脱氧-D-木酮糖磷酸生物合成途径的基因在改变类异戊二烯浓度中 的应用 本发明涉及来自于细菌或寄生虫的DNA序列(SEQ：1，3，5，7)的应用，该基因编码gcpE或yfgB蛋白质，当将该基因掺入到病毒，真核细胞和原核细胞的基因组时所述的DNA序列改变类异戊二烯的含量，并且涉及测量类异戊二烯合成中gcpE基因的活性的方法。此外本发明还涉及鉴定在植物中具有除草剂，抗寄生虫，抗病毒，杀真菌的作用和在人和动物中具有抗真菌，抗寄生虫，抗病毒的作用的物质的方法。
已经知道借助于经典的乙酸/甲羟戊酸途径和一种可替代的甲羟戊酸-依赖性生物合成途径，脱氧-D-木酮糖磷酸途径形成类异戊二烯的生物合成途径(Rohmer，M.，Knani，M.，Simonin，p.，Sutter，B.和Sahm，H.(1993)：生物化学杂志295：517-524)。
在美国专利US5858367中描述了aarC-寡聚核苷酸在识别抗细菌物质中的应用。
令人惊奇的是，已经发现gcpE蛋白质在类异戊二烯生物合成地另一种代谢途径中还具有激酶功能和催化类异戊二烯生物合成的糖或前体的磷酸化作用，特别是2-C-甲基-D-赤藓糖醇，2-C-甲基-D-赤藓糖醇磷酸的磷酸化，特别是2-C-甲基-D-赤藓糖醇-4-磷酸的磷酸化，2-C-甲基-D-赤藓糖，2-C-甲基-D-赤藓糖磷酸的磷酸化，特别是2-C-甲基-D-赤藓糖-4-磷酸，
CH2(OH)-C(CH3)＝C(OH)-CH2-O-PO(OH)2，
CH2(OH)-C(CH3)＝C(OH)-CH2-OH，
CH2(OH)-C(CH3)-CO-CH2-O-PO(OH)2，
CH2(OH)-C(CH3)-CO-CH2OH，CH2＝C(CH3)-CO-CH2-O-PO(OH)2，
CH2＝C(CH3)-CO-CH2-OH，CH2＝C(CH3)-CH(OH)-CH2-O-PO(OH)2，
CH2＝C(CH3)-CH(OH)-CH2-OH，
CH2(OH)-C(＝CH2)-C(OH)-CH2-O-PO(OH)2，
CH2(OH)-C(＝CH2)-C(OH)-CH2-OH，
CHO-CH(CH3)-CH(OH)-CH2-O-PO(OH)2，
CHO-CH(CH3)-CH(OH)-CH2-OH，
CH2(OH)-C(OH)(CH3)-CH＝CH-O-PO(OH)2，
CH2(OH)-C(OH)CH3-CH＝CH-OH，
CH(OH)＝C(CH3)-CH(OH)-CH2-O-PO(OH)2，
CH(OH)＝C(CH3)-CH(OH)-CH2-OH，
CH3-C(CH3)＝CH-CH2-O-PO(OH)2，CH3-C(CH3)＝CH-CH2-OH，
CH2＝C(CH3)-CH2-CH2-O-PO(OH)2，CH2＝C(CH3)-CH2-CH2-OH的磷酸化。
因此本发明涉及来自于细菌或寄生虫的DNA序列的应用，所述DNA序列编码gcpE或yfgB的蛋白质[来自于细菌或来自寄生虫的gcpE或yfgB]或所述的DNA序列编码该蛋白质的类似物或衍生物，其中该蛋白质的一个或多个氨基酸被缺失，叠加或被其它氨基酸替代，并且基本上没有降低该多肽的酶促活性。特别是本发明涉及SEQ1，3，5，7的DNA序列的应用。列举的SEQ 1和5序列以及蛋白质2和6的起始来源是微生物大肠杆菌K12菌株。SEQ 3和7列举的序列以及蛋白质4和8的原始来源是微生物恶性疟原虫株3D7。
这样的DNA序列描述于US5858367并且也可以通过下面的国际互联网地址和登记号找到http：//www3.ncbi.nlm.nih.gov/Entrez/protein.html AAD07695，AAD18517，AAC75568，AAC67648，AAC65433，P36979，CAA15530，CAA98356，CAA98355，AAC24056，AAC07467，P54482，P44667，P27434，P27433，BAA17717，BAA20919，BAA16402，S23058，AAB51469，1819264A，CAA45783，CAA45782，AAA21360，AAA21359，BAA02549，I39486，2113330A。
本发明的序列适用于在病毒、真核生物和原核生物中表达基因，他们负责1-脱氧-D-木酮糖途径的类异戊二烯的合成。
根据本发明真核生物或真核生物细胞包括动物细胞，植物细胞，藻类，酵母，真菌，而原核生物或原核生物细胞包括细菌，古细菌和真细菌。
当将DNA序列插入到定位了上面所述DNA序列的基因组时，上面所说的基因能够在病毒、真核生物和原核生物中表达。以本领域内技术人员已知的方式培养本发明转化的病毒、真核生物和原核生物并且分离在这样的培养期间形成的类异戊二烯，非强制性地进行纯化。不是所有的类异戊二烯都需要分离，在一些情况下，类异戊二烯可以直接释放到室温。
借助于下面的步骤可以进行所使用的转基因病毒、真核生物和原核生物的制备以便修饰类异戊二烯的含量：
a)生产具有下面亚序列的DNA序列
i)在病毒、真核生物和原核生物中具有活性并且在预定的靶组织或靶细胞中确保形成RNA，
ii)编码具有来自于细菌或寄生虫的gcpe或yfgB蛋白质的氨基酸序列的多肽或者该多肽的类似物或者衍生物的DNA序列，
iii)导致将聚-A残基加到病毒、真核生物和原核生物的RNA的3’末端的未翻译的序列，
b)利用或不利用载体(例如质粒，病毒DNA)将该DNA序列转移到和掺入到病毒细胞，原核生物细胞或真核生物细胞的基因组。
从这种方式转化的植物细胞再生完整的整个植株。
给编码gcpE或yfgB蛋白质或者其类似物或者衍生物的序列提供确保在一定的器官或细胞中转录的启动子，该启动子以有义方向偶合(启动子的3’末端到编码序列的5’末端)到编码待形成的蛋白质的序列。将决定mRNA合成的终止的终止信号结合到编码序列的3’末端。以便指导将表达的蛋白质到一定的亚细胞室，例如叶绿体，淀粉体，线粒体，液泡，胞液或者细胞间隔，将编码所谓的信号序列或转移肽的其他的序列插入到启动子和该编码序列之间。该序列必须是在与该蛋白质的编码序列相同的阅读框架中。为了将本发明的DNA序列导入到高等植物中需要获得大量的克隆载体，所述的载体含有大肠杆菌中的复制信号和用以选择转化细胞的标记物。载体的例子是pBR322，pUC-系列，M13mp-系列，pACYC184，EMBL3等等。根据将需要的基因导入到植物中的方法，需要其他的DNA序列。例如如果将Ti或Ri质粒用于转化植物细胞，必须插入Ti和Ri质粒T-DNA的至少一个右边界，但是通常是右边界和左边界，作为待导入基因的侧面区域。在欧洲专利120516；Hoekama在“双性植物载体系统”，Offset-drukkerij KantersB.V.Alblasserdam(1985)，第5章；Fraley等人，植物科学的关键和综述4，1-46和An等人(1985)EMBO J.4，277-287广泛地调查和描述的T-DNA在转化植物细胞中的应用。一旦将导入的DNA掺入到基因组，通常是稳定的并且还保留在原始转化的细胞的子代中。通常它含有选择性标记，该标记授予转化植物细胞对杀生剂或抗生素，例如卡那霉素，G418，bleomycin，潮霉素或磷苏菌素和其他的抗性。所使用的特定的标记物允许选择出失去插入的DNA的转化细胞。
许多技术可用于将DNA导入到植物。这些技术包括用农杆菌的辅助转化，原生质体的融合，DNA的微注射，电击穿，以及微弹轰击方法和病毒注射。然后将转化植物材料在合适的培养基上再生为完整植株，所述的培养基含有用于选择目的的抗生素或杀生剂。对于用于注射和电击穿的质粒没有特别的需求。但是，如果完整植株从这样的转化细胞再生，必须存在选择性标记物。以常规的方式在植物中生长转化细胞(McCormick等人(1986)，植物细胞报告5，81-84)。通常可以培养该植物并且与具有相同的转化基因组或其他基因组的植物进行杂交。获得的个体具有相应的表现型特性。
本发明还提供了表达载体，它含有一个或多个本发明的DNA序列。可以通过给本发明的DNA提供合适的功能调节信号获得这样的表达载体。这样的复制信号是负责表达的DNA序列，例如启动子，加强子，核糖体结合位点，并且由宿主生物体识别。
非强制性地其他的调节信号例如控制重组DNA在宿主生物体中复制或重组的，也可以是表达载体的组成部份。
用于表达本发明的酶的合适的宿主细胞和生物体是那些不包括具有DOXP合成酶功能的固有酶，DOXP还原异构酶或gcpE激酶的微生物。这样的例子是古细菌，动物，真菌，丝状毛霉和一些真细菌。这样的固有酶活性的缺失本质上促进了重组酶的检测和纯化。因此，也可能是利用各种化学品和药物第一次直接从宿主细胞的粗提物中检测到本发明的重组酶的活性和特别是活性的抑制。
