttcggcg ctttcgttcg cggctacatg gcggacgggt tcgccggatt gacccacgcg 360
tgagttccgc gcgcttttac attcgagcgt catctctttg caggaggggc gtcgaattct 420
gaaagacgag cggcgctaac tcgctcgcca actccgacag cttctgcgac tcgtcgttcg 480
cgatctggca gcagacgacg ttccccaagt tcgcctacct cttctacgtc agcaagttct 540
acgtgcgtct ctcctcgctc catctcatag cagtcactca ctcggctcgc tcgcgcagga 600
aatcgtcgac acggccatct tgctgttgaa gggcaagaag gtcggcatgc tccagtcgta 660
ccaccacatg ggcgcgatct ggactatgta cgccgcgtac gctacgcaag ccatgcctgt 720
cgtgcgtccc gcttcctccc tcgctcgaaa acggagaact ctcgctgacg ctcgactccc 780
gcgcgtgcag tggctcttcg tcgtcttcaa ctcgttcatc cactcaatca tgtacaccta 840
ctacgccttc tcgaccgtct cgctcccctt cccgcgcttc ctcaagaaat ccctcacgcg 900
actccaaatc acccagttcc tcgtcggcgg ttcgctcgcc gcctcgtacc tcttcattaa 960
gcttcccgaa ctcccttcgg cggaggagat gtccgctgcg gcgacttcga gcttcgaggc 1020
gggcgtcggc gcgctcaagc gcgagggtcc gacgtgcctc gtcaatgcgg cgcagaggca 1080
tgcgacgttg ctcaattgcg cgtacctcgt gccgttgacg tacctctttg tcgcattctt 1140
cttcaagacg taccagaaga actcggcggc taacgccgcg gcaaaggcca aggccaacgc 1200
gaagaaggcc aactagcgcg gtcccttcct ccctctcttt cgctcctttc gcatccgttt 1260
caactgtcgt gatttcgcac cctcttcgct tgcatacctc tccttagccc tcgtcgtttc 1320
gtcgcagcta gatctcctct cctctctttc tcgtgtccgt attcaccagc tctcttctct 1380
tgggtctctc gtcgtaatcc atagcctc 1408
<210> 100
<211> 990
<212> DNA
<213> Rhodosporidium toruloides
<220>
<221> CDS
<222> (1)..(990)
<400> 100
atg acc tcg tac gcc gcc cat cct cgc gcg tcg tcc ttc ctc gcc tcg 48
Met Thr Ser Tyr Ala Ala His Pro Arg Ala Ser Ser Phe Leu Ala Ser
1 5 10 15
ttc gca gac ggt ccc aag cca cca aca ccg aca ggc atc ccc gcg ccc 96
Phe Ala Asp Gly Pro Lys Pro Pro Thr Pro Thr Gly Ile Pro Ala Pro
20 25 30
ctc gct tcg acg tac gac ttg ttc ttg aac ccc gtc acg ccc ctc gct 144
Leu Ala Ser Thr Tyr Asp Leu Phe Leu Asn Pro Val Thr Pro Leu Ala
35 40 45
ttc gga ctc gtc tac ttc gcg acg gcc aaa acc ctc tcc cac ttc cag 192
Phe Gly Leu Val Tyr Phe Ala Thr Ala Lys Thr Leu Ser His Phe Gln
50 55 60
aac ggc aag aac cgc atc aag ggc aag ggc tgg gac gtc gct gtc ctc 240
Asn Gly Lys Asn Arg Ile Lys Gly Lys Gly Trp Asp Val Ala Val Leu
65 70 75 80
gtg cac aat atc ctc ctt gcg gtc tac tcg gcg tgg acg ttc ctc gga 288
Val His Asn Ile Leu Leu Ala Val Tyr Ser Ala Trp Thr Phe Leu Gly
85 90 95
acc gcg ccg cag atc ttc ggc gct ttc gtt cgc ggc tac atg gcg gac 336
Thr Ala Pro Gln Ile Phe Gly Ala Phe Val Arg Gly Tyr Met Ala Asp
100 105 110
ggg ttc gcc gga ttg acc cac gcc ttc tgc gac tcg tcg ttc gcg atc 384
Gly Phe Ala Gly Leu Thr His Ala Phe Cys Asp Ser Ser Phe Ala Ile
115 120 125
tgg cag cag acg acg ttc ccc aag ttc gcc tac ctc ttc tac gtc agc 432
Trp Gln Gln Thr Thr Phe Pro Lys Phe Ala Tyr Leu Phe Tyr Val Ser
130 135 140
aag ttc tac gaa atc gtc gac acg gcc atc ttg ctg ttg aag ggc aag 480
Lys Phe Tyr Glu Ile Val Asp Thr Ala Ile Leu Leu Leu Lys Gly Lys
145 150 155 160
aag gtc ggc atg ctc cag tcg tac cac cac atg ggc gcg atc tgg act 528
Lys Val Gly Met Leu Gln Ser Tyr His His Met Gly Ala Ile Trp Thr
165 170 175
atg tac gcc gcg tac gct acg caa gcc atg cct gtc tgg ctc ttc gtc 576
Met Tyr Ala Ala Tyr Ala Thr Gln Ala Met Pro Val Trp Leu Phe Val
180 185 190
gtc ttc aac tcg ttc atc cac tca atc atg tac acc tac tac gcc ttc 624
Val Phe Asn Ser Phe Ile His Ser Ile Met Tyr Thr Tyr Tyr Ala Phe
195 200 205
tcg acc gtc tcg ctc ccc ttc ccg cgc ttc ctc aag aaa tcc ctc acg 672
Ser Thr Val Ser Leu Pro Phe Pro Arg Phe Leu Lys Lys Ser Leu Thr
210 215 220
cga ctc caa atc acc cag ttc ctc gtc ggc ggt tcg ctc gcc gcc tcg 720
Arg Leu Gln Ile Thr Gln Phe Leu Val Gly Gly Ser Leu Ala Ala Ser
225 230 235 240
tac ctc ttc att aag ctt ccc gaa ctc cct tcg gcg gag gag atg tcc 768
Tyr Leu Phe Ile Lys Leu Pro Glu Leu Pro Ser Ala Glu Glu Met Ser
245 250 255
gct gcg gcg act tcg agc ttc gag gcg ggc gtc ggc gcg ctc aag cgc 816
Ala Ala Ala Thr Ser Ser Phe Glu Ala Gly Val Gly Ala Leu Lys Arg
260 265 270
gag ggt ccg acg tgc ctc gtc aat gcg gcg cag agg cat gcg acg ttg 864
Glu Gly Pro Thr Cys Leu Val Asn Ala Ala Gln Arg His Ala Thr Leu
275 280 285
ctc aat tgc gcg tac ctc gtg ccg ttg acg tac ctc ttt gtc gca ttc 912
Leu Asn Cys Ala Tyr Leu Val Pro Leu Thr Tyr Leu Phe Val Ala Phe
290 295 300
ttc ttc aag acg tac cag aag aac tcg gcg gct aac gcc gcg gca aag 960
Phe Phe Lys Thr Tyr Gln Lys Asn Ser Ala Ala Asn Ala Ala Ala Lys
305 310 315 320
gcc aag gcc aac gcg aag aag gcc aac tag 990
Ala Lys Ala Asn Ala Lys Lys Ala Asn
325
<210> 101
<211> 329
<212> PRT
<213> Rhodosporidium toruloides
<400> 101
Met Thr Ser Tyr Ala Ala His Pro Arg Ala Ser Ser Phe Leu Ala Ser
1 5 10 15
Phe Ala Asp Gly Pro Lys Pro Pro Thr Pro Thr Gly Ile Pro Ala Pro
20 25 30
Leu Ala Ser Thr Tyr Asp Leu Phe Leu Asn Pro Val Thr Pro Leu Ala
35 40 45
Phe Gly Leu Val Tyr Phe Ala Thr Ala Lys Thr Leu Ser His Phe Gln
50 55 60
Asn Gly Lys Asn Arg Ile Lys Gly Lys Gly Trp Asp Val Ala Val Leu
65 70 75 80
Val His Asn Ile Leu Leu Ala Val Tyr Ser Ala Trp Thr Phe Leu Gly
85 90 95
Thr Ala Pro Gln Ile Phe Gly Ala Phe Val Arg Gly Tyr Met Ala Asp
100 105 110
Gly Phe Ala Gly Leu Thr His Ala Phe Cys Asp Ser Ser Phe Ala Ile
115 120 125
Trp Gln Gln Thr Thr Phe Pro Lys Phe Ala Tyr Leu Phe Tyr Val Ser
130 135 140
Lys Phe Tyr Glu Ile Val Asp Thr Ala Ile Leu Leu Leu Lys Gly Lys
145 150 155 160
Lys Val Gly Met Leu Gln Ser Tyr His His Met Gly Ala Ile Trp Thr
165 170 175
Met Tyr Ala Ala Tyr Ala Thr Gln Ala Met Pro Val Trp Leu Phe Val
180 185 190
Val Phe Asn Ser Phe Ile His Ser Ile Met Tyr Thr Tyr Tyr Ala Phe