如果想要获得多肽链的翻译后修饰和天然折叠本发明的酶优选地在真核细胞中表达。但是根据表达系统，当表达通过DNA拼接去除内含子的基因组DNA序列时，确保产生的酶的多肽序列对寄生虫具抗性。利用重组DNA技术，将编码内含子的序列去除或为了试验的目的将该序列插入到待表达的DNA序列。
利用本领域内技术人员已知的方法从宿主细胞或宿主细胞的上清液可以分离蛋白质。也可以需要酶的体外重新活化。
为了有助于纯化，以具有不同肽链的融合蛋白的形式表达本发明的酶活该酶的亚序列。寡聚一组氨酸序列和来源于谷胱甘肽S-转移酶，硫氧还蛋白或钙调素结合肽的序列特别适用于该目的。
进一步以具有本领域内技术人员已知的这样的肽链的融合蛋白的形式表达本发明的酶或该酶的亚序列，该重组酶被运输到细胞外或运输到宿主细胞的腔室。因此有助于该酶的思维活性的纯化和调查。
当表达本发明的酶时，证明修饰个体密码子是方便的。如果在寄生虫中使用的密码子不同于在异源表达系统中的密码子的使用，该编码序列中碱基的有目的替代是可取的，以便确保蛋白质的最佳合成。此外非翻译5’和3’区域的分别缺失经常可调节，例如如果存在DNA的3’序列的几个脱稳定的序列基序ATTTA。然后在真核生物中优选表达中应该缺失这些。碱基的缺失，叠加或替代时这类变异并且是本发明的主题。
进一步可以在标准条件下，采用本领域内技术人员已知的方法进行体外翻译获得本发明的酶。适用于本发明的目的的系统是兔子网织红血球和小麦胚提取物和细菌裂解液。也可以将体外转录的mRNA翻译到Xenopus卵子。
其序列来源于本发明的酶的肽序列的寡聚-和多肽的序列可以通过化学合成方法获得。如果合适选择该序列，这样的肽具有本发明特征的特性。这样的肽可以大量地产生并且特别适合于酶活性动力学，酶活性的调节，酶的三维空间结构，各种化学剂和药物对酶活性的抑制，各种配体的结合几何形状和结合亲和性的调查。
本发明也提供了用于测定gcpE蛋白质的酶促活性的方法。利用已知的方法可以测定所述的活性。通过测定糖或磷糖类或类异戊二烯生物合成的前体的磷酸化，特别是2-C-甲基-D-赤藓糖醇，2-C-甲基-D-赤藓糖醇磷酸，特别是2-C-甲基-D-赤藓糖醇4-磷酸的磷酸化，2-C-甲基-D-赤藓糖，2-C-甲基-D-赤藓糖磷酸的磷酸化，特别是2-C-甲基-D-赤藓糖4-磷酸，CH2(OH)-C(CH3)＝C(OH)-CH2-O-PO(OH)2，
CH2(OH)-C(CH3)＝C(OH)-CH2-OH，
CH2(OH)-C(CH3)-CO-CH2-O-PO(OH)2，
CH2(OH)-C(CH3)-CO-CH2OH，CH2＝C(CH3)-CO-CH2-O-PO(OH)2，
CH2＝C(CH3)-CO-CH2-OH，CH2＝C(CH3)-CH(OH)-CH2-O-PO(OH)2，
CH2＝C(CH3)-CH(OH)-CH2-OH，
CH2(OH)-C(＝CH2)-C(OH)-CH2-O-PO(OH)2，
CH2(OH)-C(＝CH2)-C(OH)-CH2-OH，
CHO-CH(CH3)-CH(OH)-CH2-O-PO(OH)2，
CHO-CH(CH3)-CH(OH)-CH2-OH，
CH2(OH)-C(OH)(CH3)-CH＝CH-O-PO(OH)2，
CH2(OH)-C(OH)CH3-CH＝CH-OH，
CH(OH)＝C(CH3)-CH(OH)-CH2-O-PO(OH)2，
CH(OH)＝C(CH3)-CH(OH)-CH2-OH，
CH3-C(CH3)＝CH-CH2-O-PO(OH)2，CH3-C(CH3)＝CH-CH2-OH，
CH2＝C(CH3)-CH2-CH2-O-PO(OH)2，CH2＝C(CH3)-CH2-CH2-OH的磷酸化进行测定。本发明还提供了将该测定方法用于识别抑制特定的酶的活性的物质。
已经发现脱氧-1-木酮糖磷酸代谢途径也存在于许多寄生虫，病毒和真菌。
因此本发明还涉及用于筛选抑制脱氧-D-木酮糖磷酸代谢途径的化合物的方法。根据本发明，提供了含有重组表达载体的宿主生物体，其中载体包括至少SEQ ID NO：1，SEQ ID NO：3或SEQID NO：5的寡聚核苷酸序列的一部分或其变异体或其同系物，和提供了被怀疑对人和动物具有抗微生物，抗寄生虫，抗细菌，抗病毒和抗真菌的作用或对植物具有抗微生物，抗病毒，杀菌剂，除草剂或杀真菌剂的作用的化合物。然后将宿主生物体与该化合物和待测定的化合物接触。实施例1借助于gcpE和yfgB的过量表达在大肠杆菌中积累的类胡萝卜素增加
首先根据公开出版的方案(Cunningham，FXjr等人，1996，植物细胞8：1613-1626)已经构建了质粒pAC-LYC。该质粒支持该基因，用于从IPP和DMAPP合成类胡萝卜素Lycopen是必要的。因此，已经用pAC-LYC转化大肠杆菌形成玫瑰红色的菌落。如果用于合成类胡萝卜素的离析物的可获得性增加，类胡萝卜素的积累增加，菌落看起来深红色。借助于DOXP途径的基因的过量合成可以形成离析物的加强形成。因此大肠杆菌的gcpE和yfgB基因在合适的表达载体中克隆。用引物5’-CCA TGG GCC ATA ACCAGG CTC CAA TCC AA-3’和5’-GGA TCC TTT TTC AAC CTGCTG AAC GTC AAT-3’进行PCR从大肠杆菌的基因组DNA扩增gcpE基因并且克隆到pCR2.1-TOPO载体。借助于限制性界面NcoI和BamHI将插入物克隆到表达载体pQE60。用引物5’-GGA TCCATG TCT GAA CAA TTA GTC ACA-3’和5’-AAG CTT TCA GACCGC TTT AAT GTC GAT GGC-3’将yfgB基因进行扩增并且克隆到pCRT7/NT TOPO载体。借助于限制性界面BamHI和HindIII将插入物克隆到表达载体pQE30。已经用pAC-LYC和两个表达载体之一转化的细菌显示比用仅仅用空pQE30转化的对照有显著的较深的着色。对类胡萝卜素的丰富度进行光密度计测定与对照相比产生210％的gcpe和173％的yfgB。实施例2
借助于gcpe和yfgB基因的过量表达在恶性疟原虫中积累的类胡萝卜素增加
通过利用恶性疟原虫的gcpe和yfgB基因已经进行了类似的试验。用引物5’-CTG ATT CTT TTT ATG TTA CTG TTT TAT TCTCAT GTA-3’和5’-CTA CCC TTT TTT ATT TGT AAG AAC ATCATT AGT TAC GTT AAC-3’已经扩增了gcpe基因并且用引物GAAAAG TCA AAA AGG TAC ATA AGC CTG ATT AAG和5’-TAA CATGTC CTC TAT GCC TAT ATA TTT ATA TAA TG-3’已经扩增了yfgB基因。恶性疟原虫菌株3D7的基因组DNA已经用作为模板并且已经将热稳定的Pwo-DNA聚合酶用于PCR。通过T4-聚核苷酸激酶将PCR产物磷酸化并且克隆到已经用SmaI线性化的pQE32-载体，并且由碱性磷酸酶进行脱磷酸化。通过限制性分析证实插入物的定向。用这些构建物获得的细菌菌落没有显示颜色的变化。但是，光密度分析产生了117％(gcpe)和113％(yfgB)的类胡萝卜素的积累。与恶性疟原虫基因相比类胡萝卜素的较少积累明显是由于经常观察到由于高A/T-含量在大肠杆菌中恶性疟原虫基因的较少表达。
序列表<110>哈桑·朱马<120>脱氧-D-木酮糖磷酸生物合成途径的基因在改变异戊二烯类的浓度中的应用<130>15904<140><141><150>19923567.8<151>1999-05-21<150>19923568.6<151>1999-05-21<160>8<170>PatentIn Ver.2.1<210>1<211>1119<212>DNA<213>大肠杆菌<220><221>CDS<222>(1)..(1119)<400>1atg cat aac cag gct cca att caa cgt aga aaa tca aca cgt att tac 48Met His Asn Gln Ala Pro Ile Gln Arg Arg Lys Ser Thr Arg Ile Tyr 1 5 10 15gtt ggg aat gtg ccg att ggc gat ggt gct ccc atc gcc gta cag tcc 96Val Gly Asn Val Pro Ile Gly Asp Gly Ala Pro Ile Ala Val Gln Ser
20 25 30atg acc aat acg cgt acg aca gac gtc gaa gca acg gtc aat caa atc 144Met Thr Asn Thr Arg Thr Thr Asp Val Glu Ala Thr Val Asn Gln Ile
35 40 45aag gcg ctg gaa cgc gtt ggc gct gat atc gtc cgt gta tcc gta ccg 192Lys Ala Leu Glu Arg Val Gly Ala Asp Ile Val Arg Val Ser Val Pro
50 55 60acg atg gac gcg gca gaa gcg ttc aaa ctc atc aaa cag cag gtt aac 240Thr Met Asp Ala Ala Glu Ala Phe Lys Leu Ile Lys Gln Gln Val Asn 65 70 75 80gtg ccg ctg gtg gct gac atc cac ttc gac tat cgc att gcg ctg aaa 288Val Pro Leu Val Ala Asp Ile His Phe Asp Tyr Arg Ile Ala Leu Lys
85 90 95gta gcg gaa tac ggc gtc gat tgt ctg cgt att aac cct ggc aat atc 336Val Ala Glu Tyr Gly Val Asp Cys Leu Arg Ile Asn Pro Gly Asn Ile
100 105 110ggt aat gaa gag cgt att cgc atg gtg gtt gac tgt gcg cgc gat aaa 384Gly Asn Glu Glu Arg Ile Arg Met Val Val Asp Cys Ala Arg Asp Lys
115 120 125aac att ccg atc cgt att ggc gtt aac gcc gga tcg ctg gaa aaa gat 432Asn Ile Pro Ile Arg Ile Gly Val Asn Ala Gly Ser Leu Glu Lys Asp
130 135 140ctg caa gaa aag tat ggc gaa ccg acg ccg cag gcg ttg ctg gaa tct 480Leu Gln Glu Lys Tyr Gly Glu Pro Thr Pro Gln Ala Leu Leu Glu Ser145 150 155 160gcc atg cgt cat gtt gat cat ctc gat cgc ctg aac ttc gat cag ttc 528Ala Met Arg His Val Asp His Leu Asp Arg Leu Asn Phe Asp Gln Phe
165 170 175aaa gtc agc gtg aaa gcg tct gac gtc ttc ctc gct gtt gag tct tat 576Lys Val Ser Val Lys Ala Ser Asp Val Phe Leu Ala Val Glu Ser Tyr
180 185 190cgt ttg ctg gca aaa cag atc gat cag ccg ttg cat ctg ggg atc acc 624Arg Leu Leu Ala Lys Gln Ile Asp Gln Pro Leu His Leu Gly Ile Thr
195 200 205gaa gcc ggt ggt gcg cgc agc ggg gca gta aaa tcc gcc att ggt tta 672Glu Ala Gly Gly Ala Arg Ser Gly Ala Val Lys Ser Ala Ile Gly Leu
210 215 220ggt ctg ctg ctg tct gaa ggc atc ggc gac acg ctg cgc gta tcg ctg 720Gly Leu Leu Leu Ser Glu Gly Ile Gly Asp Thr Leu Arg Val Ser Leu225 230 235 240gcg gcc gat ccg gtc gaa gag atc aaa gtc ggt ttc gat att ttg aaa 768Ala Ala Asp Pro Val Glu Glu Ile Lys Val Gly Phe Asp Ile Leu Lys
245 250 255tcg ctg cgt atc cgt tcg cga ggg atc aac ttc atc gcc tgc ccg acc 816Ser Leu Arg Ile Arg Ser Arg Gly Ile Asn Phe Ile Ala Cys Pro Thr
260 265 270tgt tcg cgt cag gaa ttt gat gtt atc ggt acg gtt aac gcg ctg gag 864Cys Ser Arg Gln Glu Phe Asp Val Ile Gly Thr Val Asn Ala Leu Glu
275 280 285caa cgc ctg gaa gat atc atc act ccg atg gac gtt tcg att atc ggc 912Gln Arg Leu Glu Asp Ile Ile Thr Pro Met Asp Val Ser Ile Ile Gly
290 295 300tgc gtg gtg aat ggc cca ggt gag gcg ctg gtt tct aca ctc ggc gtc 960Cys Val Val Asn Gly Pro Gly Glu Ala Leu Val Ser Thr Leu Gly Val305 310 315 320acc ggc ggc aac aag aaa agc ggc ctc tat gaa gat ggc gtg cgc aaa 1008Thr Gly Gly Asn Lys Lys Ser Gly Leu Tyr Glu Asp Gly Val Arg Lys
325 330 335gac cgt ctg gac aac aac gat atg atc gac cag ctg gaa gca cgc att 1056Asp Arg Leu Asp Asn Asn Asp Met Ile Asp Gln Leu Glu Ala Arg Ile
340 345 350cgt gcg aaa gcc agt cag ctg gac gaa gcg cgt cga att gac gtt cag 1104Arg Ala Lys Ala Ser Gln Leu Asp Glu Ala Arg Arg Ile Asp Val Gln
355 360 365cag gtt gaa aaa taa 1119Gln Val Glu Lys
370<210>2<211>372<212>PRT<213>大肠杆菌<400>2Met His Asn Gln Ala Pro Ile Gln Arg Arg Lys Ser Thr Arg Ile Tyr 1 5 10 15Val Gly Asn Val Pro Ile Gly Asp Gly Ala Pro Ile Ala Val Gln Ser
20 25 30Met Thr Asn Thr Arg Thr Thr Asp Val Glu Ala Thr Val Asn Gln Ile
35 40 45Lys Ala Leu Glu Arg Val Gly Ala Asp Ile Val Arg Val Ser Val Pro
50 55 60Thr Met Asp Ala Ala Glu Ala Phe Lys Leu Ile Lys Gln Gln Val Asn 65 70 75 80Val Pro Leu Val Ala Asp Ile His Phe Asp Tyr Arg Ile Ala Leu Lys
85 90 95Val Ala Glu Tyr Gly Val Asp Cys Leu Arg Ile Asn Pro Gly Asn Ile
100 105 110Gly Asn Glu Glu Arg Ile Arg Met Val Val Asp Cys Ala Arg Asp Lys
115 120 125Asn Ile Pro Ile Arg Ile Gly Val Asn Ala Gly Ser Leu Glu Lys Asp
130 135 140Leu Gln Glu Lys Tyr Gly Glu Pro Thr Pro Gln Ala Leu Leu Glu Ser145 150 155 160Ala Met Arg His Val Asp His Leu Asp Arg Leu Asn Phe Asp Gln Phe