195 200 205
Ser Thr Val Ser Leu Pro Phe Pro Arg Phe Leu Lys Lys Ser Leu Thr
210 215 220
Arg Leu Gln Ile Thr Gln Phe Leu Val Gly Gly Ser Leu Ala Ala Ser
225 230 235 240
Tyr Leu Phe Ile Lys Leu Pro Glu Leu Pro Ser Ala Glu Glu Met Ser
245 250 255
Ala Ala Ala Thr Ser Ser Phe Glu Ala Gly Val Gly Ala Leu Lys Arg
260 265 270
Glu Gly Pro Thr Cys Leu Val Asn Ala Ala Gln Arg His Ala Thr Leu
275 280 285
Leu Asn Cys Ala Tyr Leu Val Pro Leu Thr Tyr Leu Phe Val Ala Phe
290 295 300
Phe Phe Lys Thr Tyr Gln Lys Asn Ser Ala Ala Asn Ala Ala Ala Lys
305 310 315 320
Ala Lys Ala Asn Ala Lys Lys Ala Asn
325
<210> 102
<211> 1620
<212> DNA
<213> Rhodosporidium toruloides
<400> 102
ttagttggtc ttcttggcga cggtcgcggc ggcgcccttt gcggcgtcga ccttgttctg 60
gccggccttc ttgtacgtgc gttggtaaaa tgcgataaag aggaagaggt aactggctgt 120
tttcgccagc ccctcgggcg tcagccttgc tcgtagaaaa aaccgagagg aaaggggaaa 180
atggacgacg aggaggaggg gggaaggact cactgaggag tgcgcagccg aagatcgcag 240
ctccctccgt cccagcgcac gacccgaggg tcgggagcga cccgcgggcg tacgtccagg 300
cgaagtagct gtagctggcg aagtagaccg tgaagaggtc gatcacgaac tgggtgattt 360
ggagcgtcgt gaggtaccta taccacgggg gctcgtcagg tctagcacaa agggagagga 420
ggggagagac ggacttcttc caccagatct tgtagccggc ggccgtcata aagtagtagt 480
agtactttac gacggcccac aagcggcgtc agcttccgcg tcgactccag gatgacagga 540
ctcgccgcaa cgcaccatga ggacatgcac aaagaggttg agggtgatga cgacccacga 600
gacggaggtg cgtccattga gctgcgtgta gcagaggacg gcggtcgcgg tgtggtggaa 660
gacgtggagg aactgaaggg gcttcttctt gacgacgagg aagaccgtgt cgaacaactc 720
ccagtctgcg cgcgaggttg cgttaggttg cgcaagcagg ggggcgtgag aaggtcggag 780
agggcgtaca cttgaactgc gcggcgttac aacgtcagtt tcggcatctt cgagggacag 840
agggacaaca cacgtagtag ttgaagatgt agtaggtctc gaggcggggc gtccaggcct 900
cgttggcgca gatggcgtgg aagaggccgt gcttccagat gatggggacg atctgggcgc 960
gggagagcgt aagcgacgtc ctccaaagaa tcagatcgtc gaacgcacct cctcaagcat 1020
cagggcgaga agaaggccgg agccggtcga gaggaggaag ttgtgcagca tgaaaagggg 1080
cttgaagcct gtgcgtcgca ggacgaggtc caagtcagtt tctgtagact ctgcaagctc 1140
aagactttgc tggtgaagct tctgtgcgac aggcgagccc atccgctcga cggtgtcatg 1200
acctcgagcg gttgaagcgt gatcggaacg cgttctcctc gtctatccgg ccttttgctg 1260
ccggtccgag gctcttcagc ctgccagcgt cccattgacg cgtctgcatc tctgatccga 1320
cgccgccgac gcgttgtccc tgcacgagcc ccaacttctt gtcgaagcaa accccaaact 1380
cacggtacgg ctttgatccg cgcatgagcg cctgacctcc gaagatgacc gcgaggtaga 1440
ccgcaacggc aatgacgacc tcgggcgtgg tcgagagggg tgtttggccc gggatccagt 1500
gctcgatggt gcgcgggagg gaggggatgg ggagcgcgtg caggagcgag tagacggggc 1560
cgggggacgg tgcgaccata gtgaatggac ctgagcgggt cgagagcgta gcctgggtgc 1620
<210> 103
<211> 879
<212> DNA
<213> Rhodosporidium toruloides
<220>
<221> CDS
<222> (1)..(879)
<400> 103
atg gtc gca ccg tcc ccc ggc ccc gtc tac tcg ctc ctg cac gcg ctc 48
Met Val Ala Pro Ser Pro Gly Pro Val Tyr Ser Leu Leu His Ala Leu
1 5 10 15
ccc atc ccc tcc ctc ccg cgc acc atc gag cac tgg atc ccg ggc caa 96
Pro Ile Pro Ser Leu Pro Arg Thr Ile Glu His Trp Ile Pro Gly Gln
20 25 30
aca ccc ctc tcg acc acg ccc gag gtc gtc att gcc gtt gcg gtc tac 144
Thr Pro Leu Ser Thr Thr Pro Glu Val Val Ile Ala Val Ala Val Tyr
35 40 45
ctc gcg gtc atc ttc gga ggt cag gcg ctc atg cgc gga tca aag ccg 192
Leu Ala Val Ile Phe Gly Gly Gln Ala Leu Met Arg Gly Ser Lys Pro
50 55 60
tac cgc ttc aag ccc ctt ttc atg ctg cac aac ttc ctc ctc tcg acc 240
Tyr Arg Phe Lys Pro Leu Phe Met Leu His Asn Phe Leu Leu Ser Thr
65 70 75 80
ggc tcc ggc ctt ctt ctc gcc ctg atg ctt gag gag atc gtc ccc atc 288
Gly Ser Gly Leu Leu Leu Ala Leu Met Leu Glu Glu Ile Val Pro Ile
85 90 95
atc tgg aag cac ggc ctc ttc cac gcc atc tgc gcc aac gag gcc tgg 336
Ile Trp Lys His Gly Leu Phe His Ala Ile Cys Ala Asn Glu Ala Trp
100 105 110
acg ccc cgc ctc gag acc tac tac atc ttc aac tac tac ttc aag tac 384
Thr Pro Arg Leu Glu Thr Tyr Tyr Ile Phe Asn Tyr Tyr Phe Lys Tyr
115 120 125
tgg gag ttg ttc gac acg gtc ttc ctc gtc gtc aag aag aag ccc ctt 432
Trp Glu Leu Phe Asp Thr Val Phe Leu Val Val Lys Lys Lys Pro Leu
130 135 140
cag ttc ctc cac gtc ttc cac cac acc gcg acc gcc gtc ctc tgc tac 480
Gln Phe Leu His Val Phe His His Thr Ala Thr Ala Val Leu Cys Tyr
145 150 155 160
acg cag ctc aat gga cgc acc tcc gtc tcg tgg gtc gtc atc acc ctc 528
Thr Gln Leu Asn Gly Arg Thr Ser Val Ser Trp Val Val Ile Thr Leu
165 170 175
aac ctc ttt gtg cat gtc ctc atg tac tac tac tac ttt atg acg gcc 576
Asn Leu Phe Val His Val Leu Met Tyr Tyr Tyr Tyr Phe Met Thr Ala
180 185 190
gcc ggc tac aag atc tgg tgg aag aag tac ctc acg acg ctc caa atc 624
Ala Gly Tyr Lys Ile Trp Trp Lys Lys Tyr Leu Thr Thr Leu Gln Ile
195 200 205
acc cag ttc gtg atc gac ctc ttc acg gtc tac ttc gcc agc tac agc 672
Thr Gln Phe Val Ile Asp Leu Phe Thr Val Tyr Phe Ala Ser Tyr Ser
210 215 220
tac ttc gcc tgg acg tac gcc cgc ggg tcg ctc ccg acc ctc ggg tcg 720
Tyr Phe Ala Trp Thr Tyr Ala Arg Gly Ser Leu Pro Thr Leu Gly Ser
225 230 235 240
tgc gct ggg acg gag gga gct gcg atc ttc ggc tgc gca ctc ctc acc 768
Cys Ala Gly Thr Glu Gly Ala Ala Ile Phe Gly Cys Ala Leu Leu Thr
245 250 255
agt tac ctc ttc ctc ttt atc gca ttt tac caa cgc acg tac aag aag 816
Ser Tyr Leu Phe Leu Phe Ile Ala Phe Tyr Gln Arg Thr Tyr Lys Lys
260 265 270
gcc ggc cag aac aag gtc gac gcc gca aag ggc gcc gcc gcg acc gtc 864
Ala Gly Gln Asn Lys Val Asp Ala Ala Lys Gly Ala Ala Ala Thr Val
275 280 285
gcc aag aag acc aac 879
Ala Lys Lys Thr Asn
290
<210> 104
<211> 293
<212> PRT
<213> Rhodosporidium toruloides
<400> 104