165 170 175Lys Val Ser Val Lys Ala Ser Asp Val Phe Leu Ala Val Glu Ser Tyr
180 185 190Arg Leu Leu Ala Lys Gln Ile Asp Gln Pro Leu His Leu Gly Ile Thr
195 200 205Glu Ala Gly Gly Ala Arg Ser Gly Ala Val Lys Ser Ala Ile Gly Leu
210 215 220Gly Leu Leu Leu Ser Glu Gly Ile Gly Asp Thr Leu Arg Val Ser Leu225 230 235 240Ala Ala Asp Pro Val Glu Glu Ile Lys Val Gly Phe Asp Ile Leu Lys
245 250 255Ser Leu Arg Ile Arg Ser Arg Gly Ile Asn Phe Ile Ala Cys Pro Thr
260 265 270Cys Ser Arg Gln Glu Phe Asp Val Ile Gly Thr Val Asn Ala Leu Glu
275 280 285Gln Arg Leu Glu Asp Ile Ile Thr Pro Met Asp Val Ser Ile Ile Gly
290 295 300Cys Val Val Asn Gly Pro Gly Glu Ala Leu Val Ser Thr Leu Gly Val305 310 315 320Thr Gly Gly Asn Lys Lys Ser Gly Leu Tyr Glu Asp Gly Val Arg Lys
325 330 335Asp Arg Leu Asp Asn Asn Asp Met Ile Asp Gln Leu Glu Ala Arg Ile
340 345 350Arg Ala Lys Ala Ser Gln Leu Asp Glu Ala Arg Arg Ile Asp Val Gln
355 360 365Gln Val Glu Lys
370<210>3<211>2109<212>DNA<213>Plasmodium falciparum<220><221>CDS<222>(70)..(2109)<400>3cagcctataa atattattat ttattattat tttttttttt ttttttcata atgcctgaat 60aaccacaaa atg agt tat ata aaa aga ctg att ctt ttt atg tta ctg ttt 111
Met Ser Tyr Ile Lys Arg Leu Ile Leu Phe Met Leu Leu Phe
1 5 10tat tct cat gta aaa att aaa aaa tta ttt att aaa att tct aat gta 159Tyr Ser His Val Lys Ile Lys Lys Leu Phe Ile Lys Ile Ser Asn Val 15 20 25 30aac ata ttt ttt gca gaa gca aag aaa aat gga aaa aag gaa ttc ttt 207Asn Ile Phe Phe Ala Glu Ala Lys Lys Asn Gly Lys Lys Glu Phe Phe
35 40 45ctt ttt tta cta aat ata aaa aaa aat agc caa cag aaa aaa act tat 255Leu Phe Leu Leu Asn Ile Lys Lys Asn Ser Gln Gln Lys Lys Thr Tyr
50 55 60cat att acc aaa agg aat acc ata aat aaa agt gat ttt tta tat tct 303His Ile Thr Lys Arg Asn Thr Ile Asn Lys Ser Asp Phe Leu Tyr Ser
65 70 75tta cta aat gaa gaa ggg aat tct tca aaa aag gaa tat aaa aat tta 351Leu Leu Asn Glu Glu Gly Asn Ser Ser Lys Lys Glu Tyr Lys Asn Leu
80 85 90aaa gat gaa gaa aaa tat aat atc ata caa aat ata aaa aaa tat tgt 399Lys Asp Glu Glu Lys Tyr Asn Ile Ile Gln Asn Ile Lys Lys Tyr Cys 95 100 105 110gaa tgt act aaa aaa tat aaa agg ctc cca aca cga gaa gta gtt att 447Glu Cys Thr Lys Lys Tyr Lys Arg Leu Pro Thr Arg Glu Val Val Ile
115 120 125gga aat gtt aaa att gga gga aat aat aaa ata gct att caa act atg 495Gly Asn Val Lys Ile Gly Gly Asn Asn Lys Ile Ala Ile Gln Thr Met
130 135 140gct agc tgt gat aca aga aat gta gaa gaa tgt gta tat caa att aga 543Ala Ser Cys Asp Thr Arg Asn Val Glu Glu Cys Val Tyr Gln Ile Arg
145 150 155aaa tgt aaa gat ttg ggt gct gac att gta agg ttg act gtt caa gga 591Lys Cys Lys Asp Leu Gly Ala Asp Ile Val Arg Leu Thr Val Gln Gly
160 165 170gtt caa gaa gca caa gct agt tat cat att aaa gaa aaa tta tta tct 639Val Gln Glu Ala Gln Ala Ser Tyr His Ile Lys Glu Lys Leu Leu Ser175 180 185 190gaa aat gta aat atc cca tta gta aca gat att cat ttt aat cct aaa 687Glu Asn Val Asn Ile Pro Leu Val Thr Asp Ile His Phe Asn Pro Lys
195 200 205ata gct tta atg gca gct gat gtg ttt gaa aaa att cga gtg aat cca 735Ile Ala Leu Met Ala Ala Asp Val Phe Glu Lys Ile Arg Val Asn Pro
210 215 220gga aat tat gtt gat gga aga aaa aaa tgg ata gat aaa gtt tat aaa 783Gly Asn Tyr Val Asp Gly Arg Lys Lys Trp Ile Asp Lys Val Tyr Lys
225 230 235act aaa gaa gaa ttt gat gaa ggg aaa tta ttt ata aaa gaa aaa ttt 831Thr Lys Glu Glu Phe Asp Glu Gly Lys Leu Phe Ile Lys Glu Lys Phe
240 245 250gta cca tta att gaa aaa tgt aaa aga tta aat aga gca ata aga att 879Val Pro Leu Ile Glu Lys Cys Lys Arg Leu Asn Arg Ala Ile Arg Ile255 260 265 270ggt aca aat cat gga ttc ctt tca tct cga gta tta tca tat tat gga 927Gly Thr Asn His Gly Phe Leu Ser Ser Arg Val Leu Ser Tyr Tyr Gly
275 280 285gat aca cea tta gca tta gta gaa agt gct atg aga ttt tct gat tta 975Asp Thr Pro Leu Ala Leu Val Glu Ser Ala Met Arg Phe Ser Asp Leu