Met Val Ala Pro Ser Pro Gly Pro Val Tyr Ser Leu Leu His Ala Leu
1 5 10 15
Pro Ile Pro Ser Leu Pro Arg Thr Ile Glu His Trp Ile Pro Gly Gln
20 25 30
Thr Pro Leu Ser Thr Thr Pro Glu Val Val Ile Ala Val Ala Val Tyr
35 40 45
Leu Ala Val Ile Phe Gly Gly Gln Ala Leu Met Arg Gly Ser Lys Pro
50 55 60
Tyr Arg Phe Lys Pro Leu Phe Met Leu His Asn Phe Leu Leu Ser Thr
65 70 75 80
Gly Ser Gly Leu Leu Leu Ala Leu Met Leu Glu Glu Ile Val Pro Ile
85 90 95
Ile Trp Lys His Gly Leu Phe His Ala Ile Cys Ala Asn Glu Ala Trp
100 105 110
Thr Pro Arg Leu Glu Thr Tyr Tyr Ile Phe Asn Tyr Tyr Phe Lys Tyr
115 120 125
Trp Glu Leu Phe Asp Thr Val Phe Leu Val Val Lys Lys Lys Pro Leu
130 135 140
Gln Phe Leu His Val Phe His His Thr Ala Thr Ala Val Leu Cys Tyr
145 150 155 160
Thr Gln Leu Asn Gly Arg Thr Ser Val Ser Trp Val Val Ile Thr Leu
165 170 175
Asn Leu Phe Val His Val Leu Met Tyr Tyr Tyr Tyr Phe Met Thr Ala
180 185 190
Ala Gly Tyr Lys Ile Trp Trp Lys Lys Tyr Leu Thr Thr Leu Gln Ile
195 200 205
Thr Gln Phe Val Ile Asp Leu Phe Thr Val Tyr Phe Ala Ser Tyr Ser
210 215 220
Tyr Phe Ala Trp Thr Tyr Ala Arg Gly Ser Leu Pro Thr Leu Gly Ser
225 230 235 240
Cys Ala Gly Thr Glu Gly Ala Ala Ile Phe Gly Cys Ala Leu Leu Thr
245 250 255
Ser Tyr Leu Phe Leu Phe Ile Ala Phe Tyr Gln Arg Thr Tyr Lys Lys
260 265 270
Ala Gly Gln Asn Lys Val Asp Ala Ala Lys Gly Ala Ala Ala Thr Val
275 280 285
Ala Lys Lys Thr Asn
290
<210> 105
<211> 527
<212> PRT
<213> Agaricus bisporus
<400> 105
Met Val Asp Leu Lys Tyr Thr Pro Val Glu Glu Ile Glu Lys Ile Gln
1 5 10 15
Ala Thr Leu Arg Asn Gly Phe Arg Ser Gly Arg Thr Lys Asn Ile Glu
20 25 30
Tyr Arg Lys Tyr Gln Leu Leu Gln Leu Ala Tyr Met Leu Gln Asp Asn
35 40 45
Val Lys Arg Leu Glu Glu Ala Leu Ala Ala Asp Leu Gly Arg Pro Pro
50 55 60
Leu Glu Ser Gln Phe Leu Glu Ile Gly Pro Ser Met Met Asp Ala Arg
65 70 75 80
Asn Ala Trp Ala Gly Val Asp Lys Trp Ala Lys Thr Glu Arg Ala Pro
85 90 95
Phe Ser Ile Asn Gly Phe Ala Met Arg Pro Val Ile Tyr Lys Glu Pro
100 105 110
Lys Gly Val Val Leu Ile Ile Ser Pro Phe Asn Tyr Pro Val Trp Leu
115 120 125
Cys Met Ser Pro Leu Ala Gly Ala Ile Ala Ala Gly Asn Ala Val Leu
130 135 140
Leu Lys Pro Ser Glu Ser Thr Pro His Val Ser Ser Leu Phe Ala Glu
145 150 155 160
Leu Ile Pro Lys Tyr Leu Asp Pro Glu Leu Val Ala Val Val Asn Gly
165 170 175
Gly Val Pro Glu Thr Thr Lys Leu Leu Asp Leu Pro Trp Asp His Ile
180 185 190
Leu Tyr Thr Gly Ser Gly Gln Val Gly Arg Ile Val Ser Ala Ala Ala
195 200 205
Ala Lys His Leu Thr Pro Val Ser Leu Glu Leu Gly Gly Lys Ser Pro
210 215 220
Val Phe Ile Asp Pro Asn Cys Asp Ile Glu Leu Ala Ala Lys Arg Ile
225 230 235 240
Leu Trp Gly Lys Cys Val Asn Ala Gly Gln Thr Cys Thr Ala Pro Asp
245 250 255
Tyr Val Leu Val Pro Arg Glu Val Gln Asp Lys Phe Val Asn Ala Leu
260 265 270
Lys Asn Ser Met Asp Asn Phe Tyr Pro Glu Ser Val Ala Thr Pro Gly
275 280 285
Val Phe Ser Arg Leu Val Thr Pro Gln Ala Phe Asn Arg Ile Lys Gly
290 295 300
Leu Leu Asp Asn Thr Lys Gly Thr Ile Val Ile Gly Gly Glu Met Asp
305 310 315 320
Glu Ala Thr Lys Phe Ile Ala Pro Thr Ile Val Lys Asp Val Pro Thr
325 330 335
Asn Asp Ser Leu Met Asn Glu Glu Ile Phe Gly Pro Val Leu Pro Ile
340 345 350
Val Pro Val Lys Asp Val Glu Glu Ala Ile Ala Tyr Val Asn Ser Asn
355 360 365
Asp His Pro Leu Ala Val Tyr Val Phe Ser Gln Asp Ala Ala Tyr Lys
370 375 380
Gln Lys Val Phe Ser Arg Thr Gln Ser Gly Ser Ala Val Ala Asn Glu
385 390 395 400
Val Val Ile Gln Pro Gly Ile Glu Gly Leu Pro Phe Gly Gly Ile Gly
405 410 415
Pro Ser Gly Ser Gly Tyr His Thr Gly Lys Tyr Thr Phe Asp Met Phe
420 425 430
Thr His Leu Arg Ala Ser Leu Asp Ser Pro Gly Trp Leu Asp Lys Ile
435 440 445
Leu Gly Phe Arg Phe Pro Pro Tyr Thr Asp Lys Ser Ile Lys Ala Ser
450 455 460
Gln Arg Ile Leu Lys Ser Leu Pro Pro Arg Pro Thr Gly Pro Pro Arg
465 470 475 480
Thr Asn Asn Ala Met Ala Asn Gly Ser Ala Thr Lys Trp Trp Gly Lys
485 490 495
Tyr Phe Phe Leu Ala Phe Val Leu Ala Thr Ile Gly Gly Leu Thr Lys
500 505 510
Pro Val Lys Ile Leu Gly Arg Lys Phe Val Pro Lys Ile Leu Gly
515 520 525
<210> 106
<211> 487
<212> PRT
<213> Phanerochaete carnosa
<400> 106
Met Thr Arg Leu Glu Tyr Thr Asn Ile Asp Asn Val Pro Lys Ile His
1 5 10 15
Ala Glu Leu Lys Glu Thr Phe Arg Ser Gly Lys Thr Arg Pro Val Pro
20 25 30
Phe Arg Lys Glu Gln Leu Ala Gln Leu Ala Trp Leu Leu Lys Asp Asn
35 40 45
Thr Asp Arg Ile Ala Asp Ala Ile His Thr Asp Leu Gly Arg Pro Ala
50 55 60
Ile Glu Ser Asp Leu Leu Asp Ile Asn Pro Ser Ile Gly Glu Ala Lys
65 70 75 80
Asp Ala Tyr Asp Asn Val Ala Lys Trp Ala Lys Thr Glu Lys Ala Arg
85 90 95
Trp Thr Phe Asn Phe Phe Ala Met Arg Pro Lys Ile Arg Lys Glu Pro
100 105 110
Lys Gly Val Val Leu Ile Ile Ser Pro Phe Asn Phe Pro Val Leu Leu
115 120 125
Leu Leu Gly His Leu Ala Ser Ala Leu Ala Ala Gly Asn Thr Val Val
130 135 140
Leu Lys Pro Ser Glu Leu Val Pro Ala Thr Ser Gln Leu Ile Ser Asp
145 150 155 160
Leu Ile Ser Gln Tyr Met Asp Pro Gly Val Val Arg Val Val Asn Gly
165 170 175
Asp Val Ser Val Thr Thr Lys Leu Leu Glu Leu Pro Trp Asp His Ile
180 185 190
Leu Tyr Thr Gly Ser Ser Arg Val Ala Lys Ile Ile Cys Thr Ala Ala
195 200 205
Ala Lys His Leu Thr Pro Val Thr Thr Glu Leu Gly Gly Lys Ser Pro
210 215 220
Val Ile Ile Asp Pro Lys Cys Asp Met Lys Leu Ala Ala Arg Arg Ile
225 230 235 240
Leu Trp Gly Lys Ile Ala Asn Ala Gly Gln Thr Cys Val Ala Pro Asp
245 250 255
Tyr Val Leu Val Pro Arg Glu Ala Gln Asp Ala Leu Val Asn Glu Leu
260 265 270
Met Glu Val Tyr Gln Thr Phe Tyr Pro Glu Gly Asp Pro Ala Thr Ser
275 280 285
Ala Ser Phe Ser Arg Ile Val Ser Gln Ala His Thr Thr Arg Ile Lys
290 295 300
Arg Leu Ile Asp Gly Thr Lys Gly Thr Val Val Val Gly Gly Thr Val
305 310 315 320
Asp Val Glu Lys Arg Tyr Ile Ala Pro Thr Ile Ile Arg Asp Val Pro
325 330 335
Val Asp Asp Ser Thr Met Asp Glu Glu Ile Phe Gly Pro Val Leu Pro
340 345 350
Ile Val Pro Val Arg Asp Val Glu Glu Ala Ile Gln Ile Val Asn Ser
355 360 365
Arg Asp His Pro Leu Ser Leu Tyr Val Phe Thr Gln Asp Ala Ala Phe
370 375 380
Lys Glu Asn Val Phe Ser Arg Thr Gln Ser Gly Ala Ala Leu Ala Asn
385 390 395 400
Glu Val Leu Val His Val Gly Ala Thr Gly Leu Pro Phe Gly Gly Ile
405 410 415
Gly Pro Ser Gly Ser Gly Ser Leu Thr Gly Lys His Gly Phe Asp Ala
420 425 430
Phe Thr His Leu Arg Ser Thr Leu Asp Asn Pro Lys Trp Val Asp Ala
435 440 445
Ile Met Lys Gly Arg Tyr Pro Pro Tyr Thr Pro Glu Lys Leu Ala Arg
450 455 460
Leu Arg Thr Ala Leu Lys Val Arg Met Pro Pro Arg Pro Gly Gly Ala
465 470 475 480
Arg Pro Ala Gln Lys Ser Ala
485
<210> 107
<211> 514
<212> PRT
<213> malassezia globosa
<400> 107
Met Lys Thr Thr Pro Ala Asp Thr Ile Pro Ala Leu Val Gly Arg Leu
1 5 10 15
Arg Ala Thr Phe Leu Thr Gly Lys Thr Arg Cys Val Glu Tyr Arg Lys
20 25 30
Asn Gln Leu Lys Gln Leu Tyr Phe Leu Val Lys Asp Asn Glu Glu Ala
35 40 45
Phe Val Asp Ala Ile Gly Gln Asp Leu Gly Arg Pro Gly Met Glu Thr
50 55 60
Thr Phe Ala Glu Val Ile Gly Ile Glu Asn Asp Leu Ala Thr Ser Ile
65 70 75 80
Ser Gln Leu Ser Lys Trp Ser Lys Asp Glu Cys Val Gly Ala Gly Pro
85 90 95
Pro Phe Met Leu His Gly Thr Lys Ile Arg Lys Asp Pro Lys Gly Thr
100 105 110
Val Leu Val Leu Gly Ala Trp Asn Tyr Pro Ile Thr Val Gln Leu Gly
115 120 125
Pro Met Val Gly Ala Ile Ala Ala Gly Asn Thr Val Ile Leu Lys Pro
130 135 140
Ser Glu Leu Ser Pro His Thr Ala Gln Leu Ile Ala Asp Leu Trp Ser
145 150 155 160
Lys Tyr Met Asp Thr Glu Thr Thr Ala Val Val Asn Gly Gly Ile Pro
165 170 175
Glu Ala Thr Ala Leu Leu Asp Gln Arg Phe Glu His Ile Phe Tyr Thr
180 185 190
Gly Asn Gly Arg Val Gly Arg Ile Val Ala Glu Lys Ala Ala Arg Trp
195 200 205
Leu Cys Pro Val Ser Leu Glu Leu Gly Gly Lys Ser Pro Val Ile Val
210 215 220
Asp Ala Ser Ala Asp Leu Lys Ile Ala Ala His Arg Thr Leu Trp Ala
225 230 235 240
Lys Ala Phe Asn Ala Gly Gln Thr Cys Val Ala Pro Asp Tyr Cys Leu
245 250 255
Val Asp Arg Arg Val Gln Asp Lys Phe Ala His Glu Leu Leu Gln Ala
260 265 270
Gln Arg Glu Phe Trp Pro Ser Arg Asp His Gln Glu Arg Asp Phe Gly
275 280 285
Arg Ile Val Ser Asp Asn His Trp Lys Arg Ile His Ser Leu Val Ser
290 295 300
Ser Ser Lys Ala Glu Leu Val Val Gly Gly Thr Ala Gly Ala Asp Gln
305 310 315 320
Ala Lys Arg Phe Ile Pro Leu Thr Ile Leu Lys Asn Val Asp Ala Ser
325 330 335
Asp Ser Val Met Thr Asp Glu Ile Phe Gly Pro Val Leu Pro Ile Val
340 345 350
Pro Phe Asp Thr Ile Arg Asp Ala Val Asp Phe Val Asn Glu Arg Asp
355 360 365
Gln Pro Leu Ala Leu Tyr Val Phe Thr Ser Cys Asn Glu Thr Arg Asp
370 375 380
Tyr Ile Leu Ala Tyr Thr Arg Ser Gly Gly Val Val Arg Gly Asp Cys
385 390 395 400
Leu Leu His Tyr Ala Ile Asp Ser Leu Pro Phe Gly Gly Thr Gly Pro
405 410 415
Ser Gly Tyr Gly Ser Tyr His Gly Lys Ala Gly Phe Asp Cys Phe Thr
420 425 430
His Glu Arg Ala Val Val Asp Ala Pro Ser Tyr Gly Met Leu Gly Lys
435 440 445
Leu Val Glu Val Val Met Ala Arg Arg Tyr Pro Pro Tyr Ser Lys Ser
450 455 460
Lys Leu Asp Phe Phe Arg Phe Val Leu Pro Lys Leu Val Trp Phe Gly
465 470 475 480
Arg Pro Pro Gln Pro Thr Arg Ser Ser Lys Ser Ile Asp His Pro Pro
485 490 495
Ser Lys Val Arg Thr Thr Ala Pro Arg His Gly Gly His Pro Leu Ser
500 505 510
Leu Val
<210> 108
<211> 538
<212> PRT
<213> Sporisorium reilianum
<400> 108
Met Ala Ala Ala Ala Thr Ala Ala Ala Glu Ala Gly Leu Gln Tyr Thr
1 5 10 15
Pro Ile Asp Asp Ile Pro Lys Ile Val Ser Asp Leu Arg Ala Ala Phe
20 25 30
Leu Thr Gly Lys Thr Arg Ser Ile Glu Tyr Arg Lys Asn Gln Leu Lys
35 40 45
Gln Leu Ala Tyr Met Leu Lys Asp His Gln Asp Asp Phe Ile Gln Ser
50 55 60
Leu Gln Lys Asp Leu Gly Arg Ser Arg Phe Glu Ser Ile Phe Ala Glu
65 70 75 80
Leu Met Gly Thr Thr Asn Glu Ile Val Glu Ala Val His Asn Leu Asp
85 90 95
Lys Trp Ala Lys Pro Ser Lys Pro Trp Ala Gly Ala Ala Trp Ala Met
100 105 110
His Gly Ala Ser Ile Arg Ser Glu Pro Lys Gly Thr Val Leu Val Leu
115 120 125
Gly Ala Trp Asn Tyr Pro Ile Thr Val Gln Ile Gly Pro Val Val Gly
130 135 140
Ala Ile Ala Ala Gly Asn Thr Val Val Leu Lys Pro Ser Glu Val Ala
145 150 155 160
Ser His Thr Ala Lys Leu Ile Ala Glu Leu Trp Asn Lys Tyr Leu Asp
165 170 175
Pro Glu Cys Tyr Arg Ile Ile Asn Gly Ala Ile Pro Glu Thr Thr Ala
180 185 190
Ala Leu Asp Gln Arg Tyr Glu His Ile Phe Tyr Thr Gly Asn Gly Thr
195 200 205
Val Gly Arg Ile Ile Ala Glu Lys Ala Ala Lys Trp Leu Cys Pro Thr
210 215 220
Thr Leu Glu Leu Gly Gly Lys Ser Pro Val Tyr Val Asp Lys Ser Ala
225 230 235 240
Asp Leu Lys Ile Ala Ala His Arg Ile Leu Trp Gly Lys Ser Phe Asn
245 250 255
Cys Gly Gln Thr Cys Ile Ala Pro Asp Tyr Val Leu Ile Pro His Glu
260 265 270
Leu Gln His Lys Phe Val His Glu Leu His Lys Ala Tyr Glu Arg Phe
275 280 285
Tyr Pro Glu Ile Lys Gly Gly Val Ser Gln Ser Glu Ser Tyr Ala Arg
290 295 300
Ile Ile Asn Pro Gly His Trp Lys Arg Leu Thr Ala Met Leu Ser Gly
305 310 315 320
Thr Lys Gly Lys Ile Val Leu Gly Gly Glu Gly Asp Glu Ala Thr Lys
325 330 335
Phe Leu Pro Pro Thr Val Ile Ala Asn Val Lys Thr Asp Asp Pro Val
340 345 350
Met Ala Gly Glu Ile Phe Gly Pro Leu Leu Pro Ile Val Pro Val Lys
355 360 365
Asp Val Lys Ala Ala Val Asp Phe Ile Asn Ser Arg Asp Gln Pro Leu
370 375 380
Ala Leu Tyr Leu Phe Ala Gly Asp Ser Lys Val Lys Glu Tyr Phe Phe