290 295 300tgt aat gaa aac aat ttt aac aat ctt gtt ttt tct atg aaa gct tct 1023Cys Asn Glu Asn Asn Phe Asn Asn Leu Val Phe Ser Met Lys Ala Ser
305 310 315aat gct tat gtt atg ata caa tct tat aga tta tta gta tct aaa caa 1071Asn Ala Tyr Val Met Ile Gln Ser Tyr Arg Leu Leu Val Ser Lys Gln
320 325 330tat gaa aga aat atg atg ttc cct ata cat tta gga gtt aca gaa gca 1119Tyr Glu Arg Asn Met Met Phe Pro Ile His Leu Gly Val Thr Glu Ala335 340 345 350gga ttt ggt gat aat gga aga ata aaa tct tat tta ggt ata gga tct 1167Gly Phe Gly Asp Asn Gly Arg Ile Lys Ser Tyr Leu Gly Ile Gly Ser
355 360 365tta tta tat gat ggt ata gga gat acc att cgt ata tcc tta aca gaa 1215Leu Leu Tyr Asp Gly Ile Gly Asp Thr Ile Arg Ile Ser Leu Thr Glu
370 375 380gat cct tgg gaa gag tta act cct tgt aaa aaa tta gtt gaa aat tta 1263Asp Pro Trp Glu Glu Leu Thr Pro Cys Lys Lys Leu Val Glu Asn Leu
385 390 395aag aaa aga ata ttt tat aat gaa aat ttt aaa gaa gat aat gaa tta 1311Lys Lys Arg Ile Phe Tyr Asn Glu Asn Phe Lys Glu Asp Asn Glu Leu
400 405 410aaa aat aat gaa atg gat acc aaa aat cta tta aat ttt gaa gaa aat 1359Lys Asn Asn Glu Met Asp Thr Lys Asn Leu Leu Asn Phe Glu Glu Asn415 420 425 430tat cga aat ttt aat aat ata aaa aaa aga aat gta gaa aaa aat aat 1407Tyr Arg Asn Phe Asn Asn Ile Lys Lys Arg Asn Val Glu Lys Asn Asn
435 440 445aat gta tta cat gaa gag tgc act ata ggt aat gta gta acc ata aaa 1455Asn Val Leu His Glu Glu Cys Thr Ile Gly Asn Val Val Thr Ile Lys
450 455 460gag tta gaa gat tct ctg caa att ttt aaa gat tta aat tta gaa gta 1503Glu Leu Glu Asp Ser Leu Gln Ile Phe Lys Asp Leu Asn Leu Glu Val
465 470 475gat tca aat gga aat ttg aaa aag gga gcc aaa aca act gat atg gtt 1551Asp Ser Asn Gly Asn Leu Lys Lys Gly Ala Lys Thr Thr Asp Met Val
480 485 490att ata aat gat ttt cat aat ata aca aat tta gga aaa aaa act gtg 1599Ile Ile Asn Asp Phe His Asn Ile Thr Asn Leu Gly Lys Lys Thr Val495 500 505 510gat aaa tta atg caa gtg gga att aat ata gta gtt caa tat gaa cca 1647Asp Lys Leu Met Gln Val Gly Ile Asn Ile Val Val Gln Tyr Glu Pro
515 520 525cat aat ata gaa ttt ata gaa aaa atg gaa cca aat aat gat aat aat 1695His Asn Ile Glu Phe Ile Glu Lys Met Glu Pro Asn Asn Asp Asn Asn
530 535 540aat aat aat aat aat aat aat ata tta ttt tat gtg gat ata aaa aat 1743Asn Asn Asn Asn Asn Asn Asn Ile Leu Phe Tyr Val Asp Ile Lys Asn
545 550 555att atg aac agt tca gaa aaa aat att aaa tta agt aat tct aaa gga 1791Ile Met Asn Ser Ser Glu Lys Asn Ile Lys Leu Ser Asn Ser Lys Gly
560 565 570tat gga tta att tta aac gga aaa gaa gat ata caa acc ata aaa aaa 1839Tyr Gly Leu Ile Leu Asn Gly Lys Glu Asp Ile Gln Thr Ile Lys Lys575 580 585 590ata aaa gaa tta aat cgt cgt cct tta ttc att cta tta aaa tca gat 1887Ile Lys Glu Leu Asn Arg Arg Pro Leu Phe Ile Leu Leu Lys Ser Asp
595 600 605aac ata tat gaa cat gta tta ata acc aga aga att aat gaa ctt tta 1935Asn Ile Tyr Glu His Val Leu Ile Thr Arg Arg Ile Asn Glu Leu Leu
610 615 620caa tcc tta aat ata aat ata cct tat ata cat tat gtt gat att aat 1983Gln Ser Leu Asn Ile Asn Ile Pro Tyr Ile His Tyr Val Asp Ile Asn
625 630 635tca aat aat tat gat gat ata tta gtt aat tca aca tta tat gca gga 2031Ser Asn Asn Tyr Asp Asp Ile Leu Val Asn Ser Thr Leu Tyr Ala Gly
640 645 650agt tgt ttg atg gat tta atg ggg gat ggt ctt att gtt aac gta act 2079Ser Cys Leu Met Asp Leu Met Gly Asp Gly Leu Ile Val Asn Val Thr655 660 665 670aat gat gtt ctt aca aat aaa aaa ggg tag 2109Asn Asp Val Leu Thr Asn Lys Lys Gly
675 680<210>4<211>679<212>PRT<213>Plasmodium falciparum<400>4Met Ser Tyr Ile Lys Arg Leu Ile Leu Phe Met Leu Leu Phe Tyr Ser 1 5 10 15His Val Lys Ile Lys Lys Leu Phe Ile Lys Ile Ser Asn Val Asn Ile
20 25 30Phe Phe Ala Glu Ala Lys Lys Asn Gly Lys Lys Glu Phe Phe Leu Phe
35 40 45Leu Leu Asn Ile Lys Lys Asn Ser Gln Gln Lys Lys Thr Tyr His Ile