385 390 395 400
Asp Asn Thr Arg Ser Gly Ala Cys Val Gln Gly Asp Thr Leu Leu His
405 410 415
Phe Ala Val Asp Ala Leu Pro Phe Gly Gly Thr Gly Pro Ser Gly Tyr
420 425 430
Gly Asn Tyr His Gly Lys Ala Ser Phe Asp Gln Phe Ser His Gln Arg
435 440 445
Ala Ser Leu Asp Ala Pro Ser Thr Gly Leu Leu Gly Lys Ile Ile Glu
450 455 460
Ile Val Met Ser Ser Arg Tyr Pro Pro Tyr Thr Asn Ala Asn Leu Asn
465 470 475 480
Lys Met Arg Met Leu Ala Glu Tyr Ser Val Ser Phe Lys Arg Pro Ser
485 490 495
Asn Pro His Lys Ser Thr Thr Ser Ser Ser Ser Gly Gln Gly Ala Val
500 505 510
Ala Lys Arg Leu Ala Val Val Leu Leu Leu Ile Ser Leu Val Leu Gly
515 520 525
Ala Arg Asn Arg Gly Leu Ile Gly Trp Ile
530 535
<210> 109
<211> 640
<212> PRT
<213> Ustilago maydis
<400> 109
Met Arg Glu Trp Leu Gly Gly Asn Leu Arg Phe Ala Ala Ile Phe Ile
1 5 10 15
Arg Ile Gln Ser Arg Pro Asp His Ser Leu Arg Phe Thr Val Leu Asp
20 25 30
Pro Leu Pro Val His Tyr Arg Leu Ser Pro Pro Ser Ser Thr Leu Ala
35 40 45
Arg Leu His Gln Pro Arg Leu Ala Leu Thr Ser Phe Ala His Leu Ala
50 55 60
Leu Leu Pro Ser Ser Ile Ser Pro Ser Pro Thr Thr Leu Arg Leu Val
65 70 75 80
Val Cys Leu Val Thr Asn Ser His His Ser Val Ser Ser Ser Leu Lys
85 90 95
Arg Gln Ser Pro Ile Met Ala Ala Ala Ala Ala Thr Ala Ala Thr Glu
100 105 110
Ala Gly Leu Gln Tyr Thr Pro Ile Asp Asp Ile Pro Ser Ile Val Ser
115 120 125
Asp Leu Arg Ala Ala Phe Leu Thr Gly Lys Thr Arg Ser Val Glu Tyr
130 135 140
Arg Lys Asn Gln Leu Lys Gln Leu Ala Tyr Met Ile Lys Asp Asn Gln
145 150 155 160
Glu Ala Phe Val Glu Ser Leu Arg Lys Asp Leu Gly Arg Ser Arg Phe
165 170 175
Glu Ser Ile Phe Ala Glu Leu Met Gly Thr Thr Asn Glu Ile Val Glu
180 185 190
Ala Val Thr Lys Ile Asp Lys Trp Ala Lys Pro Ala Lys Pro Trp Ala
195 200 205
Gly Ala Ala Trp Ala Met His Gly Ala Thr Ile Arg Ser Glu Pro Lys
210 215 220
Gly Thr Val Leu Val Leu Gly Ala Trp Asn Tyr Pro Ile Thr Val Gln
225 230 235 240
Ile Gly Pro Val Ile Gly Ala Ile Ala Ala Gly Asn Thr Val Ile Leu
245 250 255
Lys Pro Ser Glu Val Ala Ser His Thr Ala Lys Leu Ile Ala Glu Leu
260 265 270
Trp Asn Lys Tyr Leu Asp Pro Glu Cys Phe Arg Val Val Asn Gly Gly
275 280 285
Ile Pro Glu Thr Thr Ala Leu Leu Asp Gln Arg Phe Glu His Ile Phe
290 295 300
Tyr Thr Gly Asn Gly Thr Val Gly Arg Ile Ile Ala Glu Lys Ala Ala
305 310 315 320
Lys Trp Leu Cys Pro Thr Thr Leu Glu Leu Gly Gly Lys Ser Pro Val
325 330 335
Tyr Val Asp Lys Ser Ala Asp Leu Ser Ile Ala Ala His Arg Ile Leu
340 345 350
Trp Gly Lys Ser Phe Asn Cys Gly Gln Thr Cys Ile Ala Pro Asp Tyr
355 360 365
Val Leu Ile Gln Pro Asp Leu Gln Asp Lys Phe Val Gln Glu Leu Lys
370 375 380
Lys Ala Tyr Gln Arg Phe Tyr Pro Glu Leu Gln Gly Gly Val Asn Asn
385 390 395 400
Ser Glu Ser Tyr Ala Arg Ile Ile Asn Pro Gly His Trp Lys Arg Leu
405 410 415
Asn Ala Met Leu Ser Gly Thr Lys Gly Lys Val Val Leu Gly Gly Glu
420 425 430
Gly Glu Glu Ala Thr Lys Phe Leu Pro Pro Thr Val Ile Ala Asp Val
435 440 445
Lys Pro Asp Asp Ala Ile Met Ser Gly Glu Ile Phe Gly Pro Leu Leu
450 455 460
Pro Ile Val Pro Val Arg Asp Val Glu Ala Ala Val Asp Leu Ile Asn
465 470 475 480
Ser Arg Asp Gln Pro Leu Ala Leu Tyr Leu Phe Ala Gly Asp Asn Arg
485 490 495
Val Lys Asn Phe Phe Phe Asp Asn Thr Arg Ser Gly Ala Cys Val Gln
500 505 510
Gly Asp Thr Leu Leu His Phe Ala Val Asp Val Leu Pro Phe Gly Gly
515 520 525
Thr Gly Pro Ser Gly Tyr Gly Asn Tyr His Gly Lys Ala Ser Phe Asp
530 535 540
Gln Phe Ser His Gln Arg Ala Ser Leu Asp Ala Pro Ser Thr Gly Leu
545 550 555 560
Leu Gly Lys Leu Val Glu Leu Ile Met Ser Ser Arg Tyr Pro Pro Tyr
565 570 575
Thr Glu Ala Asn Leu Lys Lys Leu Arg Ala Leu Ala Ala Tyr Ser Val
580 585 590
Ser Phe Lys Arg Pro Ser Asn Pro His Lys Ser Ile Ala Ser Ser Ser
595 600 605
Val Ser Leu Cys Leu Ser His Ser Arg Pro Ser Pro Phe Leu Ser Met
610 615 620
Ser Gln Ser Leu Phe Pro Met Val His Tyr Asn Met Leu Pro Thr Gln
625 630 635 640
<210> 110
<211> 578
<212> PRT
<213> Melampsora larici-populina
<400> 110
Met Ser Leu Ser Ser Lys Ala Gln Asp Gln Leu Asn Ser Gln Phe Thr
1 5 10 15
Ser Ile Asp Glu Ile Pro Lys Ile His Gln Glu Leu Arg Lys Ala Phe
20 25 30
Ser Asn Arg Ile Thr Lys Thr Leu Glu Trp Arg Thr His Gln Leu Lys
35 40 45
Gln Leu Gly Phe Leu Leu Gln Asp Asn Glu Gln Leu Ile Glu Glu Ala
50 55 60
Leu Ala Ile Asp Leu Gly Lys Pro Lys Thr Glu Ser His Ile Gly Glu
65 70 75 80
Leu Val Gly Thr Arg His Glu Val Leu Tyr Ala Leu Asn Asn Val Lys
85 90 95
Ser Trp Met Glu Pro Gln Ser Val Lys Thr Asp Leu Ala Trp Leu Val
100 105 110
Thr Lys Pro Lys Thr Phe His Glu Pro Lys Gly Val Val Leu Ile Phe
115 120 125
Gly Thr Trp Asn Tyr Pro Ile Ser Leu Ser Ile Ile Pro Leu Val Gly
130 135 140
Ala Ile Ala Gly Gly Asn Ala Val Val Leu Lys Leu Ser Glu Gln Ala
145 150 155 160
Pro Ala Ile Ala Asn Leu Leu Thr Lys Leu Ile Pro Gln Tyr Leu Asp
165 170 175
Asn Asn His Ile Arg Val Val Asn Gly Ala Ala Asp His Cys Asn Ala
180 185 190
Leu Leu Asp Leu Lys Phe Asp His Ile Phe Phe Thr Gly Ser Thr Gln
195 200 205
Val Gly Arg Thr Val Ala Lys Arg Ala Ala Glu His Met Thr Pro Val
210 215 220
Thr Leu Glu Leu Gly Gly Lys Ser Pro Ala Ile Val Phe Asp Asp Ala
225 230 235 240
Asp Phe Pro Val Ile Ala Arg Arg Leu Ile Trp Gly Lys Gly Met Asn
245 250 255
Ala Gly Gln Thr Cys Val Ala Pro Asp Tyr Ile Leu Val Ser Lys Lys
260 265 270
Ser Glu Ala Lys Leu Ile Thr Ser Leu Lys Lys Ala Met Gln Glu Leu
275 280 285
Tyr Pro Leu Asp Ser Ala Ser Gly Leu Ser Ala Lys Lys Ile Ile Asn
290 295 300
Ala Ser Gly Lys Ser Gly Asp Gly Pro Asp Asp Leu Gln Tyr Ser Lys
305 310 315 320
Ile Val Asn Gln Asn Gln Phe Asn Arg Leu Asn Asn Val Leu Gln Glu
325 330 335
Thr Lys Gly Glu Phe Ile Pro Thr Asp Asp Thr Phe Gln Ala Gly Thr
340 345 350
Lys Asp Ser Ser Asp Ala Gln Glu Leu Lys Met Pro Leu Thr Leu Val
355 360 365
Arg Asn Leu Thr Met Glu Asp Pro Val Met Gln Asn Glu Ile Phe Gly
370 375 380
Pro Ile Phe Pro Ile Leu Thr Tyr Asp Leu Gln Ser Glu Ser Met Ala
385 390 395 400
Glu Ile Leu Arg Pro Ile Ala Asp Ala Glu Pro Leu Ala Leu Tyr Val
405 410 415
Phe Thr Gln Ser Ser Gln Asn Phe Glu Leu Val Arg Gln His Thr Lys
420 425 430
Ser Gly Gln Ile Met Cys Asn Asp Leu Leu Ile Gln Phe Ala Ile Pro
435 440 445
Gly Leu Pro Phe Gly Gly Ile Gly Gln Ser Gly Ser Gly Asn Tyr His
450 455 460
Gly Tyr Tyr Ser Phe Leu Thr Phe Thr Tyr Glu Arg Ser Ser Ala Asn
465 470 475 480
Leu Pro Thr Trp Ala Asp Phe Leu Phe Asn Ala Arg Tyr Pro Pro Tyr
485 490 495
Thr Pro Phe Lys Leu Lys Leu Phe Ser Ala Ile Met Gly Pro Ala Arg
500 505 510
Ile Lys Gly Lys Ser Asn Pro Gly Leu Val Pro Lys Ser Ala Glu Val
515 520 525
Gly Lys Arg Ser Trp Leu Pro Thr Leu Thr Pro Leu Ser Phe Ser Thr
530 535 540
Leu Leu Leu Ala Gly Tyr Tyr Ala Leu Ser Arg Arg Tyr Gly Ser Asp
545 550 555 560
Tyr Leu Lys Ile Trp Met Thr Arg Phe Ile Gly Ala Ile Lys Gln Ser
565 570 575
Asn Arg
<210> 111
<211> 576
<212> PRT
<213> Puccinia graminis f. sp. tritici
<400> 111
Met Ser Ser Thr Ser Pro Leu Glu Phe Thr Pro Thr Glu Gln Ile Lys
1 5 10 15
Glu Ile Tyr Ala Thr Val Ser Gln Gly His Ser Ser Gly Ile Thr Lys
20 25 30
Ser Tyr Glu Trp Arg Glu His Gln Leu Lys Gln Leu Gly Tyr Leu Leu
35 40 45
Gln Glu Asn Glu Ser Leu Leu Glu Glu Ala Leu Thr Ile Asp Leu Gly
50 55 60
Arg Pro Asn Leu Glu Asn His Val Gly Glu Leu Val Gly Thr Arg Asn
65 70 75 80
Glu Val Leu Ser Ala Leu Lys Asn Leu Lys Lys Trp Ile Lys Pro Gln
85 90 95
Ser Val Lys Thr Glu Leu Thr Trp Leu Ile Ala Lys Pro Arg Val Ser
100 105 110
His Glu Pro Lys Gly Ile Val Ala Ile Phe Gly Ala Trp Asn Tyr Pro
115 120 125
Val Ala Val Leu Phe Gly Pro Leu Val Gly Ala Ile Ala Gly Gly Asn
130 135 140
Ser Ile Ile Leu Lys Pro Ser Glu Asn Cys Pro Ala Thr Ser Asn Leu
145 150 155 160
Met Thr Thr Leu Val Arg Lys Tyr Leu Asp Pro Arg Asn Ile Cys Val
165 170 175
Val Asn Gly Gly Gln Glu Gln Ser Thr Ala Leu Leu Asp Cys Arg Phe
180 185 190
Asp His Ile Phe Phe Thr Gly Gly Thr Ser Ile Gly Lys Ile Ile Ala
195 200 205
Leu Lys Ala Ala Glu Thr Leu Thr Thr Thr Thr Leu Glu Leu Gly Gly
210 215 220
Lys Ser Pro Val Val Val Leu Asp Asp Ala Asp Phe Leu Val Ala Ala
225 230 235 240
Arg Arg Ile Leu Trp Ala Lys Gly Leu Asn Ala Gly Gln Thr Cys Ile
245 250 255
Ala Pro Asp Tyr Val Leu Val Ser Glu Gln Ser Glu Ser Lys Leu Ile
260 265 270
Ala Ala Met Lys Gln Val Leu Lys Glu Phe Phe Pro Pro Asp Ala Gln
275 280 285
Gly Asp Lys Ala Ser Thr Asn Arg Asp Thr Thr Asp Pro Ser Asp Ser
290 295 300
Lys Phe Cys Lys Ile Ile Asn Gln Arg His Phe Asp Arg Leu Asn Ser
305 310 315 320
Tyr Leu Ser Gln Thr Lys Gly Glu Ile Val Lys Leu Asp Leu Asn Ser
325 330 335
Ser Ala Gln Pro Glu Ser Ala Asp Pro Ala Ser Leu Lys Ile Pro Leu
340 345 350
Thr Leu Ile Arg Asn Val Gln His Asp Asp Ile Leu Met Glu Asn Glu
355 360 365
Leu Phe Gly Pro Leu Leu Pro Ile Leu Thr Tyr Asn Asn Asp His Glu
370 375 380
Asp Ile Val Gln Cys Leu His Arg Ile Ser Gln Ser Ala Pro Leu Ala
385 390 395 400
Leu Tyr Ala Phe Gly Gln Ser Glu Glu Lys Leu Glu Phe Ile Arg Arg
405 410 415
Gln Thr Lys Ser Gly Gln Phe Val Cys Asn Asp Leu Leu Ile Gln Phe
420 425 430
Asn Ile Pro Gly Leu Pro Phe Gly Gly Val Gly Ala Ser Gly Leu Gly
435 440 445
Asn Tyr His Gly Tyr Tyr Ser Phe Leu Ala Phe Thr Tyr Glu Arg Pro
450 455 460
Met Val Asn Phe Pro Phe Trp Ala Asp Val Leu Leu Lys Ser Arg Tyr
465 470 475 480
Pro Pro Tyr Thr Ser Phe Lys Phe Lys Phe Met Gln Ala Val Leu Gly
485 490 495
Pro Ser Lys Leu Lys Gly Lys Ser Asn Pro Asn Pro Pro Ala Leu Thr
500 505 510
Asp Pro Leu Asp Phe Lys Arg Leu Leu Asp Pro Ser Ser Thr Gly Trp
515 520 525
Leu Ala Lys Ile Pro Val Lys Leu Ser Leu Met Ala Leu Leu Phe Ala
530 535 540
Phe Tyr Cys Ser Arg Arg Gln Asp Ser Leu Gly Gln Lys Gly Leu Phe
545 550 555 560
Asn Ser Phe Lys Lys Val Gln Asp Gln Val Lys Gln Phe Ile Ser Ser
565 570 575
<210> 112
<211> 28
<212> PRT
<213> Puccinia triticina
<400> 112
Met Ser Ser Thr Ser Pro Leu Glu Phe Thr Pro Thr Asp His Asp Lys
1 5 10 15
Glu Ile Tyr Ala Thr Val Ser Cys Gly His Ser Ser
20 25
<210> 113
<211> 18
<212> PRT
<213> Puccinia striiformis
<400> 113
Met Ala Ser Leu Glu Thr Pro Thr Asp Glu Ile Lys Gln Ile Tyr Ala
1 5 10 15
Thr Val
<210> 114
<211> 505
<212> PRT
<213> Microbotryum violaceum
<400> 114
Ser Ser Asp Tyr Thr Pro Val Asp Gln Ile Glu Pro Ala Tyr Asp Glu
1 5 10 15
Ile Val Lys Thr Phe Leu Thr Gly Lys Thr Arg Ser Ile Lys Trp Arg
20 25 30
Lys Gln Gln Leu Lys Gln Leu Gly Phe Leu Ile Gln Asp Asn Glu Ala
35 40 45
Ala Phe Ile Lys Ala Leu Arg Ser Asp Leu Ser Arg Pro Glu Phe Glu
50 55 60
Thr Thr Ile Ala Glu Leu Asn Pro Leu Lys Thr Asp Val Asn Glu Ala
65 70 75 80
Tyr Asp His Leu Asp Lys Trp Ala Lys Pro Val Lys Ala Lys Thr Thr
85 90 95
Ala Val Trp Ala Leu Ala Asn Ala Tyr Ile Tyr Asn Glu Pro Lys Gly
100 105 110
Ala Val Phe Ile Ile Gly Thr Trp Asn Tyr Pro Leu Val Leu Leu Leu
115 120 125
Ser Pro Leu Val Gly Ala Ile Ala Ala Gly Cys Thr Ala Leu Leu Lys
130 135 140
Pro Ala Glu Gln Ala Pro Ala Val Ala Lys Leu Val Gln Glu Leu Leu
145 150 155 160
Pro Lys Tyr Leu Asp Thr Ser Ala Tyr Lys Ile Ile Leu Gly Ala Val
165 170 175
Asp Gln Val Thr Arg Ala Leu Glu Leu Lys Phe Asp His Ile Phe Tyr
180 185 190