50 55 60Thr Lys Arg Asn Thr Ile Asn Lys Ser Asp Phe Leu Tyr Ser Leu Leu 65 70 75 80Asn Glu Glu Gly Asn Ser Ser Lys Lys Glu Tyr Lys Asn Leu Lys Asp
85 90 95Glu Glu Lys Tyr Asn Ile Ile Gln Asn Ile Lys Lys Tyr Cys Glu Cys
100 105 110Thr Lys Lys Tyr Lys Arg Leu Pro Thr Arg Glu Val Val Ile Gly Asn
115 120 125Val Lys Ile Gly Gly Asn Asn Lys Ile Ala Ile Gln Thr Met Ala Ser
130 135 140Cys Asp Thr Arg Asn Val Glu Glu Cys Val Tyr Gln Ile Arg Lys Cys145 150 155 160Lys Asp Leu Gly Ala Asp Ile Val Arg Leu Thr Val Gln Gly Val Gln
165 170 175Glu Ala Gln Ala Ser Tyr His Ile Lys Glu Lys Leu Leu Ser Glu Asn
180 185 190Val Asn Ile Pro Leu Val Thr Asp Ile His Phe Asn Pro Lys Ile Ala
195 200 205Leu Met Ala Ala Asp Val Phe Glu Lys Ile Arg Val Asn Pro Gly Asn
210 215 220Tyr Val Asp Gly Arg Lys Lys Trp Ile Asp Lys Val Tyr Lys Thr Lys225 230 235 240Glu Glu Phe Asp Glu Gly Lys Leu Phe Ile Lys Glu Lys Phe Val Pro
245 250 255Leu Ile Glu Lys Cys Lys Arg Leu Asn Arg Ala Ile Arg Ile Gly Thr
260 265 270Asn His Gly Phe Leu Ser Ser Arg Val Leu Ser Tyr Tyr Gly Asp Thr
275 280 285Pro Leu Ala Leu Val Glu Ser Ala Met Arg Phe Ser Asp Leu Cys Asn
290 295 300Glu Asn Asn Phe Asn Asn Leu Val Phe Ser Met Lys Ala Ser Asn Ala305 310 315 320Tyr Val Met Ile Gln Ser Tyr Arg Leu Leu Val Ser Lys Gln Tyr Glu
325 330 335Arg Asn Met Met Phe Pro Ile His Leu Gly Val Thr Glu Ala Gly Phe
340 345 350Gly Asp Asn Gly Arg Ile Lys Ser Tyr Leu Gly Ile Gly Ser Leu Leu
355 360 365Tyr Asp Gly Ile Gly Asp Thr Ile Arg Ile Ser Leu Thr Glu Asp Pro
370 375 380Trp Glu Glu Leu Thr Pro Cys Lys Lys Leu Val Glu Asn Leu Lys Lys385 390 395 400Arg Ile Phe Tyr Asn Glu Asn Phe Lys Glu Asp Asn Glu Leu Lys Asn
405 410 415Asn Glu Met Asp Thr Lys Asn Leu Leu Asn Phe Glu Glu Asn Tyr Arg
420 425 430Asn Phe Asn Asn Ile Lys Lys Arg Asn Val Glu Lys Asn Asn Asn Val
435 440 445Leu His Glu Glu Cys Thr Ile Gly Asn Val Val Thr Ile Lys Glu Leu
450 455 460Glu Asp Ser Leu Gln Ile Phe Lys Asp Leu Asn Leu Glu Val Asp Ser465 470 475 480Asn Gly Asn Leu Lys Lys Gly Ala Lys Thr Thr Asp Met Val Ile Ile
485 490 495Asn Asp Phe His Asn Ile Thr Asn Leu Gly Lys Lys Thr Val Asp Lys
500 505 510Leu Met Gln Val Gly Ile Asn Ile Val Val Gln Tyr Glu Pro His Asn
515 520 525Ile Glu Phe Ile Glu Lys Met Glu Pro Asn Asn Asp Asn Asn Asn Asn
530 535 540Asn Asn Asn Asn Asn Ile Leu Phe Tyr Val Asp Ile Lys Asn Ile Met545 550 555 560Asn Ser Ser Glu Lys Asn Ile Lys Leu Ser Asn Ser Lys Gly Tyr Gly
565 570 575Leu Ile Leu Asn Gly Lys Glu Asp Ile Gln Thr Ile Lys Lys Ile Lys
580 585 590Glu Leu Asn Arg Arg Pro Leu Phe Ile Leu Leu Lys Ser Asp Asn Ile
595 600 605Tyr Glu His Val Leu Ile Thr Arg Arg Ile Asn Glu Leu Leu Gln Ser
610 615 620Leu Asn Ile Asn Ile Pro Tyr Ile His Tyr Val Asp Ile Asn Ser Asn625 630 635 640Asn Tyr Asp Asp Ile Leu Val Asn Ser Thr Leu Tyr Ala Gly Ser Cys
645 650 655Leu Met Asp Leu Met Gly Asp Gly Leu Ile Val Asn Val Thr Asn Asp
660 665 670Val Leu Thr Asn Lys Lys Gly
675<210>5<211>1200<212>DNA<213>大肠杆菌<400>5aatggctatc agaccgcttt aatgtcgatg gcttcaccct gcatccgttt acgcagggta 60cgtttcgtac ggtcgataac atcgcccgcc aactgaccac aggcagcatc gatatcatca 120ccacgagttt tacgcacaat agtggtgaaa ccgtagctca tcagcacttt tgagaaacgg 180tcgatacggc tgttcgagct gcgtccatac ggcgcacccg ggaacgggtt ccacgggatc 240aggttgatct tacacggcgt atctttcagc agttccgcca gttggtgcgc gtgttcagtg 300ccgtcgttaa cgtggtcaag catcacgtat tcaatagtga ctcggccctg attggcgttg 360gatttctcca gataacggcg caccgcagca aggaacgttt cgatattgta ctttttgttg 420atcggcacaa tttcgtcacg aatttcgtcg ttcggcgcgt gcagggaaat tgccagtgca 480acgtcgatca tatcgcccag tttatccagc gccggaacta caccggaagt ggaaagcgtg 540acgcgacgtt tagacaggcc aaaaccgaaa tcatcaagca tgatttccat cgccggaacg 600acgttgttca ggttgagcag cggctcgccc atgcccatca tcactacgtt agtgatcgga 660cgctgaccgg tgacttttgc tgcgccgacg attttcgccg cacgccacac ctggccgata 720atttccgaca cccgcaggtt gcggttaaag ccctgctggg cggtggaaca gaatttacac 780tccagcgcac accccacctg cgaagagacg cagagcgtgg cacggtcgtc ttccgggata 840tacaccgttt cgacgcgctg atcgccaacg gcgatcgccc atttaatggt gccgtcagat 900gaacgctgtt cttcaaccac ttccggtgcg cggatttccg ccacctcttt cagtttgccg 960cgcaacactt tgttgatgtc ggtcatctca tcaaagttgt cgcagcaata gtgatacatc 1020cacttcatca cctgatcggc gcggaagggt ttttcaccta aatctttaaa aaactcccgc 1080atctgctgac ggttgagatc cagcaggttg atttttccat ctttcgtggt gacgttttca 1140ggtgtgacta attgttcaga catatgctat tccggcctcg ttattacacg ttatggcccc 1200<210>6<211>384<212>PRT<213>大肠杆菌<400>6Met Ser Glu Gln Leu Val Thr Pro Glu Asn Val Thr Thr Lys Asp Gly 1 5 10 15Lys Ile Asn Leu Leu Asp Leu Asn Arg Gln Gln Met Arg Glu Phe Phe
20 25 30Lys Asp Leu Gly Glu Lys Pro Phe Arg Ala Asp Gln Val Met Lys Trp
35 40 45Met Tyr His Tyr Cys Cys Asp Asn Phe Asp Glu Met Thr Asp Ile Asn
50 55 60Lys Val Leu Arg Gly Lys Leu Lys Glu Val Ala Glu Ile Arg Ala Pro 65 70 75 80Glu Val Val Glu Glu Gln Arg Ser Ser Asp Gly Thr Ile Lys Trp Ala
85 90 95Ile Ala Val Gly Asp Gln Arg Val Glu Thr Val Tyr Ile Pro Glu Asp
100 105 110Asp Arg Ala Thr Leu Cys Val Ser Ser Gln Val Gly Cys Ala Leu Glu
115 120 125Cys Lys Phe Cys Ser Thr Ala Gln Gln Gly Phe Asn Arg Asn Leu Arg
130 135 140Val Ser Glu Ile Ile Gly Gln Val Trp Arg Ala Ala Lys Ile Val Gly145 150 155 160Ala Ala Lys Val Thr Gly Gln Arg Pro Ile Thr Asn Val Val Met Met
165 170 175Gly Met Gly Glu Pro Leu Leu Asn Leu Asn Asn Val Val Pro Ala Met
180 185 190Glu Ile Met Leu Asp Asp Phe Gly Phe Gly Leu Ser Lys Arg Arg Val
195 200 205Thr Leu Ser Thr Ser Gly Val Val Pro Ala Leu Asp Lys Leu Gly Asp
210 215 220Met Ile Asp Val Ala Leu Ala Ile Ser Leu His Ala Pro Asn Asp Glu225 230 235 240Ile Arg Asp Glu Ile Val Pro Ile Asn Lys Lys Tyr Asn Ile Glu Thr
245 250 255Phe Leu Ala Ala Val Arg Arg Tyr Leu Glu Lys Ser Asn Ala Asn Gln
260 265 270Gly Arg Val Thr Ile Glu Tyr Val Met Leu Asp His Val Asn Asp Gly
275 280 285Thr Glu His Ala His Gln Leu Ala Glu Leu Leu Lys Asp Thr Pro Cys
290 295 300Lys Ile Asn Leu Ile Pro Trp Asn Pro Phe Pro Gly Ala Pro Tyr Gly305 310 315 320Arg Ser Ser Asn Ser Arg Ile Asp Arg Phe Ser Lys Val Leu Met Ser
325 330 335Tyr Gly Phe Thr Thr Ile Val Arg Lys Thr Arg Gly Asp Asp Ile Asp
340 345 350Ala Ala Cys Gly Gln Leu Ala Gly Asp Val Ile Asp Arg Thr Lys Arg
355 360 365Thr Leu Arg Lys Arg Met Gln Gly Glu Ala Ile Asp Ile Lys Ala Val
370 375 380<210>7<211>1320<212>DNA<213>Plasmodium falciparum<220><221>CDS<222>(163)..(1173)<400>7taaataaata aattataaat ctttcaagaa tatatttttt ataaaaacat aaaatataaa 60atatacatat atatatatat atatatttta tattactttt aaaattattt atttatacaa 120atggaaattt aatgtgaaga atagaaaaaa cattttgtca at atg gaa aag tca 174
Met Glu Lys Ser
1aaa agg tac ata age ctg att aag atg atg gaa agg aaa aaa ttt gag 222Lys Arg Tyr Ile Ser Leu Ile Lys Met Met Glu Arg Lys Lys Phe Glu 5 10 15 20aag tat aga tta aaa caa ata atg gat aat ata tat aaa gga aaa ata 270Lys Tyr Arg Leu Lys Gln Ile Met Asp Asn Ile Tyr Lys Gly Lys Ile
25 30 35att gaa ata aat aaa atg aaa aat att cca act gaa ata aga aga gaa 318Ile Glu Ile Asn Lys Met Lys Asn Ile Pro Thr Glu Ile Arg Arg Glu
40 45 50tta aaa aat ata ttt cat aat aat att tta agt ata aaa ccg atc aaa 366Leu Lys Asn Ile Phe His Asn Asn Ile Leu Ser Ile Lys Pro Ile Lys
55 60 65gaa tta aaa tat gat aga gca tat aaa gta tta ttt cag tgt aaa gat 414Glu Leu Lys Tyr Asp Arg Ala Tyr Lys Val Leu Phe Gln Cys Lys Asp
70 75 80aat gaa aag att gaa gca aca tca tta gat ttt ggt tcg cat aaa tct 462Asn Glu Lys Ile Glu Ala Thr Ser Leu Asp Phe Gly Ser His Lys Ser 85 90 95 100tta tgt ata tct agc caa ata ggt tgt tct ttt gga tgt aag ttt tgt 510Leu Cys Ile Ser Ser Gln Ile Gly Cys Ser Phe Gly Cys Lys Phe Cys
105 110 115gct act ggt caa att ggt ata aaa aga caa tta gat ata gat gaa ata 558Ala Thr Gly Gln Ile Gly Ile Lys Arg Gln Leu Asp Ile Asp Glu Ile
120 125 130act gat caa ctt tta tat ttt caa tca aaa gga gtt gat ata aaa aat 606Thr Asp Gln Leu Leu Tyr Phe Gln Ser Lys Gly Val Asp Ile Lys Asn
135 140 145ata tct ttt atg ggt atg gga gaa cct tta gct aat cca tat gtt ttt 654Ile Ser Phe Met Gly Met Gly Glu Pro Leu Ala Asn Pro Tyr Val Phe
150 155 160gat tct ata caa ttt ttt aat gat aat aat tta ttt tct ata tct aat 702Asp Ser Ile Gln Phe Phe Asn Asp Asn Asn Leu Phe Ser Ile Ser Asn165 170 175 180aga cgt att aat ata tct act gtt ggt ctt tta cca gga att aaa aaa 750Arg Arg Ile Asn Ile Ser Thr Val Gly Leu Leu Pro Gly Ile Lys Lys
185 190 195tta aat aac atc ttt cct caa gtt aat tta gct ttc tca tta cat tct 798Leu Asn Asn Ile Phe Pro Gln Val Asn Leu Ala Phe Ser Leu His Ser
200 205 210cca ttt act gaa gaa agg gat caa ctt gta cca att aat aaa ttg ttt 846Pro Phe Thr Glu Glu Arg Asp Gln Leu Val Pro Ile Asn Lys Leu Phe
215 220 225ccg ttt aat gaa gtt ttt gat tta tta gat gaa aga ata gca aaa act 894Pro Phe Asn Glu Val Phe Asp Leu Leu Asp Glu Arg Ile Ala Lys Thr
230 235 240ggt aga aga gtt tgg ata agt tat att tta att aaa aat ctt aat gac 942Gly Arg Arg Val Trp Ile Ser Tyr Ile Leu Ile Lys Asn Leu Asn Asp245 250 255 260tcc aaa gat cat gca gaa gct ttg tct gat cat ata tgt aaa aga cca 990Ser Lys Asp His Ala Glu Ala Leu Ser Asp His Ile Cys Lys Arg Pro
265 270 275aat aac ata aga tac tta tat aat gta tgt tta ata cct tat aat aaa 1038Asn Asn Ile Arg Tyr Leu Tyr Asn Val Cys Leu Ile Pro Tyr Asn Lys
280 285 290ggt aat aga att tat aat ata tca ttt gaa tat ata tat ata tat ata 1086Gly Asn Arg Ile Tyr Asn Ile Ser Phe Glu Tyr Ile Tyr Ile Tyr Ile
295 300 305tat tta cta ata ata aaa aaa aag ata tta tgt aaa tat att atg ttt 1134Tyr Leu Leu Ile Ile Lys Lys Lys Ile Leu Cys Lys Tyr Ile Met Phe
310 315 320cac aca tta tat aaa tat ata ggc ata gag gac atg tta taaaaaagtg 1183His Thr Leu Tyr Lys Tyr Ile Gly Ile Glu Asp Met Leu325 330 335caacatatat atatatatat atatatatat atatatatat acattttttt tatatttata 1243ttatcttttt aatacattta ttccattaca ttgcagccaa aaatgttgac gaaaattttc 1303atcgtttgga cgatgct 1320<210>8<211>337<212>PRT<213>Plasmodium falciparum<400>8Met Glu Lys Ser Lys Arg Tyr Ile Ser Leu Ile Lys Met Met Glu Arg 1 5 10 15Lys Lys Phe Glu Lys Tyr Arg Leu Lys Gln Ile Met Asp Asn Ile Tyr
20 25 30Lys Gly Lys Ile Ile Glu Ile Asn Lys Met Lys Asn Ile Pro Thr Glu
35 40 45Ile Arg Arg Glu Leu Lys Asn Ile Phe His Asn Asn Ile Leu Ser Ile
50 55 60Lys Pro Ile Lys Glu Leu Lys Tyr Asp Arg Ala Tyr Lys Val Leu Phe 65 70 75 80Gln Cys Lys Asp Asn Glu Lys Ile Glu Ala Thr Ser Leu Asp Phe Gly
85 90 95Ser His Lys Ser Leu Cys Ile Ser Ser Gln Ile Gly Cys Ser Phe Gly
100 105 110Cys Lys Phe Cys Ala Thr Gly Gln Ile Gly Ile Lys Arg Gln Leu Asp
115 120 125Ile Asp Glu Ile Thr Asp Gln Leu Leu Tyr Phe Gln Ser Lys Gly Val
130 135 140Asp Ile Lys Asn Ile Ser Phe Met Gly Met Gly Glu Pro Leu Ala Asn145 150 155 160Pro Tyr Val Phe Asp Ser Ile Gln Phe Phe Asn Asp Asn Asn Leu Phe
165 170 175Ser Ile Ser Asn Arg Arg Ile Asn Ile Ser Thr Val Gly Leu Leu Pro
180 185 190Gly Ile Lys Lys Leu Asn Asn Ile Phe Pro Gln Val Asn Leu Ala Phe
195 200 205Ser Leu His Ser Pro Phe Thr Glu Glu Arg Asp Gln Leu Val Pro Ile
210 215 220Asn Lys Leu Phe Pro Phe Asn Glu Val Phe Asp Leu Leu Asp Glu Arg225 230 235 240Ile Ala Lys Thr Gly Arg Arg Val Trp Ile Ser Tyr Ile Leu Ile Lys
245 250 255Asn Leu Asn Asp Ser Lys Asp His Ala Glu Ala Leu Ser Asp His Ile
260 265 270Cys Lys Arg Pro Asn Asn Ile Arg Tyr Leu Tyr Asn Val Cys Leu Ile
275 280 285Pro Tyr Asn Lys Gly Asn Arg Ile Tyr Asn Ile Ser Phe Glu Tyr Ile
290 295 300Tyr Ile Tyr Ile Tyr Leu Leu Ile Ile Lys Lys Lys Ile Leu Cys Lys305 310 315 320Tyr Ile Met Phe His Thr Leu Tyr Lys Tyr Ile Gly Ile Glu Asp Met
325 330 335Leu