Thr Gly Ser Gly Gly Ile Gly Lys Ile Ile Ala Arg Ala Ala Ala Glu
195 200 205
His Leu Thr Pro Phe Thr Leu Glu Leu Gly Gly Lys Ser Pro Ala Val
210 215 220
Val Phe Asp Asp Ala Asn Ile Asp Ile Thr Ala Arg Arg Ile Met Trp
225 230 235 240
Gly Lys Phe Val Asn Ser Gly Gln Thr Cys Ile Ser Pro Asp Tyr Val
245 250 255
Leu Cys Thr Ala Asp Val Gln Asp Lys Leu Val Ala Ala Met Gln Lys
260 265 270
Val Tyr Lys Glu Phe Thr Thr Asp Ala Lys Gly Gln Glu Lys Ser Met
275 280 285
Val Asn Gly Glu Gly Tyr Ala Arg Ile Val Asn Met Asn His Phe Gly
290 295 300
Arg Ile Ser Thr Met Leu Asp Glu Thr Lys Gly Arg Val Val Val Gly
305 310 315 320
Gly Gly Arg Asn Lys Glu Thr Gly Lys Ile Glu Thr Thr Ile Ile Ala
325 330 335
Asp Val Gly Ala Asp Asp Pro Leu Met Lys Gly Glu Ile Phe Gly Pro
340 345 350
Val Met Pro Ile Val Val Lys Gln Thr Lys Glu Glu Met Val Glu Phe
355 360 365
Ile Gln Glu Arg Asp Asn Pro Leu Ala Leu Tyr Val Phe Thr Gln Ser
370 375 380
Thr Lys Asn Arg Asp Tyr Ile Phe Glu Arg Thr Arg Ser Gly Gly Phe
385 390 395 400
Val Gln Asn Asp Thr Ile Leu His Phe Thr Ile Pro Gly Leu Pro Phe
405 410 415
Gly Gly Ala Gly Ala Ser Gly Ile Gly Ala Tyr His Gly Lys Trp Ser
420 425 430
Phe Asp Thr Phe Ser His Gln Arg Ala Ser Ala His Ile Pro Thr Trp
435 440 445
Met Asp Val Ala Leu Asn Ser Arg Tyr Pro Pro Tyr Thr Pro Lys Lys
450 455 460
Leu Lys Met Met Leu Leu Ala Thr Lys Ala Val Ile Lys Arg Glu Ser
465 470 475 480
Lys Trp Ser Leu Lys Ser Leu Phe Gly Val Leu Ala Val Val Ala Ala
485 490 495
Ile Val Arg Tyr Arg Gln Ser Lys Leu
500 505
<210> 115
<211> 15
<212> PRT
<213> Rhodotorula glutinis ATCC 204091
<400> 115
Met Gln Asp Thr Pro Ile Asp Ser Ile Pro Cys Ala Thr Tyr Val
1 5 10 15
<210> 116
<211> 504
<212> PRT
<213> Rhodosporidium toruloides NP11
<400> 116
Met Ala Ala Met Gln Asp Thr Pro Ile Asp Ser Ile Pro Gln Ala Tyr
1 5 10 15
Asp Thr Val Thr Lys Ala Phe Leu Ser Gly Lys Thr Arg Pro Ile Ala
20 25 30
Trp Arg Lys Ala Gln Ile Lys Lys Leu Gly Phe Leu Val Gln Asp Asn
35 40 45
Glu Asp Ala Phe Val Arg Ala Leu Glu Gln Asp Phe Gly Arg Pro Ala
50 55 60
Phe Glu Thr Ile Thr Ala Glu Ile Asn Pro Val Lys Ala Glu Ile Asn
65 70 75 80
Glu Val Tyr Asp His Leu Glu Lys Trp Ala Lys Pro Arg Arg Val Lys
85 90 95
Thr Ser Ala Thr Trp Tyr Ala Thr Lys Pro Thr Val Tyr Ser Glu Pro
100 105 110
Lys Gly Val Thr Leu Val Ile Gly Thr Trp Asn Tyr Pro Ile Thr Leu
115 120 125
Leu Leu Val Pro Leu Leu Gly Ala Ile Ser Ala Gly Cys Thr Ala Leu
130 135 140
Val Lys Pro Ala Glu Gln Ala Pro His Val Ala Ala Leu Val Ala Asp
145 150 155 160
Leu Leu Pro Lys Tyr Leu Asp Pro Thr Ala Phe Ile Cys Ile Asn Gly
165 170 175
Ala Ile Pro Gln Ala Thr Ala Leu Leu Lys Leu Lys Phe Asp His Ile
180 185 190
Phe Tyr Thr Gly Ser Gly Thr Val Gly Lys Ile Val Ala Arg Ala Ala
195 200 205
Ala Glu His Leu Cys Pro Val Thr Leu Glu Leu Gly Gly Lys Ser Pro
210 215 220
Ala Val Val Leu Asp Asp Ala Asp Ile Glu Val Val Ala Arg Arg Ile
225 230 235 240
Val Trp Ala Lys Phe Thr Asn Ala Gly Gln Ile Cys Ile Ser Thr Asp
245 250 255
Tyr Val Leu Thr Thr Pro Gln Thr Glu Pro Lys Leu Leu Glu Ala Leu
260 265 270
Lys Arg Ala Leu Ala Ala Phe Ser Ala Asn Pro Ala Ala Ser Ser Ser
275 280 285
Ser Glu Lys Ser Ser Thr Ser Leu Val His Asn Pro Asn Tyr Ser Arg
290 295 300
Ile Ile Asn Gln Asn His Tyr Asn Arg Val Ser Lys Leu Leu Asp Ala
305 310 315 320
Thr Lys Gly Glu Val Val Val Gly Gly Gly Arg Asp Glu Lys Glu Arg
325 330 335
Lys Ile Glu Val Thr Ile Val Arg Gly Val Lys Pro Asp Asp Ser Leu
340 345 350
Met Ser Glu Glu Ile Phe Gly Pro Val Leu Pro Ile Met Thr Leu Pro
355 360 365
Thr Leu Asp Asp Met Val Lys Phe Ile Gln Ser Arg Asp Thr Pro Leu
370 375 380
Ala Leu Tyr Val Phe Thr Gln Ser Lys Lys Asn Arg Asp Phe Ile Phe
385 390 395 400
Glu Arg Thr Arg Ser Gly Gly Phe Val Gln Asn Asp Val Leu Val Gln
405 410 415
Phe Met Ile Pro Gly Leu Pro Phe Gly Gly Thr Gly Ala Ala Gly Tyr
420 425 430
Gly Asn Tyr His Gly Arg Arg Thr Phe Asp Thr Phe Ser His Glu Arg
435 440 445
Ala Ser Ala Asn Val Pro Thr Trp Met Asp Met Ile Met Ala Ser Arg
450 455 460
Tyr Pro Pro Tyr Thr Gln Lys Lys Leu Lys Met Leu Leu Phe Ala Thr
465 470 475 480
Lys Ala Val Ile Lys Lys Pro Ser Lys Phe Gly Ser Ile Ser Arg Leu
485 490 495
Leu Lys Lys Leu Thr Gly Gln Ala
500
<210> 117
<211> 16
<212> PRT
<213> Rhodosporidium toruloides ATCC 10657
<400> 117
Met Gln Asp Ile Pro Ile Asp Ser Asp Pro Gln Ala Tyr Asp Thr Val
1 5 10 15
<210> 118
<211> 21
<212> DNA
<213> Rhodosporidium toruloides
<400> 118
gcgagggatg gcagtaagac g 21
<210> 119
<211> 28
<212> DNA
<213> Rhodosporidium toruloides
<400> 119
aaaggatcca acttgctcgc ccagtacc 28
<210> 120
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 120
tttaagcttc acgtacagcc tgtggtagcc 30
<210> 121
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 121
tttaggcctg gaggagtcga gcgtgagagt 30
<210> 122
<211> 29
<212> DNA
<213> Rhodosporidium toruloides
<400> 122
tttccatggc tgcctcgtcg gcactcgag 29
<210> 123
<211> 29
<212> DNA
<213> Rhodosporidium toruloides
<400> 123
tttgatatcc attacgcctt gaccgtcag 29
<210> 124
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 124
aaagagctcg gtgactgcat gctccgttac 30
<210> 125
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 125
aaaggatcct gatggagtag ttgggcacga 30
<210> 126
<211> 31
<212> DNA
<213> Rhodosporidium toruloides
<400> 126
tttaagcttc ctcctccttg atctttcgcc g 31
<210> 127
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 127
tttaggcctg acctttgcgt cctcccttca 30
<210> 128
<211> 44
<212> DNA
<213> Rhodosporidium toruloides
<400> 128
tcagaacaac accagatcac tcacaatggc cgctaccctc cgcc 44
<210> 129
<211> 26
<212> DNA
<213> Rhodosporidium toruloides
<400> 129
tttcatatgg ccgctaccct ccgcca 26
<210> 130
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 130
tttgatatct ctagggcatc gtctagagtc 30
<210> 131
<211> 33
<212> DNA
<213> Rhodosporidium toruloides
<400> 131
aaagagctct attgttcgac tagactgcgc cac 33
<210> 132
<211> 32
<212> DNA
<213> Rhodosporidium toruloides
<400> 132
aaaggatcca aggaggatat tgtgcacgag ga 32
<210> 133
<211> 24
<212> DNA
<213> Rhodosporidium toruloides
<400> 133
cgactccaaa tcacccagtt cctc 24
<210> 134
<211> 28
<212> DNA
<213> Rhodosporidium toruloides
<400> 134
tttaggcctg accgactttg acgacgac 28
<210> 135
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 135
aaagagctcg cggcactgta cttcactacg 30
<210> 136
<211> 28
<212> DNA
<213> Rhodosporidium toruloides
<400> 136
aaaggatcca cgagacctat ccaaacgc 28
<210> 137
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 137
tttaagctta aggtcaagtc caaggccaac 30
<210> 138
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 138
tttaggcctg gctgctggag aaacgaaact 30
<210> 139
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 139
aaaaggcctc attccctcga ctcgacgcat 30
<210> 140
<211> 21
<212> DNA
<213> Rhodosporidium toruloides
<400> 140
agaggaggaa gttgtgcagc a 21
<210> 141
<211> 30
<212> DNA
<213> Rhodosporidium toruloides
<400> 141
tttggatccg ttgcggcgag tcctgtcatc 30
<210> 142
<211> 28
<212> DNA
<213> Rhodosporidium toruloides
<400> 142
tttgagctcc ggagcgagta agacgagg 28
<210> 143
<211> 28
<212> DNA
<213> Rhodosporidium toruloides
<400> 143
tttccatggc ctcgtacgcc gcccatcc 28
<210> 144
<211> 29
<212> DNA
<213> Rhodosporidium toruloides
<400> 144
tttgatatcg gaagggaccg cgctagttg 29
<210> 145
<211> 24
<212> DNA
<213> Rhodosporidium toruloides
<400> 145
tttccatggt cgcaccgtcc cccg 24
<210> 146
<211> 24
<212> DNA
<213> Rhodosporidium toruloides
<400> 146
tttccatggt cgcaccgtcc cccg 24

Claims (33)

1.一种真菌宿主细胞，其中在所述真菌宿主细胞中总脂肪酸包含水平为至少9％的升高水平的多不饱和脂肪酸(PUFA)，其中所述真菌宿主细胞的基因组已被修饰使得所述真菌宿主细胞与具有未修饰的基因组的真菌宿主细胞相比具有减少的天然醛脱氢酶(ALD1)酶活性，并且其中所述真菌宿主是红冬孢酵母属(Rhodosporidium)或红酵母属(Rhodotorula)的物种，并且其中所述天然ALD1的氨基酸序列由SEQ ID NO:3示出的序列组成。
2.权利要求1的真菌宿主细胞，其中所述PUFA为α-亚麻酸(ALA)或γ-亚麻酸(GLA)。
3.权利要求1的真菌宿主细胞，其中在所述真菌宿主细胞中总脂肪酸包含水平为至少24％的PUFA。
4.权利要求1的真菌宿主细胞，其中在所述真菌宿主细胞中总脂肪酸包含水平为至少49％的PUFA。
5.权利要求1的真菌宿主细胞，其中所述天然ALD1由选自以下的核酸编码：
(a)由SEQ ID NO:1中示出的核苷酸序列组成的核酸；
(b)由SEQ ID NO:2中示出的核苷酸序列组成的核酸；
(c)与(a)的核酸具有至少90％序列相同性的核酸；以及
(d)与(b)的核酸具有至少90％序列相同性的核酸。
6.权利要求1-5中任一项的真菌宿主细胞，其中所述真菌宿主细胞具有缺陷的编码所述ALD1的基因，导致所述Ald1酶活性丧失。
7.权利要求6的真菌宿主细胞，其中所述缺陷的基因是由T-DNA插入、同源重组或定点诱变引起的。
8.权利要求1-5中任一项的真菌宿主细胞，其中所述真菌宿主细胞的基因组已被修饰以下调所述天然ALD1基因的表达，导致所述Ald1酶活性丧失或减少。
9.权利要求8的真菌宿主细胞，其中所述表达是通过人工转录阻遏物分子下调的。
10.权利要求9的真菌宿主细胞，其中所述表达是通过RNAi分子下调的。
11.权利要求10的真菌宿主细胞，其中所述表达是通过siRNA分子、shRNA分子、miRNA分子下调的。
12.权利要求9的真菌宿主细胞，其中所述表达是通过反义分子下调的。
13.权利要求1-5中任一项的真菌宿主细胞，其中所述真菌宿主细胞的基因组已被进一步修饰以包括至少两个表达盒，其中每个表达盒包含可操作地连接至编码参与脂肪酸生物合成的蛋白的核酸的启动子，并且其中所述蛋白选自：
(a)酰基-CoAδ-12去饱和酶；
(b)硬脂酰-CoA-δ-9-去饱和酶；
(c)ω-3去饱和酶；
(d)脂肪酸延长酶；
(e)酰基-CoA羧化酶(ACC)；
(f)ATP:柠檬酸裂解酶(ACL)；
(g)二酰甘油酰基转移酶(DGA)；
(h)苹果酸酶(MAE)；以及
(i)酰基-CoA δ-6去饱和酶。
14.权利要求13的真菌宿主细胞，其中所述表达盒进一步包含可操作地连接至编码参与脂肪酸生物合成的蛋白的核酸的转录终止子。
15.权利要求13的真菌宿主细胞，其中所述核酸的编码序列包含60％-70％C和G，并且其中至少70％的密码子在第3位具有C或G。
16.权利要求13的真菌宿主细胞，其中所述蛋白选自：
(a)酰基-CoA δ-12去饱和酶，其由SEQ ID NO:5或SEQ ID NO:94中示出的氨基酸序列组成；
(b)硬脂酰-CoA-δ-9去饱和酶，其由SEQ ID NO:8中示出的氨基酸序列组成；
(c)ω-3去饱和酶，其由SEQ ID NO:10或SEQ ID NO:12中示出的氨基酸序列组成；
(d)脂肪酸延长酶，其由SEQ ID NO:101或SEQ ID NO:104(Elo2)中示出的氨基酸序列组成；
(e)酰基-CoA羧化酶(ACC1)，其由SEQ ID NO:91中示出的氨基酸序列组成；
(f)ATP:柠檬酸裂解酶(ACL1)，其由SEQ ID NO:88中示出的氨基酸序列组成；
(g)二酰甘油酰基转移酶(DGA1)，其由SEQ ID NO:82中示出的氨基酸序列组成；
(h)苹果酸酶(MAE1)，其由SEQ ID NO:85中示出的氨基酸序列组成；以及
(i)酰基-CoA δ-16去饱和酶，其由SEQ ID NO:96或SEQ ID NO:98中示出的氨基酸序列组成。
17.权利要求16的真菌宿主细胞，其中所述蛋白由选自以下的核酸编码，所述核酸由SEQ ID NO:4、6、7、9、11、80、81、83、84、86、87、89、90、92、93、95、97、99、100、102和103中示出的核苷酸序列组成。
18.权利要求13的真菌宿主细胞，其中所述启动子是分离自编码选自以下蛋白的基因的启动子：甘油醛3-磷酸脱氢酶(GPD)、酰基-CoA载体蛋白(ACP)、脂肪酸去饱和酶、翻译延伸因子(TEF)、丙酮酸脱羧酶(PDC)、烯醇化酶(2-磷酸甘油酸脱水酶)(ENO)、肽基脯氨酰异构酶(PPI)、乙酰基-CoA羧化酶(ACC)和转醛醇酶。
19.权利要求18的真菌宿主细胞，其中所述启动子是选自SEQ ID NO:55-79中示出的启动子组的启动子序列。
20.权利要求18的真菌宿主细胞，其中所述启动子分离自红冬孢酵母属或红酵母属的物种。
21.权利要求1的真菌宿主细胞，其中由SEQ ID NO:99中示出的序列组成的ELO1基因或由SEQ ID NO:102中示出的序列组成的ELO2基因已被人工操纵为具有升高或降低水平的脂肪酸延长酶活性。
22.一种产生ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使权利要求1-5中任一项的真菌宿主细胞在适合产生PUFA的条件下生长。
23.一种产生三酰甘油(TAG)的方法，所述方法包括使权利要求1-5中任一项的真菌宿主细胞在适合产生TAG的条件下生长。
24.一种产生ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使权利要求6的真菌宿主细胞在适合产生PUFA的条件下生长。
25.一种产生ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使权利要求8的真菌宿主细胞在适合产生PUFA的条件下生长。
26.一种产生ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使权利要求13的真菌宿主细胞在适合产生PUFA的条件下生长。
27.一种产生ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使权利要求16的真菌宿主细胞在适合产生PUFA的条件下生长。
28.一种产生ω-3和ω-6多不饱和脂肪酸(PUFA)的方法，所述方法包括使权利要求21的真菌宿主细胞在适合产生PUFA的条件下生长。
29.一种产生三酰甘油(TAG)的方法，所述方法包括使权利要求6的真菌宿主细胞在适合产生TAG的条件下生长。
30.一种产生三酰甘油(TAG)的方法，所述方法包括使权利要求8的真菌宿主细胞在适合产生TAG的条件下生长。
31.一种产生三酰甘油(TAG)的方法，所述方法包括使权利要求13的真菌宿主细胞在适合产生TAG的条件下生长。
32.一种产生三酰甘油(TAG)的方法，所述方法包括使权利要求16的真菌宿主细胞在适合产生TAG的条件下生长。
33.一种产生三酰甘油(TAG)的方法，所述方法包括使权利要求21的真菌宿主细胞在适合产生TAG的条件下生长。
CN201580060087.4A 2014-09-08 2015-08-21 在红冬孢酵母属和红酵母属物种中高效产生多不饱和脂肪酸(pufa)的方法 Active CN107075450B (zh)

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PCT/SG2015/050273 WO2016039685A1 (en) 2014-09-08 2015-08-21 Methods for efficient production of polyunsaturated fatty acids (pufa) in rhodosporidium and rhodotorula species

